Apr. 2025, 市村清新技術財団, 市村学術賞（功績賞）　　　　　　　　　　　　　　　
遺伝子・核酸送達法が拓く革新的ナノ医療
原島秀吉

Apr. 2024, 秋山記念生命科学振興財団, 秋山財団賞　　　　　　　　　　　　　　　
細胞内動態制御から革新的医薬品の創出へ
原島秀吉

Mar. 2023, Pharmaceutical Society of Japan, Awad of Phamaceutical Society of Japan　　　　　　　　　　　　　　　
細胞内動態制御に基づいた遺伝子・核酸送達システムの創製とナノ医療への展開
Hideyoshi Harashima

Sep. 2021, International Federation of Pharmaceutics, Høst Madsen Medal　　　　　　　　　　　　　　　
Multifunctional Envelope-type Nano Device: From controlled intracellular trafficking to clinical application for Nanomedicine
Hideyoshi Harashima

Feb. 2019, Hokkaido University, Presidential Award　　　　　　　　　　　　　　　
Hideyoshi Harashima

Nov. 2016, Song Eum Academy Foundation, 19th SONG EUM Med-Pharm Award　　　　　　　　　　　　　　　
Hideyoshi Harashima

May 2016, 公益社団法人日本薬剤学会, 学会賞　　　　　　　　　　　　　　　
多機能性エンベロープ型ナノ構造体の創製とナノ医療への展開
原島秀吉

Feb. 2016, 北海道大学, 総長賞（優秀賞）　　　　　　　　　　　　　　　
原島秀吉

Feb. 2016, 一般財団法人油脂工業会館, 優秀論文表彰（優秀賞）　　　　　　　　　　　　　　　
秋田英万;原島秀吉

Feb. 2015, 北海道大学, 総長賞（優秀賞）　　　　　　　　　　　　　　　
原島秀吉

Jul. 2013, 国際コントロールドリリース学会, Fellow　　　　　　　　　　　　　　　
原島秀吉

Oct. 2011, 米国薬学会, AAPS Journal Manuscript Award　　　　　　　　　　　　　　　
A. El-Sayed;Shiro Futaki;Hideyoshi Harashima

Aug. 2010, FIP, Distinguished Science Award　　　　　　　　　　　　　　　
Hideyoshi Harashima

Jun. 2007, 日本DDS学会, 永井賞　　　　　　　　　　　　　　　
多機能性エンベロープ型ナノ構造体による人工遺伝子デリバリーシステムの創製
原島秀吉

Mar. 2007, 公益社団法人日本薬学会, 学術振興賞　　　　　　　　　　　　　　　
多機能性エンベロープ型人工遺伝子デリバリーシステムの創製
原島秀吉

Nov. 1994, 日本薬物動態学会, 奨励賞　　　　　　　　　　　　　　　
薬物速度論に基づいたリポソームによる薬物送達システムの構築
原島秀吉

Harnessing the composition of lipid nanoparticles to selectively deliver mRNA to splenic immune cells for anticancer vaccination
Mahmoud A. Younis, Yusuke Sato, Yaser H. A. Elewa, Hideyoshi Harashima
Drug Delivery and Translational Research, Springer Science and Business Media LLC, 07 Mar. 2025
Scientific journal

Targeting Tumor Endothelial Cells by EGCG Using Specific Liposome Delivery System Inhibits Vascular Inflammation and Thrombosis.
Zi Jia, Nako Maishi, Hideki Takekawa, Aya Yanagawa Matsuda, Taisei Nakade, Takashi Nakamura, Hideyoshi Harashima, Yasuhiro Hida, Kyoko Hida
Cancer medicine, 13, 23, e70462, Dec. 2024, [International Magazine]
English, Scientific journal, BACKGROUND: Inflammation is one of the hallmarks of cancer and is associated with tumor growth. Tumor endothelial cells (TECs) demonstrate inflamed phenotypes. Endothelial inflammation initiates thrombus formation, which is the second cause of cancer-related deaths. Epigallocatechin-3-O-gallate (EGCG), a natural compound in green tea, has demonstrated an anti-inflammatory effect. However, the tumor progression inhibition effect of EGCG by targeting TEC inflammation remains unclear. This study addresses the anti-tumor effect of EGCG, especially its anti-inflammatory role in TECs. METHODS: In vitro, the effect of EGCG on TECs were studied using real-time quantitative PCR and immunofluoresence to analyza gene and protein expression. In vivo, a cyclic RGD liposome delivery system (MEND) was employed to efficiently deliver EGCG to TECs in tumor-bearing mice. RESULTS: In vitro, EGCG significantly reduces inflammatory cytokine expression, including tumor necrosis factor-α, interleukin-6, IL-8, and IL-1β through NF-κB signaling inhibition. Additionally, von Willebrand factor reduction in TECs, which is involved in platelet adhesion and thrombosis formation, was analyzed. Our results revealed that EGCG-MEND significantly inhibited TEC inflammation and thrombus formation in tumors. Additionally, EGCG-MEND improved tumor immunity by reducing programmed death-ligand 1 expression and promoting high endothelial venule formation by recruiting CD8+ T cells. CONCLUSION: Our results indicate the anti-tumor potential of EGCG-MEND in normalizing the inflammatory immune microenvironment and inhibiting thrombosis by targeting TEC.

Engineering branched ionizable lipid for hepatic delivery of clustered regularly interspaced short palindromic repeat-Cas9 ribonucleoproteins.
Haruno Onuma, Rina Shimizu, Yuichi Suzuki, Mina Sato, Hideyoshi Harashima, Yusuke Sato
iScience, 27, 10, 110928, 110928, 18 Oct. 2024, [International Magazine]
English, Scientific journal, The delivery of the CRISPR/Cas ribonucleoprotein (RNP) has received attention for clinical applications owing to its high efficiency with few off-target effects. Lipid nanoparticles (LNPs) are potential non-viral vectors for the delivery of RNPs. Herein, we report the engineering of a branched scaffold structure of ionizable lipids for the hepatic delivery of RNPs. Both the total carbon number and branching position were critical for the functional delivery of RNPs. The optimal ionizable lipid exhibited a more than 98% reduction in transthyretin protein after a single dose with no obvious signs of toxicity. The mechanistic study has revealed that optimal LNPs have a unique "flower-like structure" that depends on both the lipid structure and the payload and that these LNPs accumulate in hepatocytes in an apolipoprotein E-independent manner. These results represent a major step toward the realization of in vivo genome editing therapy via RNP delivery using chemically synthesizable LNP formulations.

Impact of in vivo fate of STING agonist-loaded lipid nanoparticles on antitumor immunity.
Rikito Endo, Tomoki Ueda, Takumi Nagaoki, Natsumi Shima, Yusuke Sato, Hideyoshi Harashima, Takashi Nakamura
Journal of controlled release : official journal of the Controlled Release Society, 372, 609, 618, Aug. 2024, [International Magazine]
English, Scientific journal, Therapeutically manipulating the stimulator of interferon genes (STING) pathway has promising potential for enhancing antitumor immunity. Agonists of this pathway (STING agonists) are being evaluated in clinical trials. Loading the STING agonists into lipid nanoparticles (LNPs) increases their safety and efficacy. We previously developed STING agonists loaded LNPs consisting of the ionizable lipid YSK12-C4 (YSK12-LNPs), which showed significant antitumor effects. However, it is largely unclear how the in vivo fate of STING agonists loaded LNPs affects the antitumor immune responses. In this study, we compared the YSK12-LNPs with LNPs composed of DLin-MC3-DMA (MC3-LNPs) showing different in vivo fates. Biodistribution and flow cytometry analyses of mouse tissues revealed that the MC3-LNPs delivered higher amounts of STING agonists to the liver than the YSK12-LNPs. Additionally, significantly more liver leukocytes internalized the MC3-LNPs than the YSK12-LNPs. In contrast, the YSK12-LNPs delivered higher amounts of STING agonists to the liver leukocytes than the MC3-LNPs, leading to the effective induction of innate immunity and inflammation in the tumors. However, the antitumor effects in the B16-F10 lung metastasis and CT26 tumor models were comparable. Interestingly, flow cytometry analyses suggested that the YSK12-LNPs were more likely to activate natural killer cells and M1 macrophages, while the MC3-LNPs were more likely to activate CD8+ T cells. Our data suggest that different antitumor immune response mechanisms may operate depending on the characteristics and distribution of the LNPs.

The impact of, and expectations for, lipid nanoparticle technology: From cellular targeting to organelle targeting.
Yusuke Sato, Takashi Nakamura, Yuma Yamada, Hideyoshi Harashima
Journal of controlled release : official journal of the Controlled Release Society, 370, 516, 527, Jun. 2024, [International Magazine]
English, Scientific journal, The success of mRNA vaccines against COVID-19 has enhanced the potential of lipid nanoparticles (LNPs) as a system for the delivery of mRNA. In this review, we describe our progress using a lipid library to engineer ionizable lipids and promote LNP technology from the viewpoints of safety, controlled biodistribution, and mRNA vaccines. These advancements in LNP technology are applied to cancer immunology, and a potential nano-DDS is constructed to evaluate immune status that is associated with a cancer-immunity cycle that includes the sub-cycles in tumor microenvironments. We also discuss the importance of the delivery of antigens and adjuvants in enhancing the cancer-immunity cycle. Recent progress in NK cell targeting in cancer immunotherapy is also introduced. Finally, the impact of next-generation DDS technology is explained using the MITO-Porter membrane fusion-based delivery system for the organelle targeting of the mitochondria. We introduce a successful example of the MITO-Porter used in a cell therapeutic strategy to treat cardiomyopathy.

Duchenne型筋ジストロフィーモデルラット細胞を用いたミトコンドリア呼吸能活性化による治療介入検討　　　　　　　　　　　　　　　
佐藤 逸美, 日比野 光恵, 佐々木 大輔, 武田 充人, 原島 秀吉, 山田 勇磨
脳と発達, 56, Suppl., S237, S237, (一社)日本小児神経学会, May 2024
Japanese

Investigation of the Nanoparticulation Method and Cell-Killing Effect following the Mitochondrial Delivery of Hydrophobic Porphyrin-Based Photosensitizers
Rina Naganawa, Hanjun Zhao, Yuta Takano, Masatoshi Maeki, Manabu Tokeshi, Hideyoshi Harashima, Yuma Yamada
International Journal of Molecular Sciences, 12 Apr. 2024, [Peer-reviewed]
Scientific journal

Vaccination with a combination of STING agonist-loaded lipid nanoparticles and CpG-ODNs protects against lung metastasis via the induction of CD11bhighCD27low memory-like NK cells.
Alaa M Khalifa, Takashi Nakamura, Yusuke Sato, Hideyoshi Harashima
Experimental hematology & oncology, 13, 1, 36, 36, 29 Mar. 2024, [International Magazine]
English, Scientific journal, BACKGROUND: Natural killer (NK) cells are effective in attacking tumor cells that escape T cell attack. Memory NK cells are believed to function as potent effector cells in cancer immunotherapy. However, knowledge of their induction, identification, and potential in vivo is limited. Herein, we report on the induction and identification of memory-like NK cells via the action of a combination of a stimulator of interferon genes (STING) agonist loaded into lipid nanoparticles (STING-LNPs) and cytosine-phosphorothioate-guanine oligodeoxynucleotides (CpG-ODNs), and the potential of the inducted memory-like NK cells to prevent melanoma lung metastasis. METHODS: The antitumor effects of either the STING-LNPs, CpG-ODNs, or the combination therapy were evaluated using a B16-F10 lung metastasis model. The effect of the combined treatment was evaluated by measuring cytokine production. The induction of memory-like NK cells was demonstrated via flow cytometry and confirmed through their preventative effect. RESULTS: The combination of STING-LNPs and CpG-ODNs tended to enhance the production of interleukin 12 (IL-12) and IL-18, and exerted a therapeutic effect against B16-F10 lung metastasis. The combination therapy increased the population of CD11bhighCD27low NK cells. Although monotherapies failed to show preventative effects, the combination therapy induced a surprisingly strong preventative effect, which indicates that CD11bhighCD27low cells could be a phenotype of memory-like NK cells. CONCLUSION: As far as could be ascertained, this is the first report of the in vivo induction, identification, and confirmation of a phenotype of the memory-like NK cells through a prophylactic effect via the use of an immunotherapeutic drug. Our findings provide novel insights into the in vivo induction of CD11bhighCD27low memory-like NK cells thus paving the way for the development of efficient immunotherapies.

Activation of Mitochondrial Oxygen Consumption Rate by Delivering Coenzyme Q10 to Mitochondria of Rat Skeletal Muscle Cell (L6)
Itsumi Sato, Mitsue Hibino, Atsuhito Takeda, Hideyoshi Harashima, Yuma Yamada
Journal of Pharmaceutical Sciences, Elsevier BV, Feb. 2024
Scientific journal

Human cardiosphere-derived cells with activated mitochondria for better myocardial regenerative therapy.
Masahiro Shiraishi, Daisuke Sasaki, Mitsue Hibino, Atsuhito Takeda, Hideyoshi Harashima, Yuma Yamada
Journal of controlled release : official journal of the Controlled Release Society, 29 Jan. 2024, [International Magazine]
English, Scientific journal, Cell transplantation is a promising therapeutic strategy for myocardial regeneration therapy. To improve therapeutic effects, we developed a culture medium additive that enhances the mitochondrial function of cardiomyocytes for transplantation. A mitochondrial targeting drug delivery system (MITO-Porter system) was used to deliver mitochondrial activation molecules to mouse-derived cardiac progenitor cells. In this study, we investigated whether the mitochondrial function of human-derived myocardial precursor cells could be enhanced using MITO-Porter. Human cardiosphere-derived cells (CDCs) were isolated from myocardium which was excised during surgery for congenital heart disease. MITO-Porter was added to the cell culture medium to generate mitochondrial activated CDCs (human MITO cells). The human MITO cells were transplanted into myocardial ischemia-reperfusion rat model rat, and the effect was investigated. The transplanted human MITO cells improved the cardiac function and suppressed myocardial fibrosis compared to conventional cell transplantation methods. These effects were observed not only with myocardial administration but also by intravenous administration of human MITO cells. This study is the first study that assessed whether the mitochondrial delivery of functional compounds improved the outcome of human-derived myocardial cell transplantation therapy.

Development of Polymer-Lipid Hybrid Nanoparticles for Large-Sized Plasmid DNA Transfection.
Masatoshi Maeki, Shuya Uno, Kaisei Sugiura, Yusuke Sato, Yoichiro Fujioka, Akihiko Ishida, Yusuke Ohba, Hideyoshi Harashima, Manabu Tokeshi
ACS applied materials & interfaces, 16, 2, 2110, 2119, 17 Jan. 2024, [International Magazine]
English, Scientific journal, RNA and DNA delivery technologies using lipid nanoparticles (LNPs) have advanced significantly, as demonstrated by their successful application in mRNA vaccines. To date, commercially available RNA therapeutics include Onpattro, a 21 bp siRNA, and mRNA vaccines comprising 4300 nucleotides for COVID-19. However, a significant challenge remains in achieving efficient transfection, as the size of the delivered RNA and DNA increases. In contrast to RNA transfection, plasmid DNA (pDNA) transfection requires multiple steps, including cellular uptake, endosomal escape, nuclear translocation, transcription, and translation. The low transfection efficiency of large pDNA is a critical limitation in the development of artificial cells and their cellular functionalization. Here, we introduce polymer-lipid hybrid nanoparticles designed for efficient, large-sized pDNA transfection. We demonstrated that LNPs loaded with positively charged pDNA-polycation core nanoparticles exhibited a 4-fold increase in transfection efficiency for 15 kbp pDNA compared with conventional LNPs, which encapsulate a negatively charged pDNA-polycation core. Based on assessments of the size and internal structure of the polymer-lipid nanoparticles as well as hemolysis and cellular uptake analysis, we propose a strategy to enhance large-sized pDNA transfection using LNPs. This approach holds promise for accelerating the in vivo delivery of large-sized pDNA and advancing the development of artificial cells.

Development of Liposomes That Target Axon Terminals Encapsulating Berberine in Cultured Primary Neurons
Ikuma Hori, Hideyoshi Harashima, Yuma Yamada
Pharmaceutics, 16, 1, 49, 49, MDPI AG, 28 Dec. 2023
Scientific journal, Most of the energy in neurons is produced in mitochondria. Mitochondria generate the ATP that is essential for neuronal growth, function, and regeneration. Mitochondrial axonal transport plays a crucial role in maintaining neuronal homeostasis and biological activity. Decreased mitochondrial axonal transport at axon terminals, where the metabolism of substances is likely to be delayed, may contribute to neurological dysfunction. Therefore, regulation of mitochondrial dynamics at axon terminals has attracted considerable interest as a strategy to modulate neuronal function. Nanoparticles may be useful in controlling local mitochondrial dynamics. Nevertheless, there are few reports on the influence of drug delivery that nanoparticles impart on the mitochondrial dynamics in neurons. This paper reports the results of a study using liposomes (LPs) to examine local drug delivery and pharmacological actions on neurons. We tested berberine (BBR), which is an activator of AMP-activated protein kinase (AMPK), to examine the utility of this drug as a cellular energy sensor. Axon terminals targeting LPs were prepared. The amount of axon terminals targeting LPs was increased compared with treatment using cationic LPs. Moreover, axon terminal-targeting LPs increased anterograde transport by about 40% compared with that of either naked BBR or cationic LPs and suppressed axonal retraction. Our findings suggest that local drug delivery to neurons is important for enhancing pharmacological activity in axon terminals.

Cancer Immunotherapy with Lipid Nanoparticles Loaded with a Stimulator of Interferon Genes Agonist against Renal Tumor Lung Metastasis.
Takashi Nakamura, Shun Sasaki, Yusuke Sato, Hideyoshi Harashima
Pharmaceutics, 16, 1, 26 Dec. 2023, [International Magazine]
English, Scientific journal, Metastatic renal cell carcinoma (RCC) has a poor prognosis, and the major organ of metastasis is the lung. Immunotherapy with immune checkpoint inhibitors (ICIs) is the first-line therapy, but the response rates are low. Thus, the development of a more effective immunotherapy against metastatic RCC would be highly desirable. We previously demonstrated how a stimulator of an interferon gene (STING) agonist-loaded lipid nanoparticles (STING-LNPs) significantly activates natural killer (NK) cells and induces an antitumor effect against cases of melanoma lung metastasis that have shown ICI resistance. In this study, we evaluated the potential of using STING-LNPs in the treatment of lung metastatic RCC (Renca). An intravenous injection of STING-LNPs drastically decreased the amount of Renca tumor colonies. In contrast, monotherapies using ICIs showed no antitumor effect, and even a combination of ICI and STING-LNP therapies failed to enhance the antitumor effects. The main effector cells would be NK cells, and the activation of NK cells by the STING-LNPs may avoid the increased expression of immune checkpoint molecules. These findings provide useful insights into the development of an effective immunotherapy against metastatic RCC.

π-extended porphyrin-based near-infrared photosensitizers for mitochondria-targeted photodynamic therapy
Hanjun Zhao, Rina Naganawa, Yuma Yamada, Yasuko Osakada, Mamoru Fujitsuka, Hideyuki Mitomo, Yukiko Miyatake, Hideyoshi Harashima, Vasudevanpillai Biju, Yuta Takano
Journal of Photochemistry and Photobiology A: Chemistry, 115397, 115397, Elsevier BV, Dec. 2023
Scientific journal

Reprogramming activated hepatic stellate cells by siRNA-loaded nanocarriers reverses liver fibrosis in mice
Mahmoud A. Younis, Yusuke Sato, Yaser H.A. Elewa, Hideyoshi Harashima
Journal of Controlled Release, 361, 592, 603, Sep. 2023
Scientific journal

Harnessing Topology and Stereochemistry of Glycidylamine‐Derived Lipid Nanoparticles for in Vivo mRNA Delivery to Immune Cells in Spleen and Their Application for Cancer Vaccination
Mahmoud M. Abd Elwakil, Ryota Suzuki, Alaa M. Khalifa, Rania M. Elshami, Takuya Isono, Yaser H.A. Elewa, Yusuke Sato, Takashi Nakamura, Toshifumi Satoh, Hideyoshi Harashima
Advanced Functional Materials, Wiley, 26 Jul. 2023
Scientific journal, Abstract

mRNA lipid nanoparticles (LNPs) have reached an inflection point and are now paving the way for a new wave of precision therapies. The design of nonhepatocyte RNA delivery systems without targeting ligands, however, remains a challenge. It is reported that the development of ligand‐free glycidylamine (GA) derivatives containing LNPs (GA‐LNPs) that preferentially deliver mRNA to immune cells in the spleen. Notably, it is demonstrated that the stereochemistry of GA‐lipids has a significant impact on their self‐assembly and in vitro and in vivo RNA delivery efficiency and tropism. This impact is dependent on the monomeric structure of GA and number of stereogenic centers. Furthermore, the nonlinear topology of GA lipid derivatives induced a sevenfold improvement in mRNA delivery efficiency. The top‐performing estriol‐GA05‐30 LNPs elicited strong antitumor activity in a therapeutic and prophylactic cancer model and are well tolerated in mice. These results highlight the significance of the chemistry of ionizable lipids for extrahepatic RNA delivery and indicated a promising direction for the development of next‐generation mRNA immunotherapies.

骨格筋細胞ミトコンドリアに対する薬物送達および機能活性化についての検討　　　　　　　　　　　　　　　
佐藤 逸美, 日比野 光恵, 佐々木 大輔, 武田 充人, 原島 秀吉, 山田 勇磨
日本DDS学会学術集会プログラム予稿集, 39回, 144, 144, 日本DDS学会, Jul. 2023
Japanese

ミトコンドリア活性化幹細胞(MITO cell)を用いた心筋虚血再灌流モデルマウスにおける細胞移植治療の検証　　　　　　　　　　　　　　　
佐々木 大輔, 武田 充人, 原島 秀吉, 山田 勇磨
日本DDS学会学術集会プログラム予稿集, 39回, 176, 176, 日本DDS学会, Jul. 2023
Japanese

オミックス解析がもたらす小児心血管疾患研究の新たな展開 メタボローム解析を用いたミトコンドリア活性化ヒト心筋前駆細胞移植法の効果検証　　　　　　　　　　　　　　　
白石 真大, 佐々木 大輔, 武田 充人, 原島 秀吉, 山田 勇磨
日本小児循環器学会総会・学術集会抄録集, 59回, [III, 04], (NPO)日本小児循環器学会, Jul. 2023
Japanese

Development of light-induced disruptive liposomes (LiDL) as a photoswitchable carrier for intracellular substance delivery.
Taichi Tsuneishi, Keiichi Kojima, Fumika Kubota, Hideyoshi Harashima, Yuma Yamada, Yuki Sudo
Chemical communications (Cambridge, England), 59, 49, 7591, 7594, 15 Jun. 2023, [International Magazine]
English, Scientific journal, Light-driven inward proton pump rhodopsin RmXeR was embedded in pH-sensitive liposomes. Substance release from the proteoliposomes was observed following light illumination both in vitro and in cells, indicating the successful production of light-induced disruptive liposomes (LiDL). Thus, LiDL is a photoswitchable carrier utilized for intracellular substance delivery.

Differences in the Intracellular Localization of Methylated β-Cyclodextrins-Threaded Polyrotaxanes Lead to Different Cellular States
Yuma Yamada, Shinnosuke Daikuhara, Atsushi Tamura, Kei Nishida, Nobuhiko Yui, Hideyoshi Harashima
Biomolecules, 13, 6, 903, 903, MDPI AG, 29 May 2023
Scientific journal, Activation of autophagy represents a potential therapeutic strategy for the treatment of diseases that are caused by the accumulation of defective proteins and the formation of abnormal organelles. Methylated β-cyclodextrins-threaded polyrotaxane (Me-PRX), a supramolecular structured polymer, induces autophagy by interacting with the endoplasmic reticulum. We previously reported on the successful activation of mitochondria-targeted autophagy by delivering Me-RRX to mitochondria using a MITO-Porter, a mitochondria-targeted nanocarrier. The same level of autophagy induction was achieved at one-twentieth the dosage for the MITO-Porter (Me-PRX) compared to the naked Me-PRX. We report herein on the quantitative evaluation of the intracellular organelle localization of both naked Me-PRX and the MITO-Porter (Me-PRX). Mitochondria, endoplasmic reticulum and lysosomes were selected as target organelles because they would be involved in autophagy induction. In addition, organelle injury and cell viability assays were performed. The results showed that the naked Me-PRX and the MITO-Porter (Me-PRX) were localized in different intracellular organelles, and organelle injury was different, depending on the route of administration, indicating that different organelles contribute to autophagy induction. These findings indicate that the organelle to which the autophagy-inducing molecules are delivered plays an important role in the level of induction of autophagy.

A system that delivers an antioxidant to mitochondria for the treatment of drug-induced liver injury.
Mitsue Hibino, Masatoshi Maeki, Manabu Tokeshi, Yoichi Ishitsuka, Hideyoshi Harashima, Yuma Yamada
Scientific reports, 13, 1, 6961, 6961, 10 May 2023, [Peer-reviewed], [International Magazine]
English, Scientific journal, Mitochondria, a major source of reactive oxygen species (ROS), are intimately involved in the response to oxidative stress in the body. The production of excessive ROS affects the balance between oxidative responses and antioxidant defense mechanisms thus perturbing mitochondrial function eventually leading to tissue injury. Therefore, antioxidant therapies that target mitochondria can be used to treat such diseases and improve general health. This study reports on an attempt to establish a system for delivering an antioxidant molecule coenzyme Q10 (CoQ10) to mitochondria and the validation of its therapeutic efficacy in a model of acetaminophen (APAP) liver injury caused by oxidative stress in mitochondria. A CoQ10-MITO-Porter, a mitochondrial targeting lipid nanoparticle (LNP) containing encapsulated CoQ10, was prepared using a microfluidic device. It was essential to include polyethylene glycol (PEG) in the lipid composition of this LNP to ensure stability of the CoQ10, since it is relatively insoluble in water. Based on transmission electron microscope (TEM) observations and small angle X-ray scattering (SAXS) measurements, the CoQ10-MITO-Porter was estimated to be a 50 nm spherical particle without a regular layer structure. The use of the CoQ10-MITO-Porter improved liver function and reduced tissue injury, suggesting that it exerted a therapeutic effect on APAP liver injury.

Delivering mRNA to a human NK cell line, NK-92 cells, by lipid nanoparticles.
Takashi Nakamura, Taisei Nakade, Yusuke Sato, Hideyoshi Harashima
International journal of pharmaceutics, 636, 122810, 122810, 05 Apr. 2023, [International Magazine]
English, Scientific journal, In cancer immunotherapy, therapeutic methods targeting NK are highly expected. NK cell-based therapy using NK-92, a human NK cell line, has been clinically evaluated. Delivering mRNA into NK-92 cells is a potent strategy for enhancing its functions. However, the use of lipid nanoparticles (LNP) for this purpose has not yet been evaluated. We previously developed a LNP that was composed of CL1H6 (CL1H6-LNP) for the efficient delivery of siRNA to NK-92 cells, and the use of this material for delivering mRNA to NK-92 cells is reported in this study. Compared with a DLin-MC3-DMA based LNP, used as a benchmark, the CL1H6-LNP caused a high mRNA expression intensity and a cell transfection efficiency of 100%. The efficient mRNA delivery by this CL1H6-LNP is attributed to the high affinity for NK-92 cells and the intense, rapid fusion with the endosomal membrane. It therefore appears that the CL1H6-LNP could be a useful non-viral vector for modifying the NK-92 functions by mRNA. Our findings also provide some insights into the design and development of LNPs for delivering mRNA to NK-92 and NK cells.

Microenvironmental Changes in Mediastinal Fat-associated Lymphoid Clusters and Lungs in Early and Late Stages of Metastatic Lung Cancer Induction
Yaser Hosny Ali Elewa, Mahmoud M Abd Elwakil, Hideyoshi Harashima, Sherif Kh A Mohamed, Mahmoud Hosny Zahran
Microscopy and Microanalysis, 29, 3, 1228, 1243, Oxford University Press (OUP), 01 Apr. 2023
Scientific journal, Abstract

The prognosis of metastatic lung melanoma (MLM) has been reported to be poor. An increasing number of studies have reported the function of several immune cells in cancer regression. Although the function of mediastinal fat-associated lymphoid clusters (MFALCs) in the progression of inflammatory lung lesions has been previously reported, the association between MLM progression and MFALCs development has remained unexplored. Herein, we compared the microenvironmental changes in the lungs and MFALCs among phosphate-buffered saline (PBS) and cancer groups at early (1 week) and late (2 weeks) stages following the intravenous injection of B16-F10 melanoma cells into C57BL/6 mice. Except for lung CD4+ helper T-cells and Iba1+ macrophage populations of early stage, we observed a significant increase in the proliferating and immune cell (CD20+ B-lymphocytes, CD3+ T-lymphocytes, CD8+ cytotoxic T-cells, CD16+ natural killer (NK) cells populations, area of high endothelial venules, and lung lymphatic vessels in cancer groups at both the stages as compared with the PBS groups. Furthermore, a significant positive correlation was observed between immune cell populations in MFALCs and the lungs (B- and T-lymphocytes, and NK cells in both stages). Collectively, our findings suggest a promising cancer therapeutic strategy via targeting immune cells in MFALCs.

Mass production system for RNA-loaded lipid nanoparticles using piling up microfluidic devices
Masatoshi Maeki, Yuto Okada, Shuya Uno, Kaisei Sugiura, Yuichi Suzuki, Kento Okuda, Yusuke Sato, Masao Ando, Hiroyuki Yamazaki, Masaki Takeuchi, Akihiko Ishida, Hirofumi Tani, Hideyoshi Harashima, Manabu Tokeshi
Applied Materials Today, Apr. 2023, [Peer-reviewed]
Scientific journal

Development of a Mitochondrial Targeting Lipid Nanoparticle Encapsulating Berberine
Ikuma Hori, Hideyoshi Harashima, Yuma Yamada
International Journal of Molecular Sciences, 24, 2, 903, 903, MDPI AG, 04 Jan. 2023
Scientific journal, Delivering drugs to mitochondria, the main source of energy in neurons, can be a useful therapeutic strategy for the treatment of neurodegenerative diseases. Berberine (BBR), an isoquinoline alkaloid, acts on mitochondria and is involved in mechanisms associated with the normalization and regulation of intracellular metabolism. Therefore, BBR has attracted considerable interest as a possible therapeutic drug for neurodegenerative diseases. While BBR has been reported to act on mitochondria, there are few reports on the efficient delivery of BBR into mitochondria. This paper reports on the mitochondrial delivery of BBR using a lipid nanoparticle (LNP), a “MITO-Porter” that targets mitochondria, and its pharmacological action in Neuro2a cells, a model neuroblastoma. A MITO-Porter containing encapsulated BBR (MITO-Porter (BBR)) was prepared. Treatment with MITO-Porter (BBR) increased the amount of BBR that accumulated in mitochondria compared with a treatment with naked BBR. Treatment with MITO-Porter (BBR) resulted in increased ATP production in Neuro2a cells, which are important for maintaining life phenomena, compared with treatment with naked BBR. Treatment with MITO-Porter (BBR) also increased the level of expression of mitochondrial ubiquitin ligase (MITOL), which is involved in mitochondrial quality control. Our findings indicate that increasing the accumulation of BBR into mitochondria is important for inducing enhanced pharmacological actions. The use of this system has the potential for being important in terms of the regulation of the metabolic mechanism of mitochondria in nerve cells.

Self-homing nanocarriers for mRNA delivery to the activated hepatic stellate cells in liver fibrosis
Mahmoud A. Younis, Yusuke Sato, Yaser H.A. Elewa, Yasuhiro Kon, Hideyoshi Harashima
Journal of Controlled Release, 353, 685, 698, Elsevier BV, Jan. 2023
Scientific journal

Fine-tuning the encapsulation of a photosensitizer in nanoparticles reveals the relationship between internal structure and phototherapeutic effects.
Fumika Kubota, Satrialdi, Yuta Takano, Masatoshi Maeki, Manabu Tokeshi, Hideyoshi Harashima, Yuma Yamada
Journal of biophotonics, 16, 3, e202200119, 01 Sep. 2022, [International Magazine]
English, Scientific journal, Photodynamic therapy (PDT) is a cancer therapy that uses a photosensitizer (PS) in the presence of oxygen molecules. Since singlet oxygen is highly reactive, it is important to deliver it to the target site. Thus, an efficient drug delivery system (DDS) is essential for enhancing the efficacy of such a treatment and protecting against the side effects of PDT. Here, we report on attempts to increase the therapeutic effect of PDT by using a DDS, a lipid nanoparticle (LNP). We prepared a porphyrin analog, rTPA (PS) that was encapsulated in LNPs using a microfluidic device. The findings indicated that the internal structure of the prepared particles changed depending on the amount of rTPA in LNPs. The photoactivity and cell-killing effect of PS in LNPs also changed when the amount of the cargo increased. These results suggest that the internal structure of LNPs is important factors that affect drug efficacy. This article is protected by copyright. All rights reserved.

Interval- and cycle-dependent combined effect of STING agonist loaded lipid nanoparticles and a PD-1 antibody.
Alaa M Khalifa, Takashi Nakamura, Yusuke Sato, Takanori Sato, Mamoru Hyodo, Yoshihiro Hayakawa, Hideyoshi Harashima
International journal of pharmaceutics, 624, 122034, 122034, 25 Aug. 2022, [International Magazine]
English, Scientific journal, Programmed cell death 1 (PD-1) blockade combination to other drugs have attracted the interest of scientists for treating tumors resistant to PD-1 blockade. In this study, the impact of the interval, order of administration, and number of cycles of immunotherapeutic combination of stimulator of interferon genes (STING) pathway agonist loaded lipid nanoparticle (STING-LNP) and PD-1 antibody for inducing the optimal combined antitumor activity against a melanoma lung metastasis is reported. One cycle had no effect, but two and three cycles resulted in a combinedantitumor effect. The interval between the administration was found to influence the induction of the combined effect. The second and third doses increased the gene expression of the NK cell activation marker, interferon γ (IFN-γ), PD-1 and a ligand of PD-1 (PD-L1), whereas the first dose failed. NK cells in the lung showed an increase in the expression of the activation markers and PD-1 after the second dose. The combined antitumor effect of this combination therapy against melanoma lung metastasis model could be dependent on the interval as well as the number of doses of STING-LNP.These findings suggest the importance of the protocol setting when combining a nano system loaded with an immune adjuvant and PD-1 antibody.

Inhibitory role of Annexin A1 in pathological bone resorption and therapeutic implications in periprosthetic osteolysis.
Hend Alhasan, Mohamad Alaa Terkawi, Gen Matsumae, Taku Ebata, Yuan Tian, Tomohiro Shimizu, Yoshio Nishida, Shunichi Yokota, Fayna Garcia-Martin, Mahmoud M Abd Elwakil, Daisuke Takahashi, Mahmoud A Younis, Hideyoshi Harashima, Ken Kadoya, Norimasa Iwasaki
Nature communications, 13, 1, 3919, 3919, 07 Jul. 2022, [International Magazine]
English, Scientific journal, There is currently no therapy available for periprosthetic osteolysis, the most common cause of arthroplasty failure. Here, the role of AnxA1 in periprosthetic osteolysis and potential therapeutics were investigated. Reducing the expression of AnxA1 in calvarial tissue was found to be associated with increased osteolytic lesions and the osteolytic lesions induced by debris implantation were more severe in AnxA1-defecient mice than in wild-type mice. AnxA1 inhibits the differentiation of osteoclasts through suppressing NFκB signaling and promoting the PPAR-γ pathway. Administration of N-terminal-AnxA1 (Ac2-26 peptide) onto calvariae significantly reduced osteolytic lesions triggered by wear debris. These therapeutic effects were abrogated in mice that had received the PPAR-γ antagonist, suggesting that the AnxA1/PPAR-γ axis has an inhibitory role in osteolysis. The administration of Ac2-26 suppressed osteolysis induced by TNF-α and RANKL injections in mice. These findings indicate that AnxA1 is a potential therapeutic agent for the treatment of periprosthetic osteolysis.

Extrahepatic targeting of lipid nanoparticles in vivo with intracellular targeting for future nanomedicines.
Takashi Nakamura, Yusuke Sato, Yuma Yamada, Mahmoud M Abd Elwakil, Seigo Kimura, Mahmoud A Younis, Hideyoshi Harashima
Advanced drug delivery reviews, 188, 114417, 114417, 03 Jul. 2022, [International Magazine]
English, Scientific journal, A new era of nanomedicines that involve nucleic acids/gene therapy has been opened after two decades in 21st century and new types of more efficient drug delivery systems (DDS) are highly expected and will include extrahepatic delivery. In this review, we summarize the possibility and expectations for the extrahepatic delivery of small interfering RNA/messenger RNA/plasmid DNA/genome editing to the spleen, lung, tumor, lymph nodes as well as the liver based on our studies as well as reported information. Passive targeting and active targeting are discussed in in vivo delivery and the importance of controlled intracellular trafficking for successful therapeutic results are also discussed. In addition, mitochondrial delivery as a novel strategy for nucleic acids/gene therapy is introduced to expand the therapeutic dimension of nucleic acids/gene therapy in the liver as well as the heart, kidney and brain.

ミトコンドリア活性化心筋幹細胞移植療法の臨床応用に向けた取り組み　　　　　　　　　　　　　　　
白石 真大, 山田 勇磨, 佐々木 大輔, 武田 充人, 原島 秀吉
日本小児循環器学会総会・学術集会抄録集, 58回, [III, 03], (NPO)日本小児循環器学会, Jul. 2022
Japanese

On the size-regulation of RNA-loaded lipid nanoparticles synthesized by microfluidic device.
Kento Okuda, Yusuke Sato, Kazuki Iwakawa, Kosuke Sasaki, Nana Okabe, Masatoshi Maeki, Manabu Tokeshi, Hideyoshi Harashima
Journal of controlled release : official journal of the Controlled Release Society, 348, 648, 659, 18 Jun. 2022, [International Magazine]
English, Scientific journal, The use of lipid nanoparticles (LNPs) for nucleic acid delivery is now becoming a promising strategy with a number of clinical trials as vaccines or as novel therapies against a variety of genetic and infectious diseases. The use of microfluidics for the synthesis of the LNPs has attracted interest because of its considerable advantages over other conventional synthetic methods including scalability, reproducibility, and speed. However, despite the potential usefulness of large particles for nucleic acid delivery to dendritic cells (DCs) as a vaccine, the particle size of the LNPs prepared using microfluidics is typically limited to approximately from 30 to 100 nm. In this study, focusing on Derjaguin-Landau-Verwey-Overbeek (DLVO) theory, the effect of some synthetic parameters, including total flow rate, flow rate ratio, buffer pH, lipid concentration, molar ratio of PEG-lipid as well as salt concentration, on particle size was systematically examined by means of the design of experiment approaches. The findings indicated that the simple addition of salt (e.g. NaCl) to a buffer containing nucleic acids contributed greatly to the synthesis of large LNPs over 200 nm and this effect was concentration-dependent with respect to the salt. The effect of salt on particle size was consistent with a Hofmeister series. The systemic injection of larger mRNA-loaded LNPs resulted in a higher transgene expression in mouse splenic DCs, a higher activation of various splenic immune cells, and had a superior effect as a therapeutic cancer vaccine in a syngeneic mouse model compared to the smaller-sized counterpart with constant lipid composition prepared with lower NaCl concentration. Collectively, size-regulation by the simple addition of salt is a promising strategy for developing potent LNPs.

Retrograde Axonal Transport of Liposomes from Peripheral Tissue to Spinal Cord and DRGs by Optimized Phospholipid and CTB Modification.
Takafumi Fukui, Hironao Tateno, Takashi Nakamura, Yuma Yamada, Yusuke Sato, Norimasa Iwasaki, Hideyoshi Harashima, Ken Kadoya
International journal of molecular sciences, 23, 12, 15 Jun. 2022, [International Magazine]
English, Scientific journal, Despite recent advancements in therapeutic options for disorders of the central nervous system (CNS), the lack of an efficient drug-delivery system (DDS) hampers their clinical application. We hypothesized that liposomes could be optimized for retrograde transport in axons as a DDS from peripheral tissues to the spinal cord and dorsal root ganglia (DRGs). Three types of liposomes consisting of DSPC, DSPC/POPC, or POPC in combination with cholesterol (Chol) and polyethylene glycol (PEG) lipid were administered to sciatic nerves or the tibialis anterior muscle of mature rats. Liposomes in cell bodies were detected with infrared fluorescence of DiD conjugated to liposomes. Three days later, all nerve-administered liposomes were retrogradely transported to the spinal cord and DRGs, whereas only muscle-administered liposomes consisting of DSPC reached the spinal cord and DRGs. Modification with Cholera toxin B subunit improved the transport efficiency of liposomes to the spinal cord and DRGs from 4.5% to 17.3% and from 3.9% to 14.3% via nerve administration, and from 2.6% to 4.8% and from 2.3% to 4.1% via muscle administration, respectively. Modification with octa-arginine (R8) improved the transport efficiency via nerve administration but abolished the transport capability via muscle administration. These findings provide the initial data for the development of a novel DDS targeting the spinal cord and DRGs via peripheral administration.

Innovative cancer nanomedicine based on immunology, gene editing, intracellular trafficking control.
Yuma Yamada, Yusuke Sato, Takashi Nakamura, Hideyoshi Harashima
Journal of controlled release : official journal of the Controlled Release Society, 348, 357, 369, 09 Jun. 2022, [International Magazine]
English, Scientific journal, The recent rapid progress in the area of drug delivery systems (DDS) has opened a new era in medicine with a strong linkage to understanding the molecular mechanisms associated with cancer survival. In this review, we summarize new cancer strategies that have recently been developed based on our DDS technology. Cancer immunotherapy will be improved based on the concept of the cancer immunity cycle, which focuses on dynamic interactions between various types of cancer and immune cells in our body. The new technology of genome editing will also be discussed with reference to how these new DDS technologies can be used to introduce therapeutic cargoes into our body. Lastly, a new organelle, mitochondria will be the focus of creating a new cancer treatment strategy by a MITO-Porter which can deliver macromolecules directly to mitochondria of cancer cells via a membrane fusion approach and the impact of controlled intracellular trafficking will be discussed.

Validation of a therapeutic strategy involving the mitochondrial delivery of thiamine pyrophosphate using brain damage induced mouse model
Yuma Yamada, Takuya Ishimaru, Hideyoshi Harashima
Clinical and Translational Discovery, 2, 2, Wiley, Jun. 2022
Scientific journal

Recent advances in delivering RNA-based therapeutics to mitochondria.
Yuma Yamada, Sen Ishizuka, Manae Arai, Minako Maruyama, Hideyoshi Harashima
Expert opinion on biological therapy, 1, 11, 11 May 2022, [International Magazine]
English, Scientific journal, INTRODUCTION: After the emergence of lipid nanoparticles (LNP) containing therapeutic mRNA as vaccines for use against the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the clinical usefulness of nucleic acid-encapsulated LNPs is now a fact. In addition to the nucleus and cytoplasm, mitochondria, which have their own genome, are a site where nucleic acids function in the cell. Gene therapies targeting mitochondria are expected to pave the way for the next generation of therapies. AREAS COVERED: Methods for delivering nucleic acids to mitochondria are needed in order to realize such innovative therapies. However, only a few reports on delivery systems targeting mitochondria have appeared. In this review, we summarize the current state of research on RNA-based therapeutics targeted to mitochondria, with emphasis on mitochondrial RNA delivery therapies and on therapies that involve the use of mitochondrial genome editing devices. EXPERT OPINION: We hope that this review article will focus our attention to this area of research, stimulate more interest in this field of research, and lead to the development of mitochondria-targeted nucleic acid medicine. It has the potential to become a major weapon against urgent and unknown diseases, including SARS-CoV-2 infections.

Combined nano cancer immunotherapy based on immune status in a tumor microenvironment.
Takashi Nakamura, Kyoko Kawakami, Momoka Nomura, Yusuke Sato, Mamoru Hyodo, Hiroto Hatakeyama, Yoshihiro Hayakawa, Hideyoshi Harashima
Journal of controlled release : official journal of the Controlled Release Society, 345, 200, 213, May 2022, [International Magazine]
English, Scientific journal, Since the effect of cancer immunotherapy is largely dependent on the status of the immune system in the tumor microenvironment (TME), choice of therapy and the development of new therapies based on the immune status in the TME would be predicted to be effective. Unfortunately, the development of delivery systems for such therapy has been slow. Here, we defined a parameter of immune status in TME showing antitumor effects and demonstrated the cancer immunotherapy with an adjuvant loaded lipid nanoparticle (LNP), which was taken advantage the parameter. An analysis was carried out to determine the relationship between antitumor effects and gene expression (22 target genes) in tumors (MC38 and E.G7-OVA) that respond to the programmed cell death 1 (PD-1) antibody and non-responding tumors (B16-F10 and 4T1). The immune status showing an effective antitumor effect, which consisted of 10 genes, was then extracted. Treatment with the adjuvant loaded LNP caused a significant antitumor effect against an E.G7-OVA tumor, and the gene expression in the E.G7-OVA tumor was completely within the range of gene expression for showing an effective antitumor effect, as defined by the identified immune status panel (IS-panel-10). Although the treatment with the adjuvant loaded LNP failed to induce a sufficient antitumor effect against the 4T1 tumor, we succeeded in enhancing the antitumor effect by using a combination therapy that was adopted based on the analysis by the IS-panel-10 in the TME. The 10 genes were found to affect the prognosis in a variety of human cancers. Collectively, the findings reported herein demonstrate the potential of immune status analysis in the TME for developing cancer immunotherapies using a delivery system.

Non-invasive gene delivery across the blood-brain barrier: present and future perspectives.
Seigo Kimura, Hideyoshi Harashima
Neural regeneration research, 17, 4, 785, 787, Apr. 2022, [Peer-reviewed], [Invited], [Last author], [International Magazine]
English, Scientific journal

Transplantation of MITO cells, mitochondria activated cardiac progenitor cells, to the ischemic myocardium of mouse enhances the therapeutic effect.
Daisuke Sasaki, Jiro Abe, Atsuhito Takeda, Hideyoshi Harashima, Yuma Yamada
Scientific reports, 12, 1, 4344, 4344, 22 Mar. 2022, [International Magazine]
English, Scientific journal, Given the potential for myocardial stem cell transplantation as a promising treatment for heart failure, numerous clinical trials have been conducted and its usefulness has been clearly confirmed. However, the low rate of engraftment of transplanted cells has become a clinical problem, and this needs to be improved in the case of transplanting cells to the heart. To address this issue, we report on attempts to prepare mitochondria-activated stem cells (MITO cells) for use in transplantation. MITO cells, which is cardiac progenitor cells (CPCs) activated by the mitochondrial delivery of resveratrol with an anti-oxidant and mitochondrial activation effects were successfully prepared using a mitochondrial targeting nanocarrier (MITO-Porter). The purpose of this study was to validate the therapeutic effect of cell transplantation by the MITO cells using a mouse model of myocardial ischemia-reperfusion. Mouse CPCs were used as transplanted cells. The transplantation of CPCs and MITO cells were conducted after myocardial ischemia-reperfusion, and the therapeutic effect was determined. The MITO cells transplanted group showed increase in postoperative weight gain, improve cardiac function and inhibition of fibrosis compared to the non-transplanted group and the CPC group. The transplantation of MITO cells to the ischemic myocardium showed a stronger transplantation effect compared to conventional CPC transplantation.

Novel antiangiogenic therapy targeting biglycan using tumor endothelial cell-specific liposomal siRNA delivery system.
Nako Maishi, Yu Sakurai, Hiroto Hatakeyama, Yui Umeyama, Takashi Nakamura, Rikito Endo, Mohammad Towfik Alam, Cong Li, Dorcas Akuba-Muhyia Annan, Hiroshi Kikuchi, Hirofumi Morimoto, Masahiro Morimoto, Kosuke Akiyama, Noritaka Ohga, Yasuhiro Hida, Hideyoshi Harashima, Kyoko Hida
Cancer science, 113, 5, 1855, 1867, 09 Mar. 2022, [International Magazine]
English, Scientific journal, Tumor blood vessels play important roles in tumor progression and metastasis. Targeting tumor endothelial cells (TECs) is one of the strategies for cancer therapy. We previously reported that biglycan, a small leucine-rich proteoglycan, is highly expressed in TECs. TECs utilize biglycan in an autocrine manner for migration and angiogenesis. Furthermore, TEC-derived biglycan stimulates tumor cell migration in a paracrine manner leading to tumor cell intravasation and metastasis. In this study, we explored the therapeutic effect of biglycan inhibition in the TECs of renal cell carcinoma using an in vivo siRNA delivery system known as a multifunctional envelope-type nanodevice (MEND), which contains a unique pH-sensitive cationic lipid. To specifically deliver MEND into TECs, we incorporated cyclo(Arg-Gly-Asp-D-Phe-Lys) (cRGD) into MEND because αV β3 integrin, a receptor for cRGD, is selective and highly expressed in TECs. We developed RGD-MEND-encapsulating siRNA against biglycan. First, we confirmed that MEND was delivered into OS-RC-2 tumor-derived TECs and induced in vitro RNAi-mediated gene silencing. MEND was then injected intravenously into OS-RC-2 tumor-bearing mice. Flow cytometry analysis demonstrated that MEND was specifically delivered into TECs. Quantitative RT-PCR indicated that biglycan was knocked down by biglycan siRNA-containing MEND. Finally, we analyzed the therapeutic effect of biglycan silencing by MEND in TECs. Tumor growth was inhibited by biglycan siRNA-containing MEND. Tumor microenvironmental factors such as fibrosis were also normalized using biglycan inhibition in TECs. Biglycan in TECs can be a novel target for cancer treatment.

ミトコンドリア標的型DDSによるミトコンドリア病治療戦略の検証　　　　　　　　　　　　　　　
荒井 愛永, 山田 勇磨, 佐々木 大輔, 真栄城 正寿, 渡慶次 学, 原島 秀吉
日本薬学会年会要旨集, 142年会, 27I, am02S, (公社)日本薬学会, Mar. 2022
Japanese

Strategies for fighting pandemic virus infections: Integration of virology and drug delivery.
Takashi Nakamura, Norikazu Isoda, Yoshihiro Sakoda, Hideyoshi Harashima
Journal of controlled release : official journal of the Controlled Release Society, 343, 361, 378, 03 Feb. 2022, [Peer-reviewed], [International Magazine]
English, Scientific journal, Respiratory viruses have sometimes resulted in worldwide pandemics, with the influenza virus and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) being major participants. Long-term efforts have made it possible to control the influenza virus, but seasonal influenza continues to take many lives each year, and a pandemic influenza virus sometimes emerges. Although vaccines for coronavirus disease 2019 (COVID-19) have been developed, we are not yet able to coexist with the SARS-CoV-2. To overcome such viruses, it is necessary to obtain knowledge about international surveillance systems, virology, ecology and to determine that immune responses are effective. The information must then be transferred to drugs. Delivery systems would be expected to contribute to the rational development of drugs. In this review, virologist and drug delivery system (DDS) researchers discuss drug delivery strategies, especially the use of lipid-based nanocarriers, for fighting to respiratory virus infections.

Resveratrol-Encapsulated Mitochondria-Targeting Liposome Enhances Mitochondrial Respiratory Capacity in Myocardial Cells.
Takao Tsujioka, Daisuke Sasaki, Atsuhito Takeda, Hideyoshi Harashima, Yuma Yamada
International journal of molecular sciences, 23, 1, 22 Dec. 2021, [International Magazine]
English, Scientific journal, The development of drug delivery systems for use in the treatment of cardiovascular diseases is an area of great interest. We report herein on an evaluation of the therapeutic potential of a myocardial mitochondria-targeting liposome, a multifunctional envelope-type nano device for targeting pancreatic β cells (β-MEND) that was previously developed in our laboratory. Resveratrol (RES), a natural polyphenol compound that has a cardioprotective effect, was encapsulated in the β-MEND (β-MEND (RES)), and its efficacy was evaluated using rat myocardioblasts (H9c2 cells). The β-MEND (RES) was readily taken up by H9c2 cells, as verified by fluorescence-activated cell sorter data, and was observed to be colocalized with intracellular mitochondria by confocal laser scanning microscopy. Myocardial mitochondrial function was evaluated by a Seahorse XF Analyzer and the results showed that the β-MEND (RES) significantly activated cellular maximal respiratory capacity. In addition, the β-MEND (RES) showed no cellular toxicity for H9c2 cells as evidenced by Premix WST-1 assays. This is the first report of the use of a myocardial mitochondria-targeting liposome encapsulating RES for activating mitochondrial function, which was clearly confirmed based on analyses using a Seahorse XF Analyzer.

An effective in vivo mitochondria-targeting nanocarrier combined with a pi-extended porphyrin-type photosensitizer
Satrialdi, Yuta Takano, Eri Hirata, Natsumi Ushijima, Hideyoshi Harashima, Yuma Yamada
NANOSCALE ADVANCES, 3, 20, 5919, 5927, Oct. 2021
English, Scientific journal

The hydrophobic tail of a pH-sensitive cationic lipid influences siRNA transfection activity and toxicity in human NK cell lines.
Takashi Nakamura, Taisei Nakade, Koharu Yamada, Yusuke Sato, Hideyoshi Harashima
International journal of pharmaceutics, 609, 121140, 121140, 27 Sep. 2021, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, The use of natural killer (NK) cells in cell therapy is an attractive next generation strategy for cancer immunotherapy. NK-92 cells (a human NK cell line) have been tested in clinical trial stages, making them an off-the-shelf medicine. Controlling gene expression in NK-92 cells by an artificial delivery system is an available for enhancing NK-92 cell therapy. We report here on the development of a siRNA-loaded lipid nanoparticle (LNP) composed of CL1H6 (CL1H6-LNP), an optimized, pH-sensitive cationic lipid, with efficient gene silencing and low cytotoxicity in NK-92 cells. The hydrophilic head group of the lipid molecule used in preparing these particles largely influences the pKa of the final LNP, and lipids with an amino moiety substituted with a methyl group showed a high gene silencing activity. Compared with myristate and palmitate, the hydrophobic tail of oleate had a high gene silencing activity and cell viability. Analyses of intracellular trafficking indicated that the CL1H6-LNP appeared to escape from the endosomes via membrane fusion, without disrupting the membrane. The mechanism of endosomal escape should contribute to our understanding of efficient gene silencing with a low degree of cytotoxicity. These results therefore suggest that a CL1H6-LNP has promise for delivering siRNA to NK-92 cells.

Maximizing the Oral Bioavailability of Poorly Water-Soluble Drugs Using Novel Oil-Like Materials in Lipid-Based Formulations.
Saed Abbasi, Haruki Higashino, Yusuke Sato, Keiko Minami, Makoto Kataoka, Shinji Yamashita, Hideyoshi Harashima
Molecular pharmaceutics, 18, 9, 3281, 3289, 06 Sep. 2021, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Lipid-based formulations, such as self-microemulsifying drug-delivery systems (SMEDDSs), are promising tools for the oral delivery of poorly water-soluble drugs. However, failure to maintain adequate aqueous solubility after coming into contact with gastrointestinal fluids is a major drawback. In this study, we examined the use of a novel cinnamic acid-derived oil-like material (CAOM) that binds drugs with a high affinity through π-π stacking and hydrophobic interactions, as an oil core in a SMEDDS for the oral delivery of fenofibrate in rats. The use of the CAOM in the SMEDDS resulted in an unprecedented enhancement in fenofibrate bioavailability, which exceeded the bioavailability values obtained using SMEDDSs based on corn oil, a conventional triglyceride oil, or Labrasol, an enhancer of intestinal permeation. Further characterization revealed that the CAOM SMEDDS does not alter the intestinal permeability and has no inhibitory activity on P-glycoprotein-mediated drug efflux. The results reported herein demonstrate the strong potential of CAOM formulations as new solubilizers for the efficient and safe oral delivery of drugs that have limited water solubility.

Validation of the mitochondrial delivery of vitamin B1 to enhance ATP production using SH-SY5Y cells, a model neuroblast.
Yuma Yamada, Takuya Ishimaru, Kohei Ikeda, Hideyoshi Harashima
Journal of pharmaceutical sciences, 111, 2, 432, 439, 31 Aug. 2021, [Peer-reviewed], [Last author], [International Magazine]
English, Scientific journal, Large amounts of ATP are produced in mitochondria especially in the brain and heart, where energy consumption is high compared with other organs. Thus, a decrease in ATP production in such organs could be a cause of many diseases such as neurodegenerative diseases and heart disease. Based on thus assumption, increasing intracellular ATP production in such organs could be a therapeutic strategy. In this study, we report on the delivery of vitamin B1, a coenzyme that activates the tricarboxylic acid (TCA) cycle, to the inside of mitochondria. Since the TCA cycle is responsible for ATP production, we hypothesized delivering vitamin B1 to mitochondria would enhance ATP production. To accomplish this, we used a mitochondrial targeted liposome a "MITO-Porter" as the carrier. Using SH-SY5Y cells, a model neuroblast, cellular uptake and intracellular localization were analyzed using flow cytometry and confocal laser scanning microscopy. The optimized MITO-Porter containing encapsulated vitamin B1 (MITO-Porter (VB1)) was efficiently accumulated in mitochondria of SH-SY5Y cells. Further studies confirmed that the level of ATP production after the MITO-Porter (VB1) treatment was significantly increased as compared to a control group that was treated with naked vitamin B1. This study provides the potential for an innovative therapeutic strategy in which the TCA cycle is activated, thus enhancing ATP production. Relative ATP ratio (%) = IS/IU × 100, where IS and IU represent the intracellular ATP amounts for the treated and untreated cells with samples, respectively.

イメージングと操作で見るミトコンドリア機能 ミトコンドリア機能を操作するDrug Delivery System　　　　　　　　　　　　　　　
山田 勇磨, 原島 秀吉
バイオイメージング, 30, 2, 38, 38, 日本バイオイメージング学会, Aug. 2021
Japanese

心筋症の基礎と臨床の架け橋を探る ヒト心筋前駆細胞を用いたミトコンドリア活性化心筋前駆細胞の製造および、心筋虚血再灌流モデルに対する細胞移植の治療効果の検討　　　　　　　　　　　　　　　
佐々木 大輔, 山田 勇磨, 後藤 悠太, 白石 真大, 武田 充人, 原島 秀吉
日本小児循環器学会総会・学術集会抄録集, 57回, [I, 5], (NPO)日本小児循環器学会, Jul. 2021
Japanese

心筋ミトコンドリアを標的としたリポソーム製剤の開発研究　　　　　　　　　　　　　　　
辻岡 孝郎, 山田 勇磨, 佐々木 大輔, 武田 充人, 原島 秀吉
日本小児循環器学会総会・学術集会抄録集, 57回, [OR31, 1], (NPO)日本小児循環器学会, Jul. 2021
Japanese

STING agonist loaded lipid nanoparticles overcome anti-PD-1 resistance in melanoma lung metastasis via NK cell activation.
Takashi Nakamura, Takanori Sato, Rikito Endo, Shun Sasaki, Naomichi Takahashi, Yusuke Sato, Mamoru Hyodo, Yoshihiro Hayakawa, Hideyoshi Harashima
Journal for immunotherapy of cancer, 9, 7, Jul. 2021, [Peer-reviewed], [Last author], [International Magazine]
English, Scientific journal, BACKGROUND: Resistance to an immune checkpoint inhibitor (ICI) is a major obstacle in cancer immunotherapy. The causes of ICI resistance include major histocompatibility complex (MHC)/histocompatibility locus antigen (HLA) class I loss, neoantigen loss, and incomplete antigen presentation. Elimination by natural killer (NK) cells would be expected to be an effective strategy for the treatment of these ICI-resistant tumors. We previously demonstrated that a lipid nanoparticle containing a stimulator of an interferon gene (STING) agonist (STING-LNP) efficiently induced antitumor activity via the activation of NK cells. Thus, we evaluated the potential of reducing ICI resistance by STING-LNPs. METHODS: Lung metastasis of a B16-F10 mouse melanoma was used as an anti-programmed cell death 1 (anti-PD-1)-resistant mouse model. The mice were intravenously injected with the STING-LNP and the mechanism responsible for the improvement of anti-PD-1 resistance by the STING-LNPs was analyzed by RT-qPCR and flow cytometry. The dynamics of STING-LNP were also investigated. RESULTS: Although anti-PD-1 monotherapy failed to induce an antitumor effect, the combination of the STING-LNP and anti-PD-1 exerted a synergistic antitumor effect. Our results indicate that the STING-LNP treatment significantly increased the expression of CD3, CD4, NK1.1, PD-1 and interferon (IFN)-γ in lung metastases. This change appears to be initiated by the type I IFN produced by liver macrophages that contain the internalized STING-LNPs, leading to the systemic activation of NK cells that express PD-1. The activated NK cells appeared to produce IFN-γ, resulting in an increase in the expression of the PD ligand 1 (PD-L1) in cancer cells, thus leading to a synergistic antitumor effect when anti-PD-1 is administered. CONCLUSIONS: We provide a demonstration to show that a STING-LNP treatment can overcome PD-1 resistance in a B16-F10 lung metastasis model. The mechanism responsible for this indicates that NK cells are activated by stimulating the STING pathway which, in turn, induced the expression of PD-L1 on cancer cells. Based on the findings reported herein, the STING-LNP represents a promising candidate for use in combination therapy with anti-PD-1-resistant tumors.

ミトコンドリア人工共生による細胞機能制御の試み　　　　　　　　　　　　　　　
山田 勇磨, 日比野 光恵, 伊藤 百, 荒井 愛永, 佐々木 大輔, 真栄城 正寿, 渡慶次 学, 太田 善浩, 原島 秀吉
日本DDS学会学術集会プログラム予稿集, 37回, 111, 111, 日本DDS学会, Jun. 2021
Japanese

心筋虚血再灌流モデルラットに対するヒトミトコンドリア強化心筋幹細胞移植療法の治療効果の検討　　　　　　　　　　　　　　　
佐々木 大輔, 山田 勇磨, 後藤 悠太, 白石 真大, 武田 充人, 原島 秀吉
日本DDS学会学術集会プログラム予稿集, 37回, 99, 99, 日本DDS学会, Jun. 2021
Japanese

心筋虚血再灌流モデルラットに対するヒトミトコンドリア強化心筋幹細胞移植療法の治療効果の検討　　　　　　　　　　　　　　　
佐々木 大輔, 山田 勇磨, 後藤 悠太, 白石 真大, 武田 充人, 原島 秀吉
日本DDS学会学術集会プログラム予稿集, 37回, 99, 99, 日本DDS学会, Jun. 2021
Japanese

ミトコンドリア人工共生による細胞機能制御の試み　　　　　　　　　　　　　　　
山田 勇磨, 日比野 光恵, 伊藤 百, 荒井 愛永, 佐々木 大輔, 真栄城 正寿, 渡慶次 学, 太田 善浩, 原島 秀吉
日本DDS学会学術集会プログラム予稿集, 37回, 111, 111, 日本DDS学会, Jun. 2021
Japanese

Interferon signaling suppresses the unfolded protein response and induces cell death in hepatocytes accumulating hepatitis B surface antigen.
Ian Baudi, Masanori Isogawa, Federica Moalli, Masaya Onishi, Keigo Kawashima, Yuji Ishida, Chise Tateno, Yusuke Sato, Hideyoshi Harashima, Hiroyasu Ito, Tetsuya Ishikawa, Takaji Wakita, Matteo Iannacone, Yasuhito Tanaka
PLoS pathogens, 17, 5, e1009228, May 2021, [Peer-reviewed], [International Magazine]
English, Scientific journal, Virus infection, such as hepatitis B virus (HBV), occasionally causes endoplasmic reticulum (ER) stress. The unfolded protein response (UPR) is counteractive machinery to ER stress, and the failure of UPR to cope with ER stress results in cell death. Mechanisms that regulate the balance between ER stress and UPR are poorly understood. Type 1 and type 2 interferons have been implicated in hepatic flares during chronic HBV infection. Here, we examined the interplay between ER stress, UPR, and IFNs using transgenic mice that express hepatitis B surface antigen (HBsAg) (HBs-Tg mice) and humanized-liver chimeric mice infected with HBV. IFNα causes severe and moderate liver injury in HBs-Tg mice and HBV infected chimeric mice, respectively. The degree of liver injury is directly correlated with HBsAg levels in the liver, and reduction of HBsAg in the transgenic mice alleviates IFNα mediated liver injury. Analyses of total gene expression and UPR biomarkers' protein expression in the liver revealed that UPR is induced in HBs-Tg mice and HBV infected chimeric mice, indicating that HBsAg accumulation causes ER stress. Notably, IFNα administration transiently suppressed UPR biomarkers before liver injury without affecting intrahepatic HBsAg levels. Furthermore, UPR upregulation by glucose-regulated protein 78 (GRP78) suppression or low dose tunicamycin alleviated IFNα mediated liver injury. These results suggest that IFNα induces ER stress-associated cell death by reducing UPR. IFNγ uses the same mechanism to exert cytotoxicity to HBsAg accumulating hepatocytes. Collectively, our data reveal a previously unknown mechanism of IFN-mediated cell death. This study also identifies UPR as a potential target for regulating ER stress-associated cell death.

Novel PEGylated Lipid Nanoparticles Have a High Encapsulation Efficiency and Effectively Deliver MRTF-B siRNA in Conjunctival Fibroblasts.
Amisha Sanghani, Konstantinos N Kafetzis, Yusuke Sato, Salsabil Elboraie, Julia Fajardo-Sanchez, Hideyoshi Harashima, Aristides D Tagalakis, Cynthia Yu-Wai-Man
Pharmaceutics, 13, 3, 13 Mar. 2021, [Peer-reviewed], [International Magazine]
English, Scientific journal, The master regulator of the fibrosis cascade is the myocardin-related transcription factor/serum response factor (MRTF/SRF) pathway, making it a key target for anti-fibrotic therapeutics. In the past, inhibitors and small interfering RNAs (siRNAs) targeting the MRTF-B gene have been deployed to counter fibrosis in the eye, with the latter showing promising results. However, the biggest challenge in implementing siRNA therapeutics is the method of delivery. In this study, we utilised the novel, pH-sensitive, cationic lipid CL4H6, which has previously demonstrated potent targeting of hepatocytes and endosomal escape, to safely and efficiently deliver an MRTF-B siRNA into human conjunctival fibroblasts. We prepared two lipid nanoparticle (LNP) formulations, incorporating targeting cleavable peptide cY in one of them, and measured their physicochemical properties and silencing effect in human conjunctival fibroblasts. Both proved to be non-cytotoxic at a concentration of 50 nM and effectively silenced the MRTF-B gene in vitro, with the targeting cleavable peptide not affecting the silencing efficiency [LNP with cY: 62.1% and 81.5% versus LNP without cY: 77.7% and 80.2%, at siRNA concentrations of 50 nM (p = 0.06) and 100 nM (p = 0.09), respectively]. On the other hand, the addition of the targeting cleavable peptide significantly increased the encapsulation efficiency of the LNPs from 92.5% to 99.3% (p = 0.0005). In a 3D fibroblast-populated collagen matrix model, both LNP formulations significantly decreased fibroblast contraction after a single transfection. We conclude that the novel PEGylated CL4H6-MRTF-B siRNA-loaded LNPs represent a promising therapeutic approach to prevent conjunctival fibrosis after glaucoma filtration surgery.

Ultra-small lipid nanoparticles encapsulating sorafenib and midkine-siRNA selectively-eradicate sorafenib-resistant hepatocellular carcinoma in vivo.
Mahmoud A Younis, Ikramy A Khalil, Yaser H A Elewa, Yasuhiro Kon, Hideyoshi Harashima
Journal of controlled release : official journal of the Controlled Release Society, 331, 335, 349, 10 Mar. 2021, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Hepatocellular carcinoma (HCC) is a fatal disease with limited therapeutic choices. The stroma-rich tumor microenvironment hinders the in vivo delivery of most nanomedicines. Ultra-small lipid nanoparticles (usLNPs) were designed for the selective co-delivery of the cytotoxic drug, sorafenib (SOR), and siRNA against the Midkine gene (MK-siRNA) to HCC in mice. The usLNPs composed of a novel pH-sensitive lipid, a diversity of phospholipids and a highly-selective targeting peptide. A microfluidic device, iLiNP, was used and a variety of factors were controlled to tune particle size aiming at maximizing tumor penetration efficiency. Optimizing the composition and physico-chemical properties of the usLNPs resulted in an enhanced tumor accumulation, selectivity and in vivo gene silencing. The optimized usLNPs exerted potent gene silencing in the tumor (median effective dose, ED50~0.1 mg/Kg) with limited effect on the healthy liver. The novel combination synergistically-eradicated HCC in mice (~85%) at a surprisingly-low dose of SOR (2.5 mg/Kg) which could not be achieved via individual monotherapy. Toxicity studies revealed the biosafety of the usLNPs upon either acute or chronic treatment. Furthermore, the SOR-resistant HCC established in mice was eradicated by 70% using this approach. We conclude that our strategy is promising for potential clinical applications in HCC treatment.

リポソームを応用した末梢組織から脊髄への低侵襲薬物輸送システムの開発
福井 隆史, 角家 健, 舘野 寛直, 中村 孝司, 山田 勇磨, 佐藤 悠介, 岩崎 倫政, 原島 秀吉
日本整形外科学会雑誌, 95, 2, S123, S123, (公社)日本整形外科学会, Mar. 2021
Japanese

ミトコンドリア強化ヒト心筋幹細胞(hMITO Cell)の製造および心虚血再灌流モデルを用いた細胞移植療法の検証　　　　　　　　　　　　　　　
後藤 悠太, 山田 勇磨, 日比野 光恵, 佐々木 大輔, 武田 充人, 真栄城 正寿, 渡慶次 学, 原島 秀吉
日本薬学会年会要旨集, 141年会, 29V09, pm09S, (公社)日本薬学会, Mar. 2021
Japanese

GALA-Modified Lipid Nanoparticles for the Targeted Delivery of Plasmid DNA to the Lungs.
Yuta Hagino, Ikramy A Khalil, Seigo Kimura, Kenji Kusumoto, Hideyoshi Harashima
Molecular pharmaceutics, 18, 3, 878, 888, 01 Mar. 2021, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, This study describes the development of lipid nanoparticles (LNPs) for the efficient and selective delivery of plasmid DNA (pDNA) to the lungs. The GALA peptide was used as a ligand to target the lung endothelium and as an endosomal escape device. Transfection activity in the lungs was significantly improved when pDNA was encapsulated in double-coated LNPs. The inner coat was composed of dioleoylphsophoethanolamine and a stearylated octaarginine (STR-R8) peptide, while the outer coat was largely a cationic lipid, di-octadecenyl-trimethylammonium propane, mixed with YSK05, a pH-sensitive lipid, and cholesterol. Optimized amounts of YSK05 and GALA were used to achieve an efficient and lung-selective system. The optimized system produced a high gene expression level in the lungs (>107 RLU/mg protein) with high lung/liver and lung/spleen ratios. GALA/R8 modification and the double-coating design were indispensable for efficient gene expression in the lungs. Despite the fact that NPs prepared with 1-step or 2-step coating have the same lipid amount and composition and the same pDNA dose, the transfection activity was dramatically higher in the lungs in the case of 2-step coating. Surprisingly, 1-step or 2-step coatings had no effect on the amount of nanoparticles that were delivered to the lungs, suggesting that the double-coating strategy substantially improved the efficiency of gene expression at the intracellular level.

Lipid nanoparticles loaded with ribonucleoprotein-oligonucleotide complexes synthesized using a microfluidic device exhibit robust genome editing and hepatitis B virus inhibition.
Yuichi Suzuki, Haruno Onuma, Risa Sato, Yusuke Sato, Akari Hashiba, Masatoshi Maeki, Manabu Tokeshi, Mohammad Enamul Hoque Kayesh, Michinori Kohara, Kyoko Tsukiyama-Kohara, Hideyoshi Harashima
Journal of controlled release : official journal of the Controlled Release Society, 330, 61, 71, 10 Feb. 2021, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, The clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) system has considerable therapeutic potential for use in treating a wide range of intractable genetic and infectious diseases including hepatitis B virus (HBV) infections. While non-viral delivery technologies for the CRISPR/Cas system are expected to have clinical applications, difficulties associated with the clinically relevant synthesis of formulations and the poor efficiency of delivery severely hinder therapeutic genome editing. We report herein on the production of a lipid nanoparticle (LNP)-based CRISPR/Cas ribonucleoprotein (RNP) delivery nanoplatform synthesized using a clinically relevant mixer-equipped microfluidic device. DNA cleavage activity and the aggregation of Cas enzymes was completely avoided under the optimized synthetic conditions. The optimized formulation, which was identified through 2 steps of design of experiments, exhibited excellent gene disruption (up to 97%) and base substitution (up to 23%) without any apparent cytotoxicity. The addition of negative charges to the RNPs by complexing single-stranded oligonucleotide (ssON) significantly enhanced the delivery of both Cas9 and Cpf1 RNPs. The optimized formulation significantly suppressed both HBV DNA and covalently closed circular DNA (cccDNA) in HBV-infected human liver cells compared to adeno-associated virus type 2 (AAV2). These findings represent a significant contribution to the development of CRISPR/Cas RNP delivery technology and its practical applications in genome editing therapy.

The nanomedicine rush: New strategies for unmet medical needs based on innovative nano DDS.
Yusuke Sato, Takashi Nakamura, Yuma Yamada, Hideyoshi Harashima
Journal of controlled release : official journal of the Controlled Release Society, 330, 305, 316, 10 Feb. 2021, [Peer-reviewed], [Invited], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, The era of Nanomedicine has arrived with the approval of ONPATTRO™ by the FDA in 2018. Lipid nanoparticle (LNP) technology has succeeded in delivering siRNA to the human liver in genetic diseases and has also been applied to mRNA vaccinations for COVID-19 using a similar LNP technology. In this review, we focus on the current status of new lipids for use in LNP formulations including our original lipids (CL4H6/CL4C6/CL4D6) as well as mechanisms of targeting without a ligand. Clinical applications of nano DDS are moving forward rapidly in the field of cancer immunology since the successful introduction of OPDIVO™ in 2014. Antigen presentation and the maturation of immune cells can be controlled by nano DDS for cancer immunotherapy. YSK12-C4, a newly designed ionizable amino lipid can induce successful immune activation by silencing mRNA in DC and NK cells, which are expected to be evaluated for clinical use. Finally, new cancer therapy by targeting mitochondria involving the use of a MITO-Porter, a membrane fusion-type mitochondrial delivery system, has been introduced. The importance of delivering a photo sensitizer to mitochondria was clearly demonstrated in photodynamic cancer therapy. Clinical applications of MITO-Porters started in collaborative efforts with LUCA Science Co., Ltd. And was established in 2018. The future direction of Nanomedicine is discussed.

Novel lipid combination for delivery of plasmid DNA to immune cells in the spleen.
Seigo Kimura, Ikramy A Khalil, Yaser H A Elewa, Hideyoshi Harashima
Journal of controlled release : official journal of the Controlled Release Society, 330, 753, 764, 10 Feb. 2021, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, This study reports on the development of a novel lipid combination that permits the efficient and highly selective delivery of plasmid DNA (pDNA) to immune cells in the spleen. Using DODAP, an ionizable lipid that was previously thought to be inefficient for gene delivery, we show for the first time, that this ignored lipid can be successfully used for efficient and targeted gene delivery in vivo, but only when combined with DOPE, a specific helper lipid. Using certain DODAP and DOPE ratios resulted in the formation of lipid nanoparticles (LNPs) with a ~ 1000-fold higher gene expression, and this expression was specific for the spleen, making it the most spleen-selective system for transfection using pDNA. The developed DODAP/DOPE-LNPs target immune cells in the spleen via receptors for complement C3 and this pathway is critical for efficient gene expression. We hypothesize that the high spleen transfection activity of DODAP/DOPE-LNPs is caused by the promotion of gene expression associated with B cell activation via complement receptors. LNPs encapsulating tumor-antigen encoding pDNA showed both prophylactic and therapeutic anti-tumor effects. The optimized LNPs resulted in the production of different cytokines and antigen-specific antibodies as well as exerting antigen-specific cytotoxic effects. This study revives the use of DODAP in gene delivery and highlights the importance of using appropriate lipid combinations for delivering genes to specific cells.

A novel dual-targeted rosiglitazone-loaded nanoparticle for the prevention of diet-induced obesity via the browning of white adipose tissue.
Ryu Hiradate, Ikramy A Khalil, Aya Matsuda, Mika Sasaki, Kyoko Hida, Hideyoshi Harashima
Journal of controlled release : official journal of the Controlled Release Society, 329, 665, 675, 10 Jan. 2021, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Adipose tissue in the body is classified as white adipose tissue (WAT); a fat-accumulating tissue, or brown adipose tissue (BAT); an energy-dissipating tissue. Transforming WAT-to-BAT (browning) is a promising strategy for the treatment of obesity, since it would lead to an increase in energy expenditure. Rosiglitazone (Rosi), an agonist of the peroxisome proliferator-activated receptor γ (PPARγ), is known to be a potent browning inducer in subcutaneous WAT. However, the effectiveness of Rosi has been quite limited because of several off-target effects. The objective of this study was to develop locally administered Rosi-loaded nanoparticles (Rs-NPs) with the ability to target adipocytes to achieve the adipose tissue-specific activation of PPARγ, thus causing the browning of WAT. We prepared dual targeted Rs-NPs that were modified with a specific peptide that targets prohibitin that are expressed in adipocytes, and a cell penetrating peptide for enhancing cellular uptake and controlling intracellular trafficking. The Rs-NPs modified with a single ligand were internalized into mature adipocytes and induced browning activity in vitro but they failed to significantly affect the body weight of the diet-induced obese mice model. The dual-targeted Rs-NPs induced a strong browning activity, both in vitro and in vivo, and successfully inhibited the progression of obesity, as evidenced by the shrinkage of hypertrophied adipocytes without any detectable systemic adverse effects. Meanwhile, free Rosi aggravated hepatic steatosis and did not cause adipose tissue browning nor the inhibition of body weight gain. We conclude that the increased energy expenditure via adipose tissue browning using dual-targeted Rs-NP is a promising strategy for the treatment of obesity and its related metabolic syndrome.

Engineered ε-decalactone lipomers bypass the liver to selectively in vivo deliver mRNA to the lungs without targeting ligands
Mahmoud M. Abd Elwakil, Tianle Gao, Takuya Isono, Yusuke Sato, Yaser H. A. Elewa, Toshifumi Satoh, Hideyoshi Harashima
Materials Horizons, Royal Society of Chemistry (RSC), 2021
Scientific journal,

Engineered lipomers coming from sustainable sources can efficiently bypass the liver to deliver a genetic message to the lungs after systemic administration without targeting ligands.

Three-dimensional, symmetrically assembled microfluidic device for lipid nanoparticle production
Niko Kimura, Masatoshi Maeki, Kosuke Sasaki, Yusuke Sato, Akihiko Ishida, Hirofumi Tani, Hideyoshi Harashima, Manabu Tokeshi
RSC ADVANCES, 11, 3, 1430, 1439, Jan. 2021
English, Scientific journal

Targeting the Mitochondrial Genome Via a MITO-Porter : Evaluation of mtDNA and mtRNA Levels and Mitochondrial Function.
Yuma Yamada, Hideyoshi Harashima
Methods in molecular biology (Clifton, N.J.), 2275, 227, 245, 2021, [Peer-reviewed], [Last author], [International Magazine]
English, Scientific journal, Genetic mutations and defects in mitochondrial DNA (mtDNA) are associated with certain types of mitochondrial dysfunctions, ultimately resulting in the emergence of a variety of human diseases. To achieve an effective mitochondrial gene therapy, it will be necessary to deliver therapeutic agents to the innermost mitochondrial space (the mitochondrial matrix), which contains the mtDNA pool. We recently developed a MITO-Porter, a liposome-based nanocarrier that delivers cargo to mitochondria via a membrane-fusion mechanism. In this chapter, we discuss the methodology used to deliver bioactive molecules to the mitochondrial matrix using a Dual Function (DF)-MITO-Porter, a liposome-based nanocarrier that delivers it cargo by means of a stepwise process, and an evaluation of mtDNA levels and mitochondrial activities in living cells. We also discuss mitochondrial gene silencing by the mitochondrial delivery of antisense RNA oligonucleotide (ASO) targeting mtDNA-encoded mRNA using the MITO-Porter system.

Current Status and Challenges Associated with CNS-Targeted Gene Delivery across the BBB.
Seigo Kimura, Hideyoshi Harashima
Pharmaceutics, 12, 12, 15 Dec. 2020, [Peer-reviewed], [Invited], [Last author], [International Magazine]
English, Scientific journal, The era of the aging society has arrived, and this is accompanied by an increase in the absolute numbers of patients with neurological disorders, such as Alzheimer's disease (AD) and Parkinson's disease (PD). Such neurological disorders are serious costly diseases that have a significant impact on society, both globally and socially. Gene therapy has great promise for the treatment of neurological disorders, but only a few gene therapy drugs are currently available. Delivery to the brain is the biggest hurdle in developing new drugs for the central nervous system (CNS) diseases and this is especially true in the case of gene delivery. Nanotechnologies such as viral and non-viral vectors allow efficient brain-targeted gene delivery systems to be created. The purpose of this review is to provide a comprehensive review of the current status of the development of successful drug delivery to the CNS for the treatment of CNS-related disorders especially by gene therapy. We mainly address three aspects of this situation: (1) blood-brain barrier (BBB) functions; (2) adeno-associated viral (AAV) vectors, currently the most advanced gene delivery vector; (3) non-viral brain targeting by non-invasive methods.

The use of design of experiments with multiple responses to determine optimal formulations for in vivo hepatic mRNA delivery.
Akari Hashiba, Manaya Toyooka, Yusuke Sato, Masatoshi Maeki, Manabu Tokeshi, Hideyoshi Harashima
Journal of controlled release : official journal of the Controlled Release Society, 327, 467, 476, 10 Nov. 2020, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Although great advances have been made in the delivery of short RNAs by lipid nanoparticles (LNPs), the optimal formulation composition and physicochemical properties of LNPs for long RNA (including mRNA) remain unclear. In the present study, we optimized the lipid composition of liver-targeted mRNA-loaded LNPs that were prepared with pH-sensitive cationic lipids that had been previously designed for siRNA delivery through a two stepped design of experiment (DoE). Multiple responses including physicochemical properties, gene expression, and liver-specificity were analyzed in order, not only to understand the role of each formulation parameter, but also to examine parameters that would be difficult to predict. We found that particle size and the PEG-to-phospholipid (PEG/PL) ratio were additional key factors for liver-specific gene expression in addition to the other formulation factors. The optimized formulation showed a better gene expression compared to other lipid formulations from industry leaders. These findings suggest that a "DoE with multiple responses" approach can be used to predict significant parameters and permit optimized formulations to be prepared more efficiently.

Evolution of drug delivery system from viewpoint of controlled intracellular trafficking and selective tissue targeting toward future nanomedicine.
Yuma Yamada, Yusuke Sato, Takashi Nakamura, Hideyoshi Harashima
Journal of controlled release : official journal of the Controlled Release Society, 327, 533, 545, 10 Nov. 2020, [Peer-reviewed], [Invited], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Due to the rapid changes that have occurred in the field of drug discovery and the recent developments in the early 21st century, the role of drug delivery systems (DDS) has become increasingly more important. For the past 20 years, our laboratory has been developing gene delivery systems based on lipid-based delivery systems. One of our efforts has been directed toward developing a multifunctional envelope-type nano device (MEND) by modifying the particle surface with octaarginine, which resulted in a remarkably enhanced cellular uptake and improved intracellular trafficking of plasmid DNA (pDNA). When we moved to in vivo applications, however, we were faced with the PEG-dilemma and we shifted our strategy to the incorporation of ionizable cationic lipids into our system. This resulted in some dramatic improvements over our original design and this can be attributed to the development of a new lipid library. We have also developed a mitochondrial targeting system based on a membrane fusion mechanism using a MITO-Porter, which can deliver nucleic acids/pDNA into the matrix of mitochondria. After the appearance of antibody medicines, Opdivo, an immune checkpoint inhibitor, has established cancer immunology as the 4th strategy in cancer therapy. Our DDS technologies can also be applied to this new field of cancer therapy to cure cancer by controlling our immune mechanisms. The latest studies are summarized in this review article.

ミトコンドリア病(Leigh脳症)モデルマウスの心機能の評価とモデルマウス由来心筋前駆細胞のミトコンドリア機能の検討　　　　　　　　　　　　　　　
佐々木 大輔, 武田 充人, 山田 勇磨, 原島 秀吉
日本小児循環器学会雑誌, 36, Suppl.2, s2, 358, (NPO)日本小児循環器学会, Nov. 2020
Japanese

The use of a MITO-Porter to deliver exogenous therapeutic RNA to a mitochondrial disease's cell with a A1555G mutation in the mitochondrial 12S rRNA gene results in an increase in mitochondrial respiratory activity.
Yuma Yamada, Minako Maruyama, Tomoko Kita, Shin-Ichi Usami, Shin-Ichiro Kitajiri, Hideyoshi Harashima
Mitochondrion, 55, 134, 144, Nov. 2020, [Peer-reviewed], [Last author], [International Magazine]
English, Scientific journal, We report on validating a mitochondrial gene therapeutic strategy using fibroblasts derived from patients with an A1555G point mutation in mitochondrial DNA coding 12S ribosomal RNA (rRNA (12S)). Wild-type rRNA (12S) as a therapeutic RNA was encapsulated in a mitochondrial targeting liposome, a MITO-Porter (rRNA-MITO-Porter), and an attempt was made to deliver the MITO-Porter to mitochondria of the diseased cells. It was confirmed that the rRNA-MITO-Porter treatment significantly decreased the ratio of the mutant rRNA content. Moreover, it was shown that the mitochondrial respiratory activities of the diseased cells were improved as the result of the mitochondrial transfection of the rRNA-MITO-Porter.

New Design Strategies for Controlling the Rate of Hydrophobic Drug Release from Nanoemulsions in Blood Circulation.
Saed Abbasi, Yusuke Sato, Kazuaki Kajimoto, Hideyoshi Harashima
Molecular pharmaceutics, 17, 10, 3773, 3782, 05 Oct. 2020, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, The intravenous administration of drug-loaded nanoparticles (NPs) is needed to achieve passive or active targeting in disease tissues. However, when the loaded drug is a hydrophobic small molecule, the NPs fail to reach adequate plasma drug concentrations mainly because of premature drug release. The pharmacokinetics of such drugs can be controlled by covalent modification, but this approach could compromise the safety or potency of the drug. In this study, we investigated two formulation parameters that could be used to improve the plasma concentrations of unmodified drugs that are loaded in a nanoemulsion (NE), a core-shell type NP. The first parameter is the loading ratio, and the second is the affinity of the drug to the core. Optimized NEs with reduced drug loading and with a high drug-core affinity resulted in a 12.4- and 11.2-fold increase in the plasma retention of curcumin and paclitaxel, respectively. Our strategy for enhancing the drug-core interaction affinity relied on mixing oils and surfactants to achieve cooperativity in noncovalent interactions, such as hydrophobic interactions, hydrogen bonding, and π-π stacking, which was further confirmed by theoretical calculations of interaction affinities. Finally, we report on the development of a cinnamic acid-derived oil-like material as a novel drug vehicle with exceptional solubilizing ability that could be used in intravenous formulations of NEs.

リポソームsiRNAデリバリーシステムを用いた腫瘍血管biglycanを標的とした新規血管新生阻害療法　　　　　　　　　　　　　　　
間石 奈湖, 櫻井 遊, 畠山 浩人, Li Cong, Alam Mohammad T., 菊地 央, 森本 浩史, 森本 真弘, 樋田 泰浩, 原島 秀吉, 樋田 京子
日本癌学会総会記事, 79回, OE16, 5, (一社)日本癌学会, Oct. 2020
English

核酸医薬とナノメディシン 多機能性エンベロープ型ナノ構造体の開発とナノ医療への展開　　　　　　　　　　　　　　　
原島 秀吉, 佐藤 悠介, 中村 孝司, 山田 勇磨
日本癌学会総会記事, 79回, ML13, ML13, (一社)日本癌学会, Oct. 2020
English

Manipulating the function of tumor-associated macrophages by siRNA-loaded lipid nanoparticles for cancer immunotherapy.
Nour Shobaki, Yusuke Sato, Yuichi Suzuki, Nana Okabe, Hideyoshi Harashima
Journal of controlled release : official journal of the Controlled Release Society, 325, 235, 248, 10 Sep. 2020, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, The tumor-microenvironment contains large numbers of tumor-associated macrophages (TAMs) which are largely M2 phenotypes and are involved in pro-tumorous functions. Targeting TAMs so as to manipulate them and to modify their functions could be a novel immunotherapy for the treatment of cancer. Such a strategy would involve targeting TAMs with short interfering RNA (siRNA) to modify their functions by silencing certain genes that are responsible for their M2 polarization. In this study, a lipid nanoparticle (LNP) formulation was used to target and deliver siRNA to TAMs. The LNP was mainly composed of a novel, pH-sensitive cationic lipid, referred to as the CL4H6 lipid, which had previously been optimized to target hepatocytes. The optimized siRNA-loaded CL4H6-LNPs were selectively and efficiently taken up and showed strong gene silencing activity in TAMs in a human tumor xenograft model in nude mice. Furthermore, an anti-tumor therapeutic response in the same tumor model was obtained by targeting TAMs using the optimized siRNA-loaded CL4H6-LNPs. The anti-tumor therapeutic response was obtained through the silencing of the signal transducer and activator of transcription 3 (STAT3) and hypoxia inducible factor 1 α (HIF-1α), which resulted in an increase in the level of infiltrated macrophage (CD11b+ cells) into the tumor-microenvironment (TME) as well as a tendency to increase the concentration of M1 macrophages (CD169+ cells). The treatment also resulted in reversing the pro-tumorous functions of TAMs -mainly angiogenesis and tumor cell activation-, as evidenced by a decrease in the related gene expression at the mRNA level. This research has promising clinical and pharmaceutical applications for achieving novel macrophage-based cancer immunotherapy.

Challenges in Promoting Mitochondrial Transplantation Therapy.
Yuma Yamada, Momo Ito, Manae Arai, Mitsue Hibino, Takao Tsujioka, Hideyoshi Harashima
International journal of molecular sciences, 21, 17, 02 Sep. 2020, [Peer-reviewed], [Last author], [International Magazine]
English, Scientific journal, Mitochondrial transplantation therapy is an innovative strategy for the treatment of mitochondrial dysfunction. The approach has been reported to be useful in the treatment of cardiac ischemic reperfusion injuries in human clinical trials and has also been shown to be useful in animal studies as a method for treating mitochondrial dysfunction in various tissues, including the heart, liver, lungs, and brain. On the other hand, there is no methodology for using preserved mitochondria. Research into the pharmaceutical formulation of mitochondria to promote mitochondrial transplantation therapy as the next step in treating many patients is urgently needed. In this review, we overview previous studies on the therapeutic effects of mitochondrial transplantation. We also discuss studies related to immune responses that occur during mitochondrial transplantation and methods for preserving mitochondria, which are key to their stability as medicines. Finally, we describe research related to mitochondrial targeting drug delivery systems (DDS) and discuss future perspectives of mitochondrial transplantation.

ミトコンドリアDDSを用いたビタミンB1の脳内送達・治療効果の検証　　　　　　　　　　　　　　　
山田 勇磨, 石丸 拓也, 原島 秀吉
日本DDS学会学術集会プログラム予稿集, 36回, 169, 169, 日本DDS学会, Aug. 2020
Japanese

Mitochondrial Delivery of an Anticancer Drug Via Systemic Administration Using a Mitochondrial Delivery System That Inhibits the Growth of Drug-Resistant Cancer Engrafted on Mice.
Yuma Yamada, Reina Munechika, Satrialdi, Fumika Kubota, Yusuke Sato, Yu Sakurai, Hideyoshi Harashima
Journal of pharmaceutical sciences, 109, 8, 2493, 2500, Aug. 2020, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Mitochondrial delivery of an anticancer drug targeting cancer cells would eventually result in cell death. To achieve this, a drug delivery system targeting mitochondria is needed. We recently developed a MITO-Porter, a liposome that delivers its cargo to mitochondria. We reported that such a MITO-Porter could deliver doxorubicin (DOX), an anticancer drug, to mitochondria in OS-RC-2 cells, a drug resistant cancer cell, resulting in inhibiting the cell growth, based in in vitro experiments. Herein, we report on validating the benefit of such a therapeutic strategy for treating drug resistant cancers by the in vivo targeting of mitochondria. We prepared a DOX-MITO-Porter, in which DOX was encapsulated in the MITO-Porter and optimized its retention in blood circulation. When the DOX-MITO-Porter was administered to mice bearing OS-RC-2 cells via tail vein injection, tumor size was significantly decreased, compared to DOX itself and to the DOX-encapsulated polyethylene glycol-modified liposome (DOX-PEG-LP). Intracellular observation confirmed that the DOX-MITO-Porter had accumulated in tumor mitochondria. It was also found a relationship between anti-tumor effect and the mitochondrial function, as indicated by the depolarization of mitochondrial membrane potential. This study provides support for the utility of an in vivo mitochondrial delivery system in drug resistant cancer therapies.

Development of a Microfluidic-Based Post-Treatment Process for Size-Controlled Lipid Nanoparticles and Application to siRNA Delivery.
Niko Kimura, Masatoshi Maeki, Yusuke Sato, Akihiko Ishida, Hirofumi Tani, Hideyoshi Harashima, Manabu Tokeshi
ACS applied materials & interfaces, 12, 30, 34011, 34020, 29 Jul. 2020, [Peer-reviewed], [International Magazine]
English, Scientific journal, Microfluidic methodologies for preparation of lipid nanoparticles (LNPs) based on an organic solvent injection method enable precise size control of the LNPs. After preparation of LNPs, the organic solvent injection method needs some post-treatments, such as overnight dialysis or direct dilution with a buffer solution. LNP production using the microfluidic-based organic solvent injection method is dominated by kinetics rather than thermodynamics. Kinetics of ethanol removal from the inner and outer membranes of LNPs could induce a structural change in the membrane that could lead to fusion of LNPs. However, the effects of microfluidic post-treatment on the final size of LNPs have not been sufficiently understood. Herein, we investigated the effect of the post-treatment processes on the final product size of LNPs in detail. A simple baffle device and a model lipid system composed of a neutral phospholipid (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, POPC) and cholesterol were used to produce the LNPs. We demonstrated that flow conditions of the post-treatment diluting the remaining ethanol in the LNP suspension affected the final product size of LNPs. Based on the findings, we developed an integrated baffle device composed of an LNP production region and a post-treatment region for a microfluidic-based LNP production system; this integrated baffle device prevented the undesirable aggregation or fusion of POPC LNPs even for the high-lipid-concentration condition. Finally, we applied our concept to small interfering RNA (siRNA) delivery and confirmed that no significant effects due to the continuous process occurred on the siRNA encapsulation efficiency, biological distribution, and knockdown activity. The microfluidic post-treatment method is expected to contribute to the production of LNPs for practical applications and the development of novel LNP-based nanomedicines.

Lipid nanoparticles fuse with cell membranes of immune cells at low temperatures leading to the loss of transfection activity.
Takashi Nakamura, Koharu Yamada, Yusuke Sato, Hideyoshi Harashima
International journal of pharmaceutics, 587, 119652, 119652, 17 Jul. 2020, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Delivering nucleic acid using a non-viral vector is a potent strategy for gene modification and controlling gene expression in immune cell therapy. Since the low-temperature storage (0-4 °C) or cryopreservation of cells are indispensable for performing immune cell therapy, we investigated the interactions between an siRNA-loaded lipid nanoparticle (LNP), a multifunctional envelope-type nanodevice (MEND) containing YSK12-C4 (YSK12-MEND), and human immune cell lines (NK-92 and Jurkat) at low-temperature and its effect on transfection activity. The YSK12-MEND readily bound to the cell membrane of NK-92 cells at low-temperature, but no internalization of the YSK12-MEND by cells was observed, even after returning the temperature to 37 °C. Gene silencing activity was completely impaired. The cause of this inhibition appears to be membrane fusion between the YSK12-MEND and cell membrane at the low-temperature. Collectively, our results suggest that the exposure of siRNA-loaded LNPs to cells at low-temperature should be avoided in defining transfection protocols in immune cell therapy.

Different kinetics for the hepatic uptake of lipid nanoparticles between the apolipoprotein E/low density lipoprotein receptor and the N-acetyl-D-galactosamine/asialoglycoprotein receptor pathway
Yusuke Sato, Yoshiyuki Kinami, Kazuki Hashiba, Hideyoshi Harashima
Journal of Controlled Release, 322, 217, 226, Elsevier B.V., 10 Jun. 2020
English, Scientific journal

Erratum: The optimization of cancer photodynamic therapy by utilization of a pi-extended porphyrin-type photosensitizer in combination with MITO-Porter (Chem. Commun. (2020) 56 (1145–1148) DOI: 10.1039/C9CC08563G)
Satrialdi, Reina Munechika, Vasudevanpillai Biju, Yuta Takano, Hideyoshi Harashima, Yuma Yamada
Chemical Communications, 56, 45, 6153, Royal Society of Chemistry, 07 Jun. 2020
English, Scientific journal

Dawn of lipid nanoparticles in lymph node targeting: Potential in cancer immunotherapy.
Takashi Nakamura, Hideyoshi Harashima
Advanced drug delivery reviews, 167, 78, 88, 06 Jun. 2020, [Peer-reviewed], [Invited], [Last author], [International Magazine]
English, Scientific journal, It is generally known that the lymph nodes (LNs) are important tissues in cancer immunotherapy. Therefore, delivering immune functional compounds to LNs is a useful strategy for enhancing cancer immunotherapy. Lipid-based nanocarriers have been widely used as delivery systems that target LNs, but lipid nanoparticle (LNP) technology has recently attracted increased interest. High levels of nucleic acids can be efficiently loaded in LNPs, they can be used to actively deliver nucleic acids into the cytoplasm, and they can be produced on an industrial scale. The use of microfluidic devices has been particularly valuable for producing small-sized LNPs, thus paving the way for successful LN targeting. In the review, we focus on the potential of LNP technology for targeting LNs.

Validation of Gene Therapy for Mutant Mitochondria by Delivering Mitochondrial RNA Using a MITO-Porter.
Eriko Kawamura, Minako Maruyama, Jiro Abe, Akira Sudo, Atsuhito Takeda, Shingo Takada, Takashi Yokota, Shintaro Kinugawa, Hideyoshi Harashima, Yuma Yamada
Molecular therapy. Nucleic acids, 20, 687, 698, 05 Jun. 2020, [Peer-reviewed], [International Magazine]
English, Scientific journal, Here, we report on validating a mitochondrial gene therapy by delivering nucleic acids to mitochondria of diseased cells by a MITO-Porter, a liposome-based carrier for mitochondrial delivery. We used cells derived from a patient with a mitochondrial disease with a G625A heteroplasmic mutation in the tRNAPhe of the mitochondrial DNA (mtDNA). It has been reported that some mitochondrial gene diseases are caused by heteroplasmic mutations, in which both mutated and wild-type (WT) genes are present, and the accumulation of pathological mutations leads to serious, intractable, multi-organ diseases. Therefore, the decrease of the mutated gene rate is considered to be a useful gene therapy strategy. To accomplish this, wild-type mitochondrial pre-tRNAPhe (pre-WT-tRNAPhe), prepared by in vitro transcription, was encapsulated in the MITO-Porter. The pre-WT-tRNAPhe encapsulated in the MITO-Porter was transfected into diseased mitochondrial cells, and the resulting mutant levels were examined by an amplification refractory mutation system (ARMS)-quantitative PCR. The mutation rate of tRNAPhe was decreased, and this therapeutic effect was sustained even on the 8th day after transfection. Furthermore, mitochondrial respiratory activity of the disease cells was increased after the transfection of therapeutic pre-WT-tRNAPhe. These results support the conclusion that the mitochondrial delivery of therapeutic nucleic acids represents a viable strategy for mitochondrial gene therapy.

Distribution of BCG-CWS-Loaded Nanoparticles in the Spleen After Intravenous Injection Affects Cytotoxic T Lymphocyte Activity.
Hideyuki Masuda, Takashi Nakamura, Hideyoshi Harashima
Journal of pharmaceutical sciences, 109, 6, 1943, 1950, Jun. 2020, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Interest has developed in the bacillus Calmette-Guerin (BCG) cell wall skeleton (BCG-CWS) as a noninfectious adjuvant. Although BCG-CWS readily undergoes aggregation, in a previous study, we applied it to cancer immunotherapy via intravenous administration by encapsulating the BCG-CWS into nanoparticles (CWS-NPs). The CWS-NPs were taken up by major histocompatibility complex (MHC) class II+ (MHC-II+) cells and induced antigen-specific cytotoxic T lymphocyte (CTL) activity. However, the nature of the contribution of MHC-II+ cells to the CTL response continues to be unclear. In this study, we investigated the relationship between the distribution of CWS-NPs in the spleen and CTL activity. The main MHC-II+ cells that internalized the CWS-NPs were B cells. Decreasing the level of polyethylene glycol modification increased the uptake of CWS-NPs by B cells, leading to an increased CTL activity. A comparison of CWS-NPs with different uptake efficiencies into dendritic cells and B cells suggested that the DCs with internalized CWS-NPs may contribute to CTL activation compared with B cells. We succeeded in enhancing CTL activity by the CWS-NPs, and the findings reported herein should provide important information regarding target cells for the development of CWS-NP.

Validation of a mitochondrial RNA therapeutic strategy using fibroblasts from a Leigh syndrome patient with a mutation in the mitochondrial ND3 gene.
Yuma Yamada, Kana Somiya, Akihiko Miyauchi, Hitoshi Osaka, Hideyoshi Harashima
Scientific reports, 10, 1, 7511, 7511, 05 May 2020, [International Magazine]
English, Scientific journal, We report on the validation of a mitochondrial gene therapeutic strategy using fibroblasts from a Leigh syndrome patient by the mitochondrial delivery of therapeutic mRNA. The treatment involves delivering normal ND3 protein-encoding mRNA as a therapeutic RNA to mitochondria of the fibroblasts from a patient with a T10158C mutation in the mtDNA coding the ND3 protein, a component of the mitochondrial respiratory chain complex I. The treatment involved the use of a liposome-based carrier (a MITO-Porter) for delivering therapeutic RNA to mitochondria via membrane fusion. The results confirmed that the mitochondrial transfection of therapeutic RNA by the MITO-Porter system resulted in a decrease in the levels of mutant RNA in mitochondria of diseased cells based on reverse transcription quantitative PCR. An evaluation of mitochondrial respiratory activity by respirometry also showed that transfection using the MITO-Porter resulted in an increase in maximal mitochondrial respiratory activity in the diseased cells.

脂質ナノ粒子の特性がリンパ節送達とリンパ節内分布へ与える影響　　　　　　　　　　　　　　　
中村 孝司, 河合 美典, 佐藤 悠介, 真栄城 正寿, 渡慶次 学, 原島 秀吉
日本薬剤学会年会講演要旨集, 35年会, 171, 171, (公社)日本薬剤学会, May 2020
Japanese

MELAS A3243G変異型ミトコンドリアDNAを標的とした遺伝子治療戦略の検証　　　　　　　　　　　　　　　
山田 勇磨, 宗宮 加奈, 佐々木 大輔, 武田 充人, 原島 秀吉
日本薬剤学会年会講演要旨集, 35年会, 169, 169, (公社)日本薬剤学会, May 2020
Japanese

Involvement of Caveolin-1-mediated transcytosis in the intratumoral accumulation of liposomes.
Yu Sakurai, Akari Kato, Hideyoshi Harashima
Biochemical and biophysical research communications, 525, 2, 313, 318, 30 Apr. 2020, [International Magazine]
English, Scientific journal, For achieving efficient cancer treatment, it is important to elucidate the mechanism responsible for the accumulation of nanoparticles in tumor tissue. Recent studies suggest that nanoparticles are not delivered merely through gaps between tumor endothelial cells. We previously reported that the maturation of the vascular structure by the vascular endothelial cell growth factor receptor 2 (VEGFR2) using a previously developed siRNA delivery technology (RGD-MEND) significantly enhanced the accumulation of nanoparticles in types of cancers that area vessel-rich (renal cell carcinoma). This result was completely inconsistent with the generally accepted theory of the enhanced permeability and retention (EPR) effect. We hypothesized that a caveolin-1 (Cav1)-mediated transcellular route would be involved with the penetration of nanoparticles into tumor vasculature. To reveal the exact mechanism responsible for this enhancement, we observed the delivery of long-circulating liposomes (LPs) after Cav1 was co-suppressed by RGD-MEND with VEGFR2. The enhanced delivery of LPs by siRNA against VEGFR2 (siVEGFR2) was accompanied by the elevated expression of the Cav1 protein. In addition, Cav1 knockdown by siRNA against Cav1 (siCav1) canceled the enhanced delivery of LPs by siVEGFR2. The injection of siCav1 had no effect on the formation of alpha smooth muscle actin or vascular endothelial cell adhesion molecules. These results suggest that a Cav1-induced transcellular route and not a paracellular route, at least partially, contributes to the accumulation of nanoparticles in tumors.

Improved Stability of siRNA-Loaded Lipid Nanoparticles Prepared with a PEG-Monoacyl Fatty Acid Facilitates Ligand-Mediated siRNA Delivery.
Yu Sakurai, Wataru Mizumura, Kenichiro Ito, Kazuhiro Iwasaki, Takayuki Katoh, Yuki Goto, Hiroaki Suga, Hideyoshi Harashima
Molecular pharmaceutics, 17, 4, 1397, 1404, 06 Apr. 2020, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Peptide modification is a popular strategy for developing an active targeting lipid nanoparticle (LNP). In modifying the surface of an LNP with a peptide, the sequence and structure of the peptide strongly affects the formation of the LNP. Specifically, a peptide with a high hydrophobicity can induce coarsening and aggregation of the LNP. In an attempt to prevent this from occurring, we incorporated monoacyl and diacyl group-conjugated poly(ethylene glycol) (PEG) into a LNP. We previously developed an original LNP, a multifunctional envelope type nanodevice (MEND) modified with an Epi-1 peptide, a ligand with a high affinity for the epithelial cell adhesion molecule (EpCAM). Using this peptide-modified MEND, the efficiency of delivery of a small interfering RNA (siRNA) encapsulated in the MEND was significantly improved. Although increasing the ratio of modification enhanced cellular uptake, the increase also induced aggregation of the LNP, particularly in the case of a large scale preparation. Our results indicate that a monoacyl PEG-lipid can prevent aggregation, even when the LNP is modified with higher molar ratios of peptide, but that this also results in a decrease in delivery efficiency. Moreover, the Epi-1-modified MEND exhibited a strong silencing effect in an ovarian cancer peritoneal dissemination model. Our results suggest that the simple incorporation of a monoacyl derivative into the PEG-lipid resulted in the formation of a peptide-modified LNP with improved characteristics.

Vitamin E Scaffolds of pH-Responsive Lipid Nanoparticles as DNA Vaccines in Cancer and Protozoan Infection.
Mio Maeta, Naoya Miura, Hiroki Tanaka, Takashi Nakamura, Ryo Kawanishi, Yoshifumi Nishikawa, Kenichi Asano, Masato Tanaka, Shinya Tamagawa, Yuta Nakai, Kota Tange, Hiroki Yoshioka, Hideyoshi Harashima, Hidetaka Akita
Molecular pharmaceutics, 17, 4, 1237, 1247, 06 Apr. 2020, [Peer-reviewed], [International Magazine]
English, Scientific journal, DNA vaccinations are promising strategies for treating diseases that require cellular immunity (i.e., cancer and protozoan infection). Here, we report on the use of a liposomal nanocarrier (lipid nanoparticles (LNPs)) composed of an SS-cleavable and pH-activated lipidlike material (ssPalm) as an in vivo DNA vaccine. After subcutaneous administration, the LNPs containing an ssPalmE, an ssPalm with vitamin E scaffolds, elicited a higher gene expression activity in comparison with the other LNPs composed of the ssPalms with different hydrophobic scaffolds. Immunization with the ssPalmE-LNPs encapsulating plasmid DNA that encodes ovalbumin (OVA, a model tumor antigen) or profilin (TgPF, a potent antigen of Toxoplasma gondii) induced substantial antitumor or antiprotozoan effects, respectively. Flow cytometry analysis of the cells that had taken up the LNPs in draining lymph nodes (dLNs) showed that the ssPalmE-LNPs were largely taken up by macrophages and a small number of dendritic cells. We found that the transient deletion of CD169+ macrophages, a subpopulation of macrophages that play a key role in cancer immunity, unexpectedly enhanced the activity of the DNA vaccine. These data suggest that the ssPalmE-LNPs are effective DNA vaccine carriers, and a strategy for avoiding their being trapped by CD169+ macrophages will be a promising approach for developing next-generation DNA vaccines.

Lipid Nanoparticles for Cell-Specific in Vivo Targeted Delivery of Nucleic Acids
Ikramy A. Khalil, Mahmoud A. Younis, Seigo Kimura, Hideyoshi Harashima
Biological and Pharmaceutical Bulletin, 43, 4, 584, 595, Pharmaceutical Society of Japan, 01 Apr. 2020, [Peer-reviewed]
English, Scientific journal

The Effect of Size and Charge of Lipid Nanoparticles Prepared by Microfluidic Mixing on Their Lymph Node Transitivity and Distribution.
Takashi Nakamura, Minori Kawai, Yusuke Sato, Masatoshi Maeki, Manabu Tokeshi, Hideyoshi Harashima
Molecular pharmaceutics, 17, 3, 944, 953, 02 Mar. 2020, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Because the lymph node (LN) is a critical organ for inducing immune responses against pathogens and cancers, the transport of immune functional molecules such as antigens and adjuvants to LNs by delivery systems is a useful strategy for the effective outcome of an immune response. The size and charge of a delivery system largely affect the transitivity to and distribution within LN. Although pH-sensitive lipid nanoparticles (LNPs) prepared by microfluidic mixing are the latest delivery system to be applied clinically, the effects of their size and charge on the transitivity to and distribution within LN are currently unknown. We investigated the size and charge effect of LNPs prepared by microfluidic mixing on transitivity to and distribution within LNs. A 30 nm-sized LNP (30-LNP) was efficiently translocated to LNs and was taken up by CD8+ dendritic cells, while the efficiency was drastically decreased in the cases of 100 and 200 nm-sized LNPs. Furthermore, a comparative study between neutral, positively, and negatively charged 30-LNP revealed that the negative 30-LNP moved to the LN more efficiently than the other LNPs. Interestingly, the negative 30-LNP reached the deep cortex, namely, the T cell zone. Our findings provide informative insights for designing LN-targeting LNPs prepared by microfluidic mixing and for the translocation of nanoparticles in LNs.

The gala peptide: multiple roles in drug and gene delivery
Ikramy A. Khalil, Hideyoshi Harashima
Bulletin of Pharmaceutical Sciences, 43, 1, 39, 51, Assiut University, 01 Mar. 2020
English, Scientific journal

光増感分子rTPAを搭載したミトコンドリア標的型ナノカプセルの構築および担癌モデルマウスを用いた癌光治療戦略の検証　　　　　　　　　　　　　　　
山田 勇磨, Satrialdi, 高野 勇太, Biju Vasudevanpillai, 原島 秀吉
日本薬学会年会要旨集, 140年会, 26X, am11, (公社)日本薬学会, Mar. 2020
Japanese

Development of a nanoparticle that releases nucleic acids in response to a mitochondrial environment.
Yuma Yamada, Yutaka Fukuda, Daisuke Sasaki, Minako Maruyama, Hideyoshi Harashima
Mitochondrion, 52, 67, 74, 22 Feb. 2020, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, The delivery of nucleic acids targeting mutant mtDNA represent a potential strategy for addressing a variety of mitochondria-related diseases. We previously developed a MITO-Porter, a nano carrier that is capable of delivering nanoparticles of nucleic acids to mitochondria of human cells. Here, we report on an investigation of a series of nanoparticles formed with various poly cationic peptides that can release nucleic acids in response to a mitochondrial environment. A significant relationship was found between the number of and the location of arginine and histidine residues in the peptide sequence and the release of nucleic acids in a mitochondrial environment.

The optimization of cancer photodynamic therapy by utilization of a pi-extended porphyrin-type photosensitizer in combination with MITO-Porter.
Satrialdi, Reina Munechika, Vasudevanpillai Biju, Yuta Takano, Hideyoshi Harashima, Yuma Yamada
Chemical communications (Cambridge, England), 56, 7, 1145, 1148, 23 Jan. 2020, [Peer-reviewed], [International Magazine]
English, Scientific journal, The uncontrolled production of reactive oxygen species during photodynamic therapy (PDT) induces oxidative stress. The full potential of PDT is accomplished by delivery of a pi-extended porphyrin-type photosensitizer into mitochondria of tumor cells using a MITO-Porter, a mitochondrial targeting nanodevice. This strategy can be implemented for innovative cancer therapy.

Therapeutic Strategies for Regulating Mitochondrial Oxidative Stress.
Yuma Yamada, Yuta Takano, Satrialdi, Jiro Abe, Mitsue Hibino, Hideyoshi Harashima
Biomolecules, 10, 1, 05 Jan. 2020, [Peer-reviewed], [Last author], [International Magazine]
English, Scientific journal, There have been many reports on the relationship between mitochondrial oxidative stress and various types of diseases. This review covers mitochondrial targeting photodynamic therapy and photothermal therapy as a therapeutic strategy for inducing mitochondrial oxidative stress. We also discuss other mitochondrial targeting phototherapeutic methods. In addition, we discuss anti-oxidant therapy by a mitochondrial drug delivery system (DDS) as a therapeutic strategy for suppressing oxidative stress. We also describe cell therapy for reducing oxidative stress in mitochondria. Finally, we discuss the possibilities and problems associated with clinical applications of mitochondrial DDS to regulate mitochondrial oxidative stress.

A small-sized lipid nanoparticles production method using microfluidic devices with baffle structures
Niko Kimura, Masatoshi Maeki, Yusuke Note, Yusuke Sato, Akihiko Ishida, Hirofumi Tani, Hideyoshi Harashima, Manabu Tokeshi
21st International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2017, 965, 966, 2020
International conference proceedings

Power of mitochondrial drug delivery systems to produce innovative nanomedicines.
Yuma Yamada, Satrialdi, Mitsue Hibino, Daisuke Sasaki, Jiro Abe, Hideyoshi Harashima
Advanced drug delivery reviews, 154-155, 187, 209, 2020, [Peer-reviewed], [Invited], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Mitochondria carry out various essential functions including ATP production, the regulation of apoptosis and possess their own genome (mtDNA). Delivering target molecules to this organelle, it would make it possible to control the functions of cells and living organisms and would allow us to develop a better understanding of life. Given the fact that mitochondrial dysfunction has been implicated in a variety of human disorders, delivering therapeutic molecules to mitochondria for the treatment of these diseases is an important issue. To date, several mitochondrial drug delivery system (DDS) developments have been reported, but a generalized DDS leading to therapy that exclusively targets mitochondria has not been established. This review focuses on mitochondria-targeted therapeutic strategies including antioxidant therapy, cancer therapy, mitochondrial gene therapy and cell transplantation therapy based on mitochondrial DDS. A particular focus is on nanocarriers for mitochondrial delivery with the goal of achieving mitochondria-targeting therapy. We hope that this review will stimulate the accelerated development of mitochondrial DDS.

Editorial: Advanced Liposome Research.
Hideyoshi Harashima, Tatsuhiro Ishida
Advanced drug delivery reviews, 154-155, 1, 1, 2020, [Invited], [Lead author, Corresponding author], [International Magazine]
English, Scientific journal

Hydrophobic scaffolds of pH-sensitive cationic lipids contribute to miscibility with phospholipids and improve the efficiency of delivering short interfering RNA by small-sized lipid nanoparticles
Yusuke Sato, Nana Okabe, Yusuke Note, Kazuki Hashiba, Masatoshi Maeki, Manabu Tokeshi, Hideyoshi Harashima
Acta Biomaterialia, 102, 341, 350, Jan. 2020, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Spleen selective enhancement of transfection activities of plasmid DNA driven by octaarginine and an ionizable lipid and its implications for cancer immunization
Seigo Kimura, Ikramy A. Khalil, Yaser H.A. Elewa, Hideyoshi Harashima
Journal of Controlled Release, 313, 70, 79, Elsevier BV, Nov. 2019
Scientific journal

Targeting Tumor Endothelial Cells with Nanoparticles.
Sakurai Y, Akita H, Harashima H
International journal of molecular sciences, 20, 23, Nov. 2019, [Peer-reviewed], [Last author, Corresponding author]

Innovative nanotechnologies for enhancing nucleic acids/gene therapy: Controlling intracellular trafficking to targeted biodistribution.
Nakamura T, Yamada Y, Sato Y, Khalil IA, Harashima H
Biomaterials, 218, 119329, 119329, Oct. 2019, [Peer-reviewed], [Invited], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Nanomedicine promises to play an important role in next generation therapy, including Nucleic acid/Gene therapy. To accomplish this, innovative nanotechnologies will be needed to support nanomedicine by controlling not only the biodistribution but also the intracellular trafficking of macromolecules such as RNA/DNA. A multifunctional envelope-type nano device (MEND) was developed to meet this requirement. We herein provide an update regarding the functions of the MEND system focusing on the introduction of different functional biomaterials that enhance efficiency. The octaarginine (R8) peptide enhances cellular uptake and controls intracellular trafficking to induce synergism in transgene expression. The R8 was also used for developing a MITO-Porter system for mitochondrial targeting. The function of the MITO-Porter system was extended by developing a mitochondrial reporter gene for mitochondrial gene therapy. For efficient in vivo gene delivery, new pH-sensitive lipids have been introduced to achieve controlled biodistribution and to enhance endosomal escape. For example, the CL4H6 lipid exerts a more efficient in vivo gene silencing than that of ONPATTROTM, a preparation that has been approved by the US Food and Drug Administration. We further summarize new technologies that have been successfully applied to cancer immunotherapy leading to the introduction of a new strategy based on the concept of the Cancer-Immunity Cycle.

A nanocarrier for the mitochondrial delivery of nucleic acids to cardiomyocytes.
Yamada Y, Fujishita N, Harashima H
Nucleosides, nucleotides & nucleic acids, 1, 15, Oct. 2019, [Peer-reviewed], [Last author]

Synergistic Enhancement of Cellular Uptake With CD44-Expressing Malignant Pleural Mesothelioma by Combining Cationic Liposome and Hyaluronic Acid-Lipid Conjugate.
Yu Sakurai, Akari Kato, Yasuhiro Hida, Junichi Hamada, Nako Maishi, Kyoko Hida, Hideyoshi Harashima
Journal of pharmaceutical sciences, 108, 10, 3218, 3224, Oct. 2019, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Malignant pleural mesothelioma (MPM) is a highly aggressive form of cancer, with a median survival of less than 1 year. It is well known that the hyaluronan (HA) receptor CD44 is highly expressed by MPM cells and is reported to be correlated with a poor prognosis. We herein report on the development of a new type if drug delivery system against CD44 that involves the use of lipid nanoparticles (LNPs) equipped with a new type of HA derivative. In this study, we evaluated HA-lipid conjugation (HAL) via the end of the HA molecule through reductive amination, a process that allowed the carboxylate group to remain intact. As a result, the HAL-modified LNP appears to be a potent nanoparticle for dealing with MPM. Surprisingly, the use of a combination of a cationic lipid and HAL had a synergistic effect on cellular uptake in MPM and consequently permitted an anti-cancer drug such as cis-diamminedichloro-platinum(II) (CDDP). Intrapleural injection of CDDP-loaded HAL-LNP (1.5 mg/kg as CDDP) per week significantly suppressed the progression of this type of cancer in an MPM orthotopic model. These results suggest that HAL-modified LNP represents a potent delivery system for MPM cells that express high levels of CD44.

A Multifunctional Lipid-Based Nanodevice for the Highly Specific Codelivery of Sorafenib and Midkine siRNA to Hepatic Cancer Cells
Mahmoud A. Younis, Ikramy A. Khalil, Mahmoud M. Abd Elwakil, Hideyoshi Harashima
Molecular Pharmaceutics, 16, 9, 4031, 4044, 03 Sep. 2019
Scientific journal

Hyaluronan-modified nanoparticles for tumor-targeting.
Yu Sakurai, Hideyoshi Harashima
Expert opinion on drug delivery, 16, 9, 915, 936, Sep. 2019, [International Magazine]
English, Scientific journal, Introduction: Hyaluronan (HA), a natural polysaccharide, is produced in large amounts by the human body. Since its receptor CD44 is highly expressed in many types of cancers, HA is a promising ligand for cancer-targeting nanoparticles (NPs). Since the enhanced permeability and retention (EPR) effect-based strategy faces difficulties in terms of efficiency in clinical studies, studies focusing on HA-modified NPs that can actively target cancer cells should be prominent for further progress in NP-based cancer treatment. Areas covered: Various materials combined with HA composed of lipid nanoparticles and polymers as well as iron, gold and other metals have been examined. In addition, their cargos have been quite diverse and include small molecule drugs, imaging agents, proteins and nucleic acids. We summarize recent studies on varieties of NPs and describe the properties of HA as a ligand of CD44. Expert opinion: In spite of a number of studies on HA-based NPs, there is few examples of HA-based NPs in clinical. In order to make a progress in clinical study, an evaluation methodology of HA-NPs should be unified and normalized.

The silencing of indoleamine 2,3-dioxygenase 1 (IDO1) in dendritic cells by siRNA-loaded lipid nanoparticles enhances cell-based cancer immunotherapy.
Endo R, Nakamura T, Kawakami K, Sato Y, Harashima H
Scientific reports, 9, 1, 11335, 11335, Aug. 2019, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Cell-based therapy using dendritic cells (DC) represents a potent cancer immunotherapy. However, activated DC express indoleamine 2,3-dioxygenase 1 (IDO1), a counter-regulatory and tolerogenic molecule, leading to the inhibition of T cell activation and the promotion of T cell differentiation into regulatory T cells. Silencing the IDO1 gene in DC by small interfering RNA (siRNA) represents a potent therapeutic strategy. We report on the successful and efficient introduction of a siRNA targeting IDO1 into mouse DCs by a means of a multifunctional envelope-type nanodevice (MEND) containing a YSK12-C4 (YSK12-MEND). The YSK12-C4 has both fusogenic and cationic properties. The YSK12-MEND induced an effective level of gene silencing of IDO1 at siRNA doses in the range of 1-20 nM, a concentration that commercially available transfection reagents are not able to silence. The YSK12-MEND mediated IDO1 silencing had no effect on the characteristic determinants of DC phenotype such as CD11c, CD80 and MHC class II. The silencing of IDO1 in DC by the YSK12-MEND significantly enhanced the antitumor effect against E.G7-OVA tumor. Moreover, a decrease in the numbers of regulatory T cells in the tumor was observed in mice that were treated with the IDO1-silenced DC. The YSK12-MEND appears to be a potent delivery system for IDO1-silenced DC based cancer immunotherapy.

Targeted mitochondrial delivery of antisense RNA-containing nanoparticles by a MITO-Porter for safe and efficient mitochondrial gene silencing.
Kawamura E, Hibino M, Harashima H, Yamada Y
Mitochondrion, 49, 178, 188, Aug. 2019, [Peer-reviewed], [International Magazine]
English, Scientific journal, Mitochondrial gene therapy will be needed to treat mitochondrial diseases. We previously demonstrated mitochondrial gene silencing by the mitochondrial delivery of antisense RNA oligonucleotide (ASO) targeting mtDNA-encoded mRNA using a MITO-Porter, a liposomal nano carrier system designed for mitochondrial delivery. Here, we report on the efficient packaging of ASO in the MITO-Porter via a nanoparticle packaging method, which showed a 10-fold higher packaging efficiency than the conventional method. The constructed carrier showed a decrease in the target mRNA levels and ATP production. These results indicate that such a MITO-Porter has potential for use in therapies designed to regulate mitochondrial function.

A study of the endocytosis mechanism and transendothelial activity of lung-targeted GALA-modified liposomes.
Santiwarangkool S, Akita H, Khalil IA, Abd Elwakil MM, Sato Y, Kusumoto K, Harashima H
Journal of controlled release : official journal of the Controlled Release Society, 307, 55, 63, Aug. 2019, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, The GALA peptide (WEAALAEALAEALAEHLAEALAEALEALAA) was originally designed to induce the destabilization of endosomal membranes based on its ability to undergo a pH-dependent conformational change from a random coil to an α-helix. We recently found that liposomes modified with GALA peptide (GALA-LPs) extensively accumulate in lung endothelial cells (ECs) after intravenous injection. However, the uptake mechanism of GALA-LPs and their ability to reach alveolar epithelium was unclear. We report herein that GALA-LPs are internalized into ECs via a clathrin-mediated pathway. Surprisingly, GALA-LPs had the ability to pass lung ECs and reach other cells through transcytosis. GALA-LPs were taken up by >70% of lung ECs, while they also accumulated in ~30% of type I alveolar epithelium (ATI). GALA-modified gold nanoparticles were detected in ECs, in the basement membrane and in other cells such as ATI, type II alveolar epithelium (ATII) and alveolar macrophages. Consistent with this result, a significant gene knockdown was achieved in lung epithelium cells using GALA-LPs encapsulating anti-podoplanin siRNA. This indicates that GALA-LPs can be used as a carrier for delivering macromolecules to parenchymal as well as to endothelial cells in the lung. Although caveolae are commonly linked to the transendothelial transport of proteins and antibodies, our data indicate that clathrin-mediated endocytosis might also participate in the transcytosis process.

The Use of a Microfluidic Device to Encapsulate a Poorly Water-Soluble Drug CoQ(10) in Lipid Nanoparticles and an Attempt to Regulate Intracellular Trafficking to Reach Mitochondria
Hibino Mitsue, Yamada Yuma, Fujishita Naoki, Sato Yusuke, Maeki Masatoshi, Tokeshi Manabu, Harashima Hideyoshi
JOURNAL OF PHARMACEUTICAL SCIENCES, 108, 8, 2668, 2676, Aug. 2019, [Peer-reviewed], [Last author, Corresponding author]
Scientific journal

A mitochondrial delivery system using liposome-based nanocarriers that target myoblast cells.
Katayama T, Kinugawa S, Takada S, Furihata T, Fukushima A, Yokota T, Anzai T, Hibino M, Harashima H, Yamada Y
Mitochondrion, 49, 66, 72, Jul. 2019, [Peer-reviewed]

Enhanced autophagy induction via the mitochondrial delivery of methylated beta-cyclodextrin-threaded polyrotaxanes using a MITO-Porter
Yuma Yamada, Shinnosuke Daikuhara, Atsushi Tamura, Kei Nishida, Nobuhiko Yui, Hideyoshi Harashima
CHEMICAL COMMUNICATIONS, 55, 50, 7203, 7206, Jun. 2019, [Peer-reviewed], [Last author]
English, Scientific journal

Lung-Endothelium-Targeted Nanoparticles Based on a pH-Sensitive Lipid and the GALA Peptide Enable Robust Gene Silencing and the Regression of Metastatic Lung Cancer
Mahmoud M. Abd Elwakil, Ikramy A. Khalil, Yaser H. A. Elewa, Kenji Kusumoto, Yusuke Sato, Nour Shobaki, Yasuhiro Kon, Hideyoshi Harashima
Advanced Functional Materials, 29, 18, Wiley-VCH Verlag, 02 May 2019
English, Scientific journal

MITO-Porterを用いた治療用mRNA送達によるミトコンドリア遺伝子治療戦略の検証　　　　　　　　　　　　　　　
山田 勇磨, 宗宮 加奈, 原島 秀吉
日本薬剤学会年会講演要旨集, 34年会, 179, 179, (公社)日本薬剤学会, May 2019
Japanese

Understanding structure-activity relationships of pH-sensitive cationic lipids facilitates the rational identification of promising lipid nanoparticles for delivering siRNAs in vivo
Sato Yusuke, Hashiba Kazuki, Sasaki Kosuke, Maeki Masatoshi, Tokeshi Manabu, Harashima Hideyoshi
JOURNAL OF CONTROLLED RELEASE, 295, 140, 152, Feb. 2019, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Development of a three-dimensional micromixer device for production of various lipid-based nucleic acid nanocarriers
Niko Kimura, Masatoshi Maeki, Yusuke Sato, Kosuke Sasaki, Akihiko Ishida, Hirofumi Tani, Hideyoshi Harashima, Manabu Tokeshi
23rd International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2019, 368, 369, 2019
International conference proceedings

Two modes of toxicity of lipid nanoparticles containing a pH-sensitive cationic lipid on human A375 and A375-SM melanoma cell lines　　　　　　　　　　　　　　　
Ahmed Y AlBaloul, Yusuke Sato, Nako Maishi, Kyoko Hida, Hideyoshi Harashima
BPB Reports, 2, 48, 55, 2019, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Critical parameters dictating efficiency of membrane-mediated drug transfer using nanoparticles.
Abbasi S, Kajimoto K, Harashima H
International journal of pharmaceutics, 553, 1-2, 398, 407, Dec. 2018, [Peer-reviewed], [Last author, Corresponding author]

Effective Therapy Using a Liposomal siRNA that Targets the Tumor Vasculature in a Model Murine Breast Cancer with Lung Metastasis.
Sakurai Y, Hada T, Kato A, Hagino Y, Mizumura W, Harashima H
Molecular therapy oncolytics, 11, 102, 108, Dec. 2018, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Although metastatic cancer is a major cause of death for cancer patients, no efficacious treatment for metastasis is available. We previously showed that the growth of a primary tumor could be inhibited by the administration of an anti-angiogenic small interfering RNA (siRNA) that is encapsulated in an RGD peptide-modified lipid nanoparticle (RGD-LNP). We herein report on the delivery of siRNA by an RGD-LNP to the vasculature is also effective for treating metastatic tumors. We compared the RGD-LNP with the polyethylene glycol (PEG)ylated LNP (PEG-LNP) in terms of accumulation in a lung-metastasized model. Despite malformed structure of vasculature in the metastasized lung, the accumulation of the PEG-LNP in the metastasized lung was lower than that for the RGD-LNP, which suggests that the delivery strategy based on vascular permeability is not completely effective for targeting metastasis tumors. The systemic injection of the RGD-LNP induced a significant silencing in the metastasized vasculature, but not in the normal lung. In addition, the continuous injection of the RGD-LNP encapsulating siRNA against a delta-like ligand 4 (DLL4) drastically prolonged the overall survival of metastasized model mice. Accordingly, our current findings suggest that vasculature targeting would be more effective than enhanced permeability and retention effect-based therapy for the treatment of metastatic cancer.

The delivery of mRNA to colon inflammatory lesions by lipid-nano-particles containing environmentally-sensitive lipid-like materials with oleic acid scaffolds.
Tanaka H, Watanabe A, Konishi M, Nakai Y, Yoshioka H, Ohkawara T, Takeda H, Harashima H, Akita H
Heliyon, 4, 12, e00959, Dec. 2018, [Peer-reviewed], [International Magazine]
English, Scientific journal, An mRNA gene therapy represents a potentially promising therapeutic for curing inflammatory diseases. The transient nature of the gene expression of mRNA would be expected to be beneficial for avoiding undesired side effects. Since the mRNA is a vulnerable molecule, a development of a carrier that can deliver the mRNA to the cytoplasm has a high priority. We report herein on the development of a system for delivering mRNA to the inflammatory lesion in a dextran sulfate sodium (DSS)-induced colitis model. We modulated molecular structures of an ionizable lipid, an SS-cleavable and pH-activated lipid-like material (ssPalm). Among the fatty acids investigated, oleic acid scaffolds (ssPalmO) appeared to be more biocompatible than either myristic acid or linoleic acid scaffolds with the colitis model. The structural modification of the hydrophilic head groups from linear tertiary amines to piperazine rings (ssPalmO-Paz4-C2) resulted in a more than 10-fold higher increasing in the transgene activity in inflammatory colon. The most notable observation is that the transgene activity in the inflammatory colon is significantly higher than that in liver, the major clearance organ of lipid nanoparticles. Collectively, the ssPalmO-Paz4-C2 represents a promising material for the delivery of an mRNA to inflammatory lesions.

ミトコンドリア強化幹細胞(MITO Cell)を用いた細胞移植療法の検証　　　　　　　　　　　　　　　
山田 勇磨, 阿部 二郎, 佐々木 大輔, 武田 充人, 原島 秀吉
日本バイオマテリアル学会大会予稿集, 40回, 180, 180, 日本バイオマテリアル学会, Nov. 2018
Japanese

DNA-loaded nano-adjuvant formed with a vitamin E-scaffold intracellular environmentally-responsive lipid-like material for cancer immunotherapy.
Kawai M, Nakamura T, Miura N, Maeta M, Tanaka H, Ueda K, Higashi K, Moribe K, Tange K, Nakai Y, Yoshioka H, Harashima H, Akita H
Nanomedicine : nanotechnology, biology, and medicine, 14, 8, 2587, 2597, Nov. 2018, [Peer-reviewed], [International Magazine]
English, Scientific journal, Cytoplasmic DNA triggers cellular immunity via activating the stimulator of interferon genes pathway. Since DNA is degradable and membrane impermeable, delivery system would permit cytoplasmic delivery by destabilizing the endosomal membrane for the use as an adjuvant. Herein, we report on the development of a plasmid DNA (pDNA)-encapsulating lipid nanoparticle (LNP). The structural components include an SS-cleavable and pH-activated lipid-like material that mounts vitamin E as a hydrophobic scaffold, and dual sensing motifs that are responsive to the intracellular environment (ssPalmE). The pDNA-encapsulating LNP (ssPalmE-LNP) induced a high interferon-β production in Raw 264.7 cells. The subcutaneous injection of ssPalmE-LNP strongly enhanced antigen-specific cytotoxic T cell activity. The ssPalmE-LNP treatment efficiently induced antitumor effects against E.G7-OVA tumor and B16-F10 melanoma metastasis. Furthermore, when combined with an anti-programmed death 1 antibody, an extensive therapeutic antitumor effect was observed. Therefore, the ssPalmE-LNP is a promising carrier of adjuvants for cancer immunotherapy.

Optimization of siRNA delivery to target sites: issues and future directions.
Khalil IA, Yamada Y, Harashima H
Expert opinion on drug delivery, 15, 11, 1053, 1065, Nov. 2018, [Peer-reviewed], [Invited], [Last author, Corresponding author]

Application of BCG-CWS as a Systemic Adjuvant by Using Nanoparticulation Technology.
Masuda H, Nakamura T, Noma Y, Harashima H
Molecular pharmaceutics, 15, 12, 5762, 5771, Nov. 2018, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, The intravesical instillation of live Bacillus Calmette-Guerin (BCG) for treating bladder cancer is a powerful cancer immunotherapy. The BCG cell wall skeleton (BCG-CWS) is the main component of the adjuvant, leading to the induction of antitumor immunity. However, the use of live BCG and BCG-CWS is currently limited to local administration because of the infectiousness of live BCG and the insolubility of BCG-CWS. We previously developed a water-dispersible nanoparticle (NP) formulation of BCG-CWS (CWS-NP), which could be used to apply BCG components for use as a systemically injected adjuvant for the treatment of cancers other than bladder cancer. In the present study, we examined the possible use of CWS-NP for cancer immunotherapy, when intravenously administered. The CWS-NP was a highly uniform dispersion and showed no aggregation in serum. The intravenously injected CWS-NP accumulated in the spleen and was efficiently taken up by dendritic cells, leading to their maturation. The coadministration of CWS-NP and ovalbumin (OVA) loaded NP resulted in the generation of OVA-specific cytotoxic T cells and inhibited the growth of E.G7-OVA tumors. These results represent the first findings related to the use of systemically injected CWS-NP as an adjuvant for cancer immunotherapy.

Neutralization of negative charges of siRNA results in improved safety and efficient gene silencing activity of lipid nanoparticles loaded with high levels of siRNA
Yusuke Sato, Hideki Matsui, Risa Sato, Hideyoshi Harashima
Journal of Controlled Release, 284, 179, 187, Elsevier B.V., 28 Aug. 2018, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

The use of cardiac progenitor cells for transplantation in congenital heart disease and an innovative strategy for activating mitochondrial function in such cells.
Abe J, Yamada Y, Harashima H
Journal of thoracic disease, 10, Suppl 18, S2119, S2121, Jul. 2018, [Peer-reviewed], [Last author, Corresponding author]

A hepatic pDNA delivery system based on an intracellular environment sensitive vitamin E-scaffold lipid-like material with the aid of an anti-inflammatory drug
Ryohei Togashi, Hiroki Tanaka, Sakiko Nakamura, Hideo Yokota, Kota Tange, Yuta Nakai, Hiroki Yoshioka, Hideyoshi Harashima, Hidetaka Akita
Journal of Controlled Release, 279, 262, 270, Elsevier B.V., 10 Jun. 2018, [Peer-reviewed]
English, Scientific journal

Reducing the Cytotoxicity of Lipid Nanoparticles Associated with a Fusogenic Cationic Lipid in a Natural Killer Cell Line by Introducing a Polycation-Based siRNA Core
Takashi Nakamura, Koharu Yamada, Yuki Fujiwara, Yusuke Sato, Hideyoshi Harashima
Molecular Pharmaceutics, 15, 6, 2142, 2150, American Chemical Society, 04 Jun. 2018, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Development of the iLiNP Device: Fine Tuning the Lipid Nanoparticle Size within 10 nm for Drug Delivery
Niko Kimura, Masatoshi Maeki, Yusuke Sato, Yusuke Note, Akihiko Ishida, Hirofumi Tani, Hideyoshi Harashima, Manabu Tokeshi
ACS Omega, 3, 5, 5044, 5051, American Chemical Society, 09 May 2018, [Peer-reviewed]
English, Scientific journal

In Vivo Introduction of mRNA Encapsulated in Lipid Nanoparticles to Brain Neuronal Cells and Astrocytes via Intracerebroventricular Administration.
Hiroki Tanaka, Taichi Nakatani, Tomomi Furihata, Kota Tange, Yuta Nakai, Hiroki Yoshioka, Hideyoshi Harashima, Hidetaka Akita
Molecular pharmaceutics, 15, 5, 2060, 2067, 07 May 2018, [Peer-reviewed], [International Magazine]
English, Scientific journal, Gene therapy is a promising strategy for curing certain types of brain diseases. Supplementation of therapeutic proteins such as aromatic amino acid decarboxylase (AADC) or nerve growth factor (NGF) have been reported to be successful examples of such treatments. However, there are safety concerns because these systems are based on virus-based gene vectors. A safe and efficient artificial carrier is thus urgently needed as an alternative. In this study, an mRNA based artificial gene carrier was introduced into the mouse brain via intracerebroventricular administration. As a carrier, a lipid nanoparticle (LNP) composed of environmentally sensitive lipid-like materials called an SS-cleavable proton-activated lipid-like material is used. The apolipoprotein E mediated cellular uptake of the lipid nanoparticles is one of the key features for its superior and homogeneous transfection activity compared to commercially available transfection reagents in both in vitro and in vivo situations. Immunostaining of brain specimens suggested that exogenous proteins can be introduced into neuronal cells as well as astrocytes using the mRNA-based gene carrier. This cannot be achieved using DNA-based artificial gene carriers. The findings suggest that a combination of an mRNA and a lipid based delivery system have great promise as a platform for the treatment of brain disorders.

微小化脂質ナノ粒子によるアジュバントのリンパ節送達　　　　　　　　　　　　　　　
中村 孝司, 河合 美典, 佐藤 悠介, 真栄城 正寿, 渡慶次 学, 原島 秀吉
日本薬剤学会年会講演要旨集, 33年会, 193, 193, (公社)日本薬剤学会, May 2018
Japanese

Synergism between a cell penetrating peptide and a pH-sensitive cationic lipid in efficient gene delivery based on double-coated nanoparticles
Ikramy A. Khalil, Seigo Kimura, Yusuke Sato, Hideyoshi Harashima
Journal of Controlled Release, 275, 107, 116, Elsevier B.V., 10 Apr. 2018, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Mitochondrial transgene expression via an artificial mitochondrial DNA vector in cells from a patient with a mitochondrial disease
Takuya Ishikawa, Kana Somiya, Reina Munechika, Hideyoshi Harashima, Yuma Yamada
Journal of Controlled Release, 274, 109, 117, Elsevier B.V., 28 Mar. 2018, [Peer-reviewed]
English, Scientific journal

Targeting the mitochondrial genome through a nanocarrier and the regulation of mitochondrial gene expression
Yuma Yamada, Hideyoshi Harashima
Mitochondrial Biology and Experimental Therapeutics, 491, 498, Springer International Publishing, 21 Mar. 2018, [Peer-reviewed], [Last author]
English, In book

Advances in microfluidics for lipid nanoparticles and extracellular vesicles and applications in drug delivery systems
Masatoshi Maeki, Niko Kimura, Yusuke Sato, Hideyoshi Harashima, Manabu Tokeshi
Advanced Drug Delivery Reviews, 128, 84, 100, Elsevier B.V., 15 Mar. 2018, [Peer-reviewed]
English

がん組織における浸潤性向上を目指した極小脂質ナノ粒子の開発　　　　　　　　　　　　　　　
岡部 奈々, 佐藤 悠介, 真栄城 正寿, 渡慶次 学, 原島 秀吉
日本薬学会年会要旨集, 138年会, 4, 78, 78, (公社)日本薬学会, Mar. 2018
Japanese

An efficient PEGylated gene delivery system with improved targeting: Synergism between octaarginine and a fusogenic peptide
Ikramy A. Khalil, Hideyoshi Harashima
International Journal of Pharmaceutics, 538, 1-2, 179, 187, Elsevier B.V., 01 Mar. 2018, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

In Vivo Transgene Expression in the Pancreas by the Intraductal Injection of Naked Plasmid DNA
Yuma Yamada, Mai Tabata, Jiro Abe, Masatoshi Nomura, Hideyoshi Harashima
Journal of Pharmaceutical Sciences, 107, 2, 647, 653, Elsevier B.V., 01 Feb. 2018, [Peer-reviewed], [Last author]
English, Scientific journal

Failure of active targeting by a cholesterol-anchored ligand and improvement by altering the lipid composition to prevent ligand desorption
Shoshiro Yamamoto, Yu Sakurai, Hideyoshi Harashima
International Journal of Pharmaceutics, 536, 1, 42, 49, Elsevier B.V., 30 Jan. 2018, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Mixing lipids to manipulate the ionization status of lipid nanoparticles for specific tissue targeting
Nour Shobaki, Yusuke Sato, Hideyoshi Harashima
International Journal of Nanomedicine, 13, 8395, 8410, Dove Medical Press Ltd., 2018
English, Scientific journal

A post-treatment methodology for precise size control of lipid nanoparticles by stepwise and rapid ethanol dilution
Niko Kimura, Masatoshi Maeki, Nana Okabe, Yusuke Sato, Akihiko Ishida, Hirofumi Tani, Hideyoshi Harashima, Manabu Tokeshi
22nd International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2018, 3, 1404, 1405, 2018
International conference proceedings

EPR effect and development of new strategy for nanoparticle delivery via remodeling tumor microenvironment based on tumor vasculature targeting
Yu Sakurai, Hideyoshi Harashima
Drug Delivery System, 33, 2, 98, 104, Japan Society of Drug Delivery System, 2018, [Peer-reviewed], [Invited], [Last author, Corresponding author]
Japanese, Scientific journal

Liposome Microencapsulation for the Surface Modification and Improved Entrapment of Cytochrome c for Targeted Delivery
Kazuaki Kajimoto, Tatsuhito Katsumi, Takashi Nakamura, Masatoshi Kataoka, Hideyoshi Harashima
JAOCS, Journal of the American Oil Chemists' Society, 95, 1, 101, 109, 01 Jan. 2018, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Cardiac progenitor cells activated by mitochondrial delivery of resveratrol enhance the survival of a doxorubicin-induced cardiomyopathy mouse model via the mitochondrial activation of a damaged myocardium
Jiro Abe, Yuma Yamada, Atsuhito Takeda, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 269, 177, 188, Jan. 2018, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Nano-sized drug carriers: Extravasation, intratumoral distribution, and their modeling
Joseph W. Nichols, Yu Sakurai, Hideyoshi Harashima, You Han Bae
JOURNAL OF CONTROLLED RELEASE, 267, 31, 46, Dec. 2017, [Peer-reviewed], [Invited]
English, Scientific journal

Preparation of envelope-type lipid nanoparticles containing gold nanorods for photothermal cancer therapy
West Kristian D. Paraiso, Hiroki Tanaka, Yusuke Sato, Daiki Shirane, Noriyuki Suzuki, Yasumitsu Ogra, Kota Tange, Yuta Nakai, Hiroki Yoshioka, Hideyoshi Harashima, Hidetaka Akita
COLLOIDS AND SURFACES B-BIOINTERFACES, 160, 715, 723, Dec. 2017, [Peer-reviewed]
English, Scientific journal

Optical control of mitochondrial reductive reactions in living cells using an electron donor-acceptor linked molecule
Yuta Takano, Reina Munechika, Vasudevanpillai Biju, Hideyoshi Harashima, Hiroshi Imahori, Yuma Yamada
NANOSCALE, 9, 47, 18690, 18698, Dec. 2017, [Peer-reviewed]
English, Scientific journal

Packaging of the Coenzyme Q(10) into a Liposome for Mitochondrial Delivery and the Intracellular Observation in Patient Derived Mitochondrial Disease Cells
Yuma Yamada, Laila Burger, Eriko Kawamura, Hideyoshi Harashima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 40, 12, 2183, 2190, Dec. 2017, [Peer-reviewed]
English, Scientific journal

Highly specific delivery of siRNA to hepatocytes circumvents endothelial cell-mediated lipid nanoparticle-associated toxicity leading to the safe and efficacious decrease in the hepatitis B virus
Yusuke Sato, Hideki Matsui, Naoki Yamamoto, Risa Sato, Tsubasa Munakata, Michinori Kohara, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 266, 216, 225, Nov. 2017, [Peer-reviewed]
English, Scientific journal

Integration of nano drug-delivery system with cancer immunotherapy
Takashi Nakamura, Hideyoshi Harashima
Therapeutic Delivery, 8, 11, 987, 1000, Future Medicine Ltd., 01 Nov. 2017, [Peer-reviewed]
English

Understanding the formation mechanism of lipid nanoparticles in microfluidic devices with chaotic micromixers
Masatoshi Maeki, Yuka Fujishima, Yusuke Sato, Takao Yasui, Noritada Kaji, Akihiko Ishida, Hirofumi Tani, Yoshinobu Baba, Hideyoshi Harashima, Manabu Tokeshi
PLOS ONE, 12, 11, e0187962, Nov. 2017, [Peer-reviewed]
English, Scientific journal

Identification and Evaluation of the Minimum Unit of a KALA Peptide Required for Gene Delivery and Immune Activation
Naoya Miura, Kota Tange, Yuta Nakai, Hiroki Yoshioka, Hideyoshi Harashima, Hidetaka Akita
JOURNAL OF PHARMACEUTICAL SCIENCES, 106, 10, 3113, 3119, Oct. 2017, [Peer-reviewed]
English, Scientific journal

Efficient siRNA Delivery by Lipid Nanoparticles Modified with a Nonstandard Macrocyclic Peptide for EpCAM-Targeting
Yu Sakurai, Wataru Mizumura, Manami Murata, Tomoya Hada, Shoshiro Yamamoto, Kenichiro Ito, Kazuhiro Iwasaki, Takayuki Katoh, Yuki Goto, Asako Takagi, Michinori Kohara, Hiroaki Suga, Hideyoshi Harashima
MOLECULAR PHARMACEUTICS, 14, 10, 3290, 3298, Oct. 2017, [Peer-reviewed]
English, Scientific journal

Mitochondrial Delivery of Doxorubicin Using MITO-Porter Kills Drug-Resistant Renal Cancer Cells via Mitochondrial Toxicity
Yuma Yamada, Reina Munechika, Eriko Kawamura, Yu Sakurai, Yusuke Sato, Hideyoshi Harashima
JOURNAL OF PHARMACEUTICAL SCIENCES, 106, 9, 2428, 2437, Sep. 2017, [Peer-reviewed]
English, Scientific journal

PEGylation of the GALA Peptide Enhances the Lung-Targeting Activity of Nanocarriers That Contain Encapsulated siRNA
Sarochin Santiwarangkool, Hidekata Akita, Taichi Nakatani, Kenji Kusumoto, Hiroki Kimura, Masaru Suzuki, Masaharu Nishimura, Yusuke Sato, Hideyoshi Harashima
JOURNAL OF PHARMACEUTICAL SCIENCES, 106, 9, 2420, 2427, Sep. 2017, [Peer-reviewed]
English, Scientific journal

pH-labile PEGylation of siRNA-loaded lipid nanoparticle improves active targeting and gene silencing activity in hepatocytes
Kazuki Hashiba, Yusuke Sato, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 262, 239, 246, Sep. 2017, [Peer-reviewed]
English, Scientific journal

Validation of the use of an artificial mitochondrial reporter DNA vector containing a Cytomegalovirus promoter for mitochondrial transgene expression
Yuma Yamada, Takuya Ishikawa, Hideyoshi Harashima
BIOMATERIALS, 136, 56, 66, Aug. 2017, [Peer-reviewed]
English, Scientific journal

ドキソルビシン心筋症に対するミトコンドリアドラッグデリバリーシステムを用いた心筋前駆細胞移植の検討　　　　　　　　　　　　　　　
阿部 二郎, 山田 勇磨, 武田 充人, 原島 秀吉
日本小児循環器学会雑誌, 33, Suppl.1, s1, 175, (NPO)日本小児循環器学会, Jul. 2017
Japanese

Size-Dependency of the Surface Ligand Density of Liposomes Prepared by Post-insertion
Shang-Hsuan Lee, Yusuke Sato, Mamoru Hyodo, Hideyoshi Harashima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 40, 7, 1002, 1009, Jul. 2017, [Peer-reviewed]
English, Scientific journal

リンパ節内樹状細胞を標的とした極小ナノキャリアシステムの開発　　　　　　　　　　　　　　　
河合 美典, 中村 孝司, 佐藤 悠介, 真栄城 正寿, 原島 秀吉
日本DDS学会学術集会プログラム予稿集, 33回, 175, 175, 日本DDS学会, Jun. 2017
Japanese

オートファジー誘導能を有する超分子構造ポリマー搭載ミトコンドリアDDSの構築　　　　　　　　　　　　　　　
大工原 伸之輔, 山田 勇磨, 田村 篤志, 西田 慶, 由井 伸彦, 原島 秀吉
日本DDS学会学術集会プログラム予稿集, 33回, 173, 173, 日本DDS学会, Jun. 2017, [Peer-reviewed]
Japanese

Modality of tumor endothelial VEGFR2 silencing-mediated improvement in intratumoral distribution of lipid nanoparticles
Shoshiro Yamamoto, Akari Kato, Yu Sakurai, Tomoya Hada, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 251, 1, 10, Apr. 2017, [Peer-reviewed]
English, Scientific journal

Modifying Antigen-Encapsulating Liposomes with KALA Facilitates MHC Class I Antigen Presentation and Enhances Anti-tumor Effects
Naoya Miura, Hidetaka Akita, Naho Tateshita, Takashi Nakamura, Hideyoshi Harashima
MOLECULAR THERAPY, 25, 4, 1003, 1013, Apr. 2017, [Peer-reviewed]
English, Scientific journal

Enhanced transgene expression by plasmid-specific recruitment of histone acetyltransferase
Mika Nishihara, Genki N. Kanda, Tetsuya Suzuki, Shin'ichiro Yamakado, Hideyoshi Harashima, Hiroyuki Kamiya
JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 123, 3, 277, 280, Mar. 2017, [Peer-reviewed]
English, Scientific journal

Temperature and pH sensitivity of a stabilized self-nanoemulsion formed using an ionizable lipid-like material via an oil-to-surfactant transition
Hiroki Tanaka, Sho Oasa, Masataka Kinjo, Kota Tange, Yuta Nakai, Hideyoshi Harashima, Hidetaka Akita
COLLOIDS AND SURFACES B-BIOINTERFACES, 151, 95, 101, Mar. 2017, [Peer-reviewed]
English, Scientific journal

Modifying Cationic Liposomes with Cholesteryl-PEG Prevents Their Aggregation in Human Urine and Enhances Cellular Uptake by Bladder Cancer Cells
Takashi Nakamura, Yosuke Noma, Yu Sakurai, Hideyoshi Harashima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 40, 2, 234, 237, Feb. 2017, [Peer-reviewed]
English, Scientific journal

MITO-Porter, multifunctional envelope-type nano device for mitochondrial delivery toward innovative nano medicine
Yuma Yamada, Hideyoshi Harashima
Journal of the Society of Powder Technology, Japan, 54, 3, 158, 166, Society of Powder Technology, 2017, [Peer-reviewed]
Japanese, Scientific journal

Innovative Technologies in Nanomedicines: From Passive Targeting to Active Targeting/From Controlled Pharmacokinetics to Controlled Intracellular Pharmacokinetics
Yusuke Sato, Yu Sakurai, Kazuaki Kajimoto, Takashi Nakamura, Yuma Yamada, Hidetaka Akita, Hideyoshi Harashima
MACROMOLECULAR BIOSCIENCE, 17, 1, Jan. 2017, [Peer-reviewed]
English, Scientific journal

MITO-porter for mitochondrial delivery and mitochondrial functional analysis
Yuma Yamada, Hideyoshi Harashima
Handbook of Experimental Pharmacology, 240, 457, 472, Springer New York LLC, 2017, [Peer-reviewed]
English, In book

Drug Delivery Research for the Future: Expanding the Nano Horizons and Beyond
Samir Mitragotri, Twan Lammers, You Han Bae, Steven Schwendeman, Stefaan De Smedt, Jean-Christophe Leroux, Dan Peer, Ick Chan Kwon, Hideyoshi Harashima, Akihiko Kikuchi, Yu-Kyoung Oh, Vladmir Torchilin, Wim Hennink, Justin Hanes, Kinam Park
JOURNAL OF CONTROLLED RELEASE, 246, 183, 184, Jan. 2017, [Peer-reviewed]
English

Scalable preparation of poly(ethyleneglycol)-grafted siRNA-loaded lipid nanoparticles using a commercially available fluidic device and tangential flow filtration
Yu Sakurai, Tomoya Hada, Hideyoshi Harashima
JOURNAL OF BIOMATERIALS SCIENCE-POLYMER EDITION, 28, 10-12, 1086, 1096, 2017, [Peer-reviewed]
English, Scientific journal

Remodeling of the extracellular matrix by endothelial cell-targeting siRNA improves the EPR-based delivery of 100 nm particles
Yu Sakurai, Tomoya Hada, Shoshiro Yamamoto, Akari Kato, Wataru Mizumura, Hideyoshi Harashima
Molecular Therapy, 24, 12, 2090, 2099, Nature Publishing Group, 01 Dec. 2016, [Peer-reviewed]
English, Scientific journal

Development of a multifunctional envelope-type nano device and its application to nanomedicine
Yusuke Sato, Takashi Nakamura, Yuma Yamada, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 244, Pt B, 194, 204, Dec. 2016, [Peer-reviewed]
English, Scientific journal

Remodeling of the Extracellular Matrix by Endothelial Cell-Targeting siRNA Improves the EPR-Based Delivery of 100 nm Particles
Yu Sakurai, Tomoya Hada, Shoshiro Yamamoto, Akari Kato, Wataru Mizumura, Hideyoshi Harashima
MOLECULAR THERAPY, 24, 12, 2090, 2099, Dec. 2016, [Peer-reviewed]
English, Scientific journal

Topology of Surface Ligands on Liposomes: Characterization Based on the Terms, Incorporation Ratio, Surface Anchor Density, and Reaction Yield
Shang-Hsuan Lee, Yusuke Sato, Mamoru Hyodo, Hideyoshi Harashima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 39, 12, 1983, 1994, Dec. 2016, [Peer-reviewed]
English, Scientific journal

Small-sized, stable lipid nanoparticle for the efficient delivery of siRNA to human immune cell lines
Takashi Nakamura, Moeka Kuroi, Yuki Fujiwara, Shota Warashina, Yusuke Sato, Hideyoshi Harashima
SCIENTIFIC REPORTS, 6, 37849, Nov. 2016, [Peer-reviewed]
English, Scientific journal

Identification of a Nucleoporin358-Specific RNA Aptamer for Use as a Nucleus-Targeting Liposomal Delivery System
Garima Shrivastava, Mamoru Hyodo, Shige H. Yoshimura, Hidetaka Akita, Hideyoshi Harashima
NUCLEIC ACID THERAPEUTICS, 26, 5, 286, 298, Oct. 2016, [Peer-reviewed]
English, Scientific journal

Liver-Specific Silencing of Lipin1 Reduces Fat Mass as Well as Hepatic Triglyceride Biosynthesis in Mice
Kazuaki Kajimoto, Erina Suemitsu, Yusuke Sato, Yu Sakurai, Hideyoshi Harashima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 39, 10, 1653, 1661, Oct. 2016, [Peer-reviewed]
English, Scientific journal

ミトコンドリア薬物送達システムを用いた心筋炎に対する新たな治療戦略　　　　　　　　　　　　　　　
阿部 二郎, 山田 勇磨, 武田 充人, 原島 秀吉
日本小児循環器学会雑誌, 32, Suppl.1, s1, 208, (NPO)日本小児循環器学会, Jul. 2016, [Peer-reviewed]
Japanese

Effect of particle size on their accumulation in an inflammatory lesion in a dextran sulfate sodium (DSS)-induced colitis model
Ayaka Watanabe, Hiroki Tanaka, Yu Sakurai, Kota Tange, Yuta Nakai, Tatsuya Ohkawara, Hiroshi Takeda, Hideyoshi Harashima, Hidetaka Akita
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 509, 1-2, 118, 122, Jul. 2016, [Peer-reviewed]
English, Scientific journal

Anti-tumor effect via passive anti-angiogenesis of PEGylated liposomes encapsulating doxorubicin in drug resistant tumors
Golam Kibria, Hiroto Hatakeyama, Yusuke Sato, Hideyoshi Harashima
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 509, 1-2, 178, 187, Jul. 2016, [Peer-reviewed]
English, Scientific journal

Elimination of the biphasic pharmacodynamics of 15d-PGJ2 by controlling its release from a nanoemulsion
Saed Abbasi, Kazuaki Kajimoto, Hideyoshi Harashima
International Journal of Nanomedicine, 11, 2685, 2694, Dove Medical Press Ltd., 08 Jun. 2016, [Peer-reviewed]
English, Scientific journal

プラスミドDNA結合ヒストンの特異的アセチル化による外来遺伝子発現上昇　　　　　　　　　　　　　　　
紙谷 浩之, 西原 実香, 神田 元紀, 鈴木 哲矢, 山門 振一郎, 原島 秀吉
日本DDS学会学術集会プログラム予稿集, 32回, 153, 153, 日本DDS学会, Jun. 2016
Japanese

Protecting liver sinusoidal endothelial cells suppresses apoptosis in acute liver damage
Tomohito Tanoi, Takafumi Tamura, Naoki Sano, Ken Nakayama, Kiyoshi Fukunaga, Yun-Wen Zheng, Afsana Akhter, Yu Sakurai, Yasuhiro Hayashi, Hideyoshi Harashima, Nobuhiro Ohkohchi
HEPATOLOGY RESEARCH, 46, 7, 697, 706, Jun. 2016, [Peer-reviewed]
English, Scientific journal

In vivo selection of active deoxyribonucleoside kinase by a mutagenic nucleoside analog
Hiroyuki Kamiya, Mana Ito, Kosuke Nishi, Hideyoshi Harashima
JOURNAL OF BIOTECHNOLOGY, 228, 52, 57, Jun. 2016, [Peer-reviewed]
English, Scientific journal

Elucidation of the physicochemical properties and potency of siRNA-loaded small-sized lipid nanoparticles for siRNA delivery
Yusuke Sato, Yusuke Note, Masatoshi Maeki, Noritada Kaji, Yoshinobu Baba, Manabu Tokeshi, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 229, 48, 57, May 2016, [Peer-reviewed]
English, Scientific journal

A Dual-Ligand Liposomal System Composed of a Cell-Penetrating Peptide and a Mitochondrial RNA Aptamer Synergistically Facilitates Cellular Uptake and Mitochondrial Targeting
Yuma Yamada, Ryo Furukawa, Hideyoshi Harashima
JOURNAL OF PHARMACEUTICAL SCIENCES, 105, 5, 1705, 1713, May 2016, [Peer-reviewed]
English, Scientific journal

Heterogeneity of tumor endothelial cells and drug delivery
Kyoko Hida, Nako Maishi, Yu Sakurai, Yasuhiro Hida, Hideyoshi Harashima
ADVANCED DRUG DELIVERY REVIEWS, 99, Pt B, 140, 147, Apr. 2016, [Peer-reviewed]
English

Relationship Between the Physicochemical Properties of Lipid Nanoparticles and the Quality of siRNA Delivery to Liver Cells
Yusuke Sato, Hiroto Hatakeyama, Mamoru Hyodo, Hideyoshi Harashima
MOLECULAR THERAPY, 24, 4, 788, 795, Apr. 2016, [Peer-reviewed]
English, Scientific journal

Tri-membrane nanoparticles produced by combining liposome fusion and a novel patchwork of bicelles to overcome endosomal and nuclear membrane barriers to cargo delivery
Asako Yamada, Asako Mitsueda, Mahadi Hasan, Miho Ueda, Susumu Hama, Shota Warashina, Takashi Nakamura, Hideyoshi Harashima, Kentaro Kogure
Biomaterials Science, 4, 3, 439, 447, Royal Society of Chemistry, 01 Mar. 2016, [Peer-reviewed]
English, Scientific journal

Transcytosis-Targeting Peptide: A Conductor of Liposomal Nanoparticles through the Endothelial Cell Barrier
Hidetaka Akita, Takahiro Fujiwara, Sarochin Santiwarangkool, Nazir Hossen, Kazuaki Kajimoto, Ayman El-Sayed, Yasuhiko Tabata, Hideyoshi Harashima
SMALL, 12, 9, 1212, 1221, Mar. 2016, [Peer-reviewed]
English, Scientific journal

Novel pH-sensitive multifunctional envelope-type nanodevice for siRNA-based treatments for chronic HBV infection
Naoki Yamamoto, Yusuke Sato, Tsubasa Munakata, Masakazu Kakuni, Chise Tateno, Takahiro Sanada, Yuichi Hirata, Shuko Murakami, Yasuhito Tanaka, Kazuaki Chayama, Hiroto Hatakeyama, Mamoru Hyodo, Hideyoshi Harashima, Michinori Kohara
JOURNAL OF HEPATOLOGY, 64, 3, 547, 555, Mar. 2016, [Peer-reviewed]
English, Scientific journal

FoxO1 Plays an Important Role in Regulating beta-Cell Compensation for Insulin Resistance in Male Mice
Ting Zhang, Dae Hyun Kim, Xiangwei Xiao, Sojin Lee, Zhenwei Gong, Radhika Muzumdar, Virtu Calabuig-Navarro, Jun Yamauchi, Hideyoshi Harashima, Rennian Wang, Rita Bottino, Juan Carlos Alvarez-Perez, Adolfo Garcia-Ocana, George Gittes, H. Henry Dong
ENDOCRINOLOGY, 157, 3, 1055, 1070, Mar. 2016, [Peer-reviewed]
English, Scientific journal

A lipid nanoparticle for the efficient delivery of siRNA to dendritic cells
Shota Warashina, Takashi Nakamura, Yusuke Sato, Yuki Fujiwara, Mamoru Hyodo, Hiroto Hatakeyama, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 225, 183, 191, Mar. 2016, [Peer-reviewed]
English, Scientific journal

Validation of a Strategy for Cancer Therapy: Delivering Aminoglycoside Drugs to Mitochondria in HeLa Cells
Jiro Abe, Yuma Yamada, Hideyoshi Harashima
JOURNAL OF PHARMACEUTICAL SCIENCES, 105, 2, 734, 740, Feb. 2016, [Peer-reviewed]
English, Scientific journal

PEG dilemma- nucleic acids delivery to cancers by controlling biodistribution and intracellular trafficking
Hiroto Hatakeyama, Hideyoshi Harashima
Drug Delivery System, 31, 4, 293, 299, Japan Society of Drug Delivery System, 2016, [Peer-reviewed]
Japanese, Scientific journal

Elimination of the biphasic pharmacodynamics of 15d-PGJ2 by controlling its release from a nanoemulsion
Saed Abbasi, Kazuaki Kajimoto, Hideyoshi Harashima
INTERNATIONAL JOURNAL OF NANOMEDICINE, 11, 2685, 2694, 2016, [Peer-reviewed]
English, Scientific journal

Preparation of a Cyclic RGD: Modified Liposomal SiRNA Formulation for Use in Active Targeting to Tumor and Tumor Endothelial Cells.
Sakurai Y, Hada T, Harashima H
Methods in molecular biology (Clifton, N.J.), 1364, 63, 69, 2016, [Peer-reviewed]

A Multifunctional Envelope-Type Nano Device Containing a pH-Sensitive Cationic Lipid for Efficient Delivery of Short Interfering RNA to Hepatocytes In Vivo.
Sato Y, Harashima H, Kohara M
Methods in molecular biology (Clifton, N.J.), 1364, 71, 78, 2016, [Peer-reviewed]

Tri-membrane nanoparticles produced by combining liposome fusion and a novel patchwork of bicelles to overcome endosomal and nuclear membrane barriers to cargo delivery
Asako Yamada, Asako Mitsueda, Mahadi Hasan, Miho Ueda, Susumu Hama, Shota Warashina, Takashi Nakamura, Hideyoshi Harashima, Kentaro Kogure
BIOMATERIALS SCIENCE, 4, 3, 439, 447, 2016, [Peer-reviewed]
English, Scientific journal

In Vivo Inverse Correlation in the Activation of Natural Killer T Cells Through Dual-Signal Stimulation via a Combination of alpha-Galactosylceramide-Loaded Liposomes and Interleukin-12
Heba Abdelmegeed, Takashi Nakamura, Hideyoshi Harashima
JOURNAL OF PHARMACEUTICAL SCIENCES, 105, 1, 250, 256, Jan. 2016, [Peer-reviewed]
English, Scientific journal

Cellular environment-responsive nanomaterials for use in gene and siRNA delivery: molecular design for biomembrane destabilization and intracellular collapse
Hiroki Tanaka, Yusuke Sato, Hideyoshi Harashima, Hidetaka Akita
EXPERT OPINION ON DRUG DELIVERY, 13, 7, 1015, 1027, 2016, [Peer-reviewed]
English

Insertion and Deletion Mismatches Distant from the Target Position Improve Gene Correction with a Tailed Duplex
Hiroyuki Kamiya, Natsuki Nishigaki, Akihiro Ikeda, Seiya Yukawa, Yukiko Morita, Yoshimichi Nakatsu, Teruhisa Tsuzuki, Hideyoshi Harashima
NUCLEOSIDES NUCLEOTIDES & NUCLEIC ACIDS, 35, 7, 379, 388, 2016, [Peer-reviewed]
English, Scientific journal

Validation of Mitochondrial Gene Delivery in Liver and Skeletal Muscle via Hydrodynamic Injection Using an Artificial Mitochondrial Reporter DNA Vector
Yukari Yasuzaki, Yuma Yamada, Takuya Ishikawa, Hideyoshi Harashima
MOLECULAR PHARMACEUTICS, 12, 12, 4311, 4320, Dec. 2015, [Peer-reviewed]
English, Scientific journal

New drug delivery system for liver sinusoidal endothelial cells for ischemia-reperfusion injury
Naoki Sano, Takafumi Tamura, Naoyuki Toriyabe, Takeshi Nowatari, Ken Nakayama, Tomohito Tanoi, Soichiro Murata, Yu Sakurai, Mamoru Hyodo, Kiyoshi Fukunaga, Hideyoshi Harashima, Nobuhiro Ohkohchi
WORLD JOURNAL OF GASTROENTEROLOGY, 21, 45, 12778, 12786, Dec. 2015, [Peer-reviewed]
English, Scientific journal

Size-dependent specific targeting and efficient gene silencing in peritoneal macrophages using a pH-sensitive cationic liposomal siRNA carrier
Hideki Matsui, Yusuke Sato, Hiroto Hatakeyama, Hidetaka Akita, Hideyoshi Harashima
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 495, 1, 171, 178, Nov. 2015, [Peer-reviewed]
English, Scientific journal

The intracellular pharmacodynamics of siRNA is responsible for the low gene silencing activity of siRNA-loaded nanoparticles in dendritic cells
Takashi Nakamura, Yuki Fujiwara, Shota Warashina, Hideyoshi Harashima
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 494, 1, 271, 277, Oct. 2015, [Peer-reviewed]
English, Scientific journal

Liposomes loaded with a STING pathway ligand, cyclic di-GMP, enhance cancer immunotherapy against metastatic melanoma
Takashi Nakamura, Hiroko Miyabe, Mamoru Hyodo, Yusuke Sato, Yoshihiro Hayakawa, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 216, 149, 157, Oct. 2015, [Peer-reviewed]
English, Scientific journal

Molecular Tuning of a Vitamin E-Scaffold pH-Sensitive and Reductive Cleavable Lipid-like Material for Accelerated in Vivo Hepatic siRNA Delivery
Hidetaka Akita, Yuki Noguchi, Hiroto Hatakeyama, Yusuke Sato, Kota Tange, Yuta Nakai, Hideyoshi Harashima
ACS BIOMATERIALS SCIENCE & ENGINEERING, 1, 9, 834, 844, Sep. 2015, [Peer-reviewed]
English, Scientific journal

Mitochondrial delivery of Coenzyme Q(10) via systemic administration using a MITO-Porter prevents ischemia/reperfusion injury in the mouse liver
Yuma Yamada, Kohei Nakamura, Jiro Abe, Mamoru Hyodo, Sanae Haga, Michitaka Ozaki, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 213, 86, 95, Sep. 2015, [Peer-reviewed]
English, Scientific journal

An analysis of membrane fusion between mitochondrial double membranes and MITO-Porter, mitochondrial fusogenic vesicles
Yuma Yamada, Yutaka Fukuda, Hideyoshi Harashima
MITOCHONDRION, 24, 50, 55, Sep. 2015, [Peer-reviewed]
English, Scientific journal

Efficient and High-Speed Transduction of an Antibody into Living Cells Using a Multifunctional Nanocarrier System to Control Intracellular Trafficking
Yuma Yamada, Sandra Milena Vergara Perez, Mai Tabata, Jiro Abe, Yukari Yasuzaki, Hideyoshi Harashima
JOURNAL OF PHARMACEUTICAL SCIENCES, 104, 9, 2845, 2854, Sep. 2015, [Peer-reviewed]
English, Scientific journal

Efficient Packaging of Plasmid DNA Using a pH Sensitive Cationic Lipid for Delivery to Hepatocytes
Yu Sakurai, Takashi Matsuda, Tomoya Hada, Hideyoshi Harashima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 38, 8, 1185, 1191, Aug. 2015, [Peer-reviewed]
English, Scientific journal

Influence of Endosomal Escape and Degradation of alpha-Galactosylceramide Loaded Liposomes on CD1d Antigen Presentation
Takashi Nakamura, Moeka Kuroi, Hideyoshi Harashima
MOLECULAR PHARMACEUTICS, 12, 8, 2791, 2799, Aug. 2015, [Peer-reviewed]
English, Scientific journal

Mitochondrial delivery of antisense RNA by MITO-Porter results in mitochondrial RNA knockdown, and has a functional impact on mitochondria
Ryo Furukawa, Yuma Yamada, Eriko Kawamura, Hideyoshi Harashima
BIOMATERIALS, 57, 107, 115, Jul. 2015, [Peer-reviewed]
English, Scientific journal

Multifunctional Envelope-Type Nano Device: Evolution from Nonselective to Active Targeting System
Yasuhiro Hayashi, Hiroto Hatakeyama, Kazuaki Kajimoto, Mamoru Hyodo, Hidetaka Akita, Hideyoshi Harashima
BIOCONJUGATE CHEMISTRY, 26, 7, 1266, 1276, Jul. 2015, [Peer-reviewed]
English

General considerations regarding the in vitro and in vivo properties of block copolymer micelle products and their evaluation
Kumiko Sakai-Kato, Nobuhiro Nishiyama, Masato Kozaki, Takeshi Nakanishi, Yoshihiro Matsuda, Mai Hirano, Hiroyuki Hanada, Shigeru Hisada, Hiroshi Onodera, Hideyoshi Harashima, Yasuhiro Matsumura, Kazunori Kataoka, Yukihiro Goda, Haruhiro Okuda, Toru Kawanishi
JOURNAL OF CONTROLLED RELEASE, 210, 76, 83, Jul. 2015, [Peer-reviewed]
English

Effect of hydrophobic scaffold on the cellular uptake and gene transfection activities of DNA-encapsulating liposomal nanoparticles via intracerebroventricular administration
Hidetaka Akita, Taichi Nakatani, Kimiko Kuroki, Katsumi Maenaka, Kota Tange, Yuta Nakai, Hideyoshi Harashima
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 490, 1-2, 142, 145, Jul. 2015, [Peer-reviewed]
English, Scientific journal

Hyaluronic acid controls the uptake pathway and intracellular trafficking of an octaarginine-modified gene vector in CD44 positive- and CD44 negative-cells
Yuma Yamada, Masahiro Hashida, Hideyoshi Harashima
BIOMATERIALS, 52, 189, 198, Jun. 2015, [Peer-reviewed]
English, Scientific journal

Forkhead Box O6 (FoxO6) Depletion Attenuates Hepatic Gluconeogenesis and Protects against Fat-induced Glucose Disorder in Mice
Virtu Calabuig-Navarro, Jun Yamauchi, Sojin Lee, Ting Zhang, Yun-Zi Liu, Kelsey Sadlek, Gina M. Coudriet, Jon D. Piganelli, Chun-Lei Jiang, Rita Miller, Mark Lowe, Hideyoshi Harashima, H. Henry Dong
JOURNAL OF BIOLOGICAL CHEMISTRY, 290, 25, 15581, 15594, Jun. 2015, [Peer-reviewed]
English, Scientific journal

Effect of the Compaction and the Size of DNA on the Nuclear Transfer Efficiency after Microinjection in Synchronized Cells
Hidetaka Akita, Dai Kurihara, Marco Schmeer, Martin Schleef, Hideyoshi Harashima
PHARMACEUTICS, 7, 2, 64, 73, Jun. 2015, [Peer-reviewed]
English, Scientific journal

Action-at-a-Distance Mutagenesis Induced by Oxidized Guanine in Werner Syndrome Protein-Reduced Human Cells
Hiroyuki Kamiya, Daiki Yamazaki, Eri Nakamura, Tetsuaki Makino, Miwako Kobayashi, Ichiro Matsuoka, Hideyoshi Harashima
CHEMICAL RESEARCH IN TOXICOLOGY, 28, 4, 621, 628, Apr. 2015, [Peer-reviewed]
English, Scientific journal

A neutral lipid envelope-type nanoparticle composed of a pH-activated and vitamin E-scaffold lipid-like material as a platform for a gene carrier targeting renal cell carcinoma
Hidetaka Akita, Ryohei Ishiba, Ryohei Togashi, Kota Tange, Yuta Nakai, Hiroto Hatakeyama, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 200, 97, 105, Feb. 2015, [Peer-reviewed]
English, Scientific journal

A KALA-modified lipid nanoparticle containing CpG-free plasmid DNA as a potential DNA vaccine carrier for antigen presentation and as an immune-stimulative adjuvant
Naoya Miura, Sharif M. Shaheen, Hidetaka Akita, Takashi Nakamura, Hideyoshi Harashima
NUCLEIC ACIDS RESEARCH, 43, 3, 1317, 1331, Feb. 2015, [Peer-reviewed]
English, Scientific journal

Targeting the mitochondrial genome via a dual function MITO-porter: Evaluation of mtDNA levels and mitochondrial function
Yuma Yamada, Hideyoshi Harashima
Mitochondrial Medicine, 2, 123, 133, Springer New York, 29 Jan. 2015, [Peer-reviewed]
English, In book

Optimization of a siRNA carrier modified with a ph-sensitive cationic lipid and a cyclic RGD peptide for efficiently targeting tumor endothelial cells
Tomoya Hada, Yu Sakurai, Hideyoshi Harashima
Pharmaceutics, 7, 3, 320, 333, MDPI AG, 2015, [Peer-reviewed]
English, Scientific journal

A strategy for synthesis of lipid nanoparticles using microfluidic devices with a mixer structure
Masatoshi Maeki, Tatsuyoshi Saito, Yusuke Sato, Takao Yasui, Noritada Kaji, Akihiko Ishida, Hirofumi Tani, Yoshinobu Baba, Hideyoshi Harashima, Manabu Tokeshi
RSC ADVANCES, 5, 57, 46181, 46185, 2015, [Peer-reviewed]
English, Scientific journal

Advances in an active and passive targeting to tumor and adipose tissues
Yu Sakurai, Kazuaki Kajimoto, Hiroto Hatakeyama, Hideyoshi Harashima
EXPERT OPINION ON DRUG DELIVERY, 12, 1, 41, 52, Jan. 2015, [Peer-reviewed]
English

The RNA Sensor RIG-I Dually Functions as an Innate Sensor and Direct Antiviral Factor for Hepatitis B Virus
Seiichi Sato, Kai Li, Takeshi Kameyama, Takaya Hayashi, Yuji Ishida, Shuko Murakami, Tsunamasa Watanabe, Sayuki Iijima, Yu Sakurai, Koichi Watashi, Susumu Tsutsumi, Yusuke Sato, Hidetaka Akita, Takaji Wakita, Charles M. Rice, Hideyoshi Harashima, Michinori Kohara, Yasuhito Tanaka, Akinori Takaoka
IMMUNITY, 42, 1, 123, 132, Jan. 2015, [Peer-reviewed]
English, Scientific journal

Mechanism responsible for the antitumor effect of BCG-CWS using the LEEL method in a mouse bladder cancer model
Takashi Nakamura, Masafumi Fukiage, Yoshiteru Suzuki, Ikuya Yano, Jun Miyazaki, Hiroyuki Nishiyama, Hideyuki Akaza, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 196, 161, 167, Dec. 2014, [Peer-reviewed]
English, Scientific journal

Comparative Study of the Sensitivities of Cancer Cells to Doxorubicin, and Relationships between the Effect of the Drug-Efflux Pump P-gp
Golam Kibria, Hiroto Hatakeyama, Kosuke Akiyama, Kyoko Hida, Hideyoshi Harashima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 37, 12, 1926, 1935, Dec. 2014, [Peer-reviewed]
English, Scientific journal

"Programmed packaging" for gene delivery
M. Hyodo, Y. Sakurai, H. Akita, H. Harashima
JOURNAL OF CONTROLLED RELEASE, 193, 316, 323, Nov. 2014, [Peer-reviewed]
English, Scientific journal

Ligand density at the surface of a nanoparticle and different uptake mechanism: Two important factors for successful siRNA delivery to liver endothelial cells
Afsana Akhter, Yasuhiro Hayashi, Yu Sakurai, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 475, 1-2, 227, 237, Nov. 2014, [Peer-reviewed]
English, Scientific journal

The Screening of RNA Aptamers Specific for Carbonic Anhydrase I Using the Systematic Evolution of Ligands by an Exponential Enrichment Method (SELEX)
Garima Shrivastava, Mamoru Hyodo, Mst Naznin Ara, Hideyoshi Harashima
NUCLEOSIDES NUCLEOTIDES & NUCLEIC ACIDS, 33, 11, 697, 708, Nov. 2014, [Peer-reviewed]
English, Scientific journal

Construction of an Aptamer Modified Liposomal System Targeted to Tumor Endothelial Cells
Mst Naznin Ara, Takashi Matsuda, Mamoru Hyodo, Yu Sakurai, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 37, 11, 1742, 1749, Nov. 2014, [Peer-reviewed]
English, Scientific journal

Advantages of ethanol dilution method for preparing GALA-modified liposomal siRNA carriers on the in vivo gene knockdown efficiency in pulmonary endothelium
Kenji Kusumoto, Hidetaka Akita, Sarochin Santiwarangkool, Hideyoshi Harashima
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 473, 1-2, 144, 147, Oct. 2014, [Peer-reviewed]
English, Scientific journal

Multifunctional envelope-type nano device for controlled intracellular trafficking and selective targeting in vivo
Kazuaki Kajimoto, Yusuke Sato, Takashi Nakamura, Yuma Yamada, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 190, 593, 606, Sep. 2014, [Peer-reviewed]
English

Condensation of plasmid DNA enhances mitochondrial association in skeletal muscle following hydrodynamic limb vein injection
Yukari Yasuzaki, Yuma Yamada, Yutaka Fukuda, Hideyoshi Harashima
Pharmaceuticals, 7, 8, 881, 893, 21 Aug. 2014, [Peer-reviewed]
English, Scientific journal

Neutralized Nanoparticle Composed of SS-Cleavable and pH-Activated Lipid-Like Material as a Long-Lasting and Liver-Specific Gene Delivery System
Masami Ukawa, Hidetaka Akita, Yasuhiro Hayashi, Ryohei Ishiba, Kota Tange, Masaya Arai, Kazuhiro Kubo, Yuriko Higuchi, Kazunori Shimizu, Satoshi Konishi, Mitsuru Hashida, Hideyoshi Harashima
ADVANCED HEALTHCARE MATERIALS, 3, 8, 1222, 1229, Aug. 2014, [Peer-reviewed]
English, Scientific journal

Improvement of Doxorubicin Efficacy Using Liposornal Anti-Polo-like Kinase 1 siRNA in Human Renal Cell Carcinomas
Yu Sakurai, Hiroto Hatakeyama, Hidetaka Akita, Hideyoshi Harashima
MOLECULAR PHARMACEUTICS, 11, 8, 2713, 2719, Aug. 2014, [Peer-reviewed]
English, Scientific journal

Comparative assessments of crucial factors for a functional ligand-targeted nanocarrier
Md. Nazir Hossen, Kazuaki Kajimoto, Ryo Tatsumi, Mamoru Hyodo, Hideyoshi Harashima
JOURNAL OF DRUG TARGETING, 22, 7, 600, 609, Aug. 2014, [Peer-reviewed]
English, Scientific journal

Identification and expression of troponin T, a new marker on the surface of cultured tumor endothelial cells by aptamer ligand
Mst. Naznin Ara, Mamoru Hyodo, Noritaka Ohga, Kosuke Akiyama, Kyoko Hida, Yasuhiro Hida, Nobuo Shinohara, Hideyoshi Harashima
CANCER MEDICINE, 3, 4, 825, 834, Aug. 2014, [Peer-reviewed]
English, Scientific journal

A nanocarrier system for the delivery of nucleic acids targeted to a pancreatic beta cell line
Yuma Yamada, Mai Tabata, Yukari Yasuzaki, Masatoshi Nomura, Atsushi Shibata, Yuta Ibayashi, Yosuke Taniguchi, Shigeki Sasaki, Hideyoshi Harashima
BIOMATERIALS, 35, 24, 6430, 6438, Aug. 2014, [Peer-reviewed]
English, Scientific journal

An aptamer ligand based liposomal nanocarrier system that targets tumor endothelial cells
Mst Naznin Ara, Takashi Matsuda, Mamoru Hyodo, Yu Sakurai, Hiroto Hatakeyama, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima
BIOMATERIALS, 35, 25, 7110, 7120, Aug. 2014, [Peer-reviewed]
English, Scientific journal

Octaarginine-Modified Liposomes Enhance Cross-Presentation by Promoting the C-Terminal Trimming of Antigen Peptide
Takashi Nakamura, Kouhei Ono, Yoshiteru Suzuki, Rumiko Moriguchi, Kentaro Kogure, Hideyoshi Harashima
MOLECULAR PHARMACEUTICS, 11, 8, 2787, 2795, Aug. 2014, [Peer-reviewed]
English, Scientific journal

RNA Aptamers for Targeting Mitochondria Using a Mitochondria-Based SELEX Method
Yuri Tawaraya, Mamoru Hyodo, Mst Naznin Ara, Yuma Yamada, Hideyoshi Harashima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 37, 8, 1411, 1415, Aug. 2014, [Peer-reviewed]
English, Scientific journal

プラスミドDNA特異的ヒストン修飾による導入遺伝子発現上昇　　　　　　　　　　　　　　　
西原 実香, 神田 元紀, 山門 振一郎, 原島 秀吉, 紙谷 浩之
日本DDS学会学術集会プログラム予稿集, 30回, 165, 165, 日本DDS学会, Jul. 2014
Japanese

Mono-cationic detergents play a critical role in the development of liposome-based gene vector via controlling its lamellarity
Ryosuke Suzuki, Yuma Yamada, Eriko Kawamura, Hideyoshi Harashima
JOURNAL OF NANOPARTICLE RESEARCH, 16, 8, Jul. 2014, [Peer-reviewed]
English, Scientific journal

A new adjuvant delivery system 'cyclic di-GMP/YSK05 liposome' for cancer immunotherapy
Hiroko Miyabe, Mamoru Hyodo, Takashi Nakamura, Yusuke Sato, Yoshihiro Hayakawa, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 184, 20, 27, Jun. 2014, [Peer-reviewed]
English, Scientific journal

Glycerol Monomycolate Is a Novel Ligand for the Human, but Not Mouse Macrophage Inducible C-type Lectin, Mincle
Yuki Hattori, Daisuke Morita, Nagatoshi Fujiwara, Daiki Mori, Takashi Nakamura, Hideyoshi Harashima, Sho Yamasaki, Masahiko Sugita
JOURNAL OF BIOLOGICAL CHEMISTRY, 289, 22, 15405, 15412, May 2014, [Peer-reviewed]
English, Scientific journal

Anatomy of plasmid DNAs with anti-silencing elements
Genki N. Kanda, Shiho Miyamoto, Miwako Kobayashi, Ichiro Matsuoka, Hideyoshi Harashima, Hiroyuki Kamiya
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 464, 1-2, 27, 33, Apr. 2014, [Peer-reviewed]
English, Scientific journal

An apolipoprotein E modified liposomal nanoparticle: Ligand dependent efficiency as a siRNA delivery carrier for mouse-derived brain endothelial cells
Mina Tamaru, Hidetaka Akita, Kazuaki Kajimoto, Yusuke Sato, Hiroto Hatakeyama, Hideyoshi Harashima
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 465, 1-2, 77, 82, Apr. 2014, [Peer-reviewed]
English, Scientific journal

In vivo therapeutic potential of Dicer-hunting siRNAs targeting infectious hepatitis C virus.
Tsunamasa Watanabe, Hiroto Hatakeyama, Chiho Matsuda-Yasui, Yusuke Sato, Masayuki Sudoh, Asako Takagi, Yuichi Hirata, Takahiro Ohtsuki, Masaaki Arai, Kazuaki Inoue, Hideyoshi Harashima, Michinori Kohara
SCIENTIFIC REPORTS, 4, 4750, Apr. 2014, [Peer-reviewed]
English, Scientific journal

Hepatic Monoacylglycerol O-acyltransferase 1 as a Promising Therapeutic Target for Steatosis, Obesity, and Type 2 Diabetes
Yasuhiro Hayashi, Erina Suemitsu, Kazuaki Kajimoto, Yusuke Sato, Afsana Akhter, Yu Sakurai, Hiroto Hatakeyama, Mamoru Hyodo, Noritada Kaji, Yoshinobu Baba, Hideyoshi Harashima
MOLECULAR THERAPY-NUCLEIC ACIDS, 3, e154, Mar. 2014, [Peer-reviewed]
English, Scientific journal

Effects of Silencing the RET/PTC1 Oncogene in Papillary Thyroid Carcinoma by siRNA-Squalene Nanoparticles With and Without Fusogenic Companion GALA-Cholesterol
Hafiz Muhammad Ali, Andrei Maksimenko, Giorgia Urbinati, Hubert Chapuis, Mouna Raouane, Didier Desmaele, Hayashi Yasuhiro, Hideyoshi Harashima, Patrick Couvreur, Liliane Massaad-Massade
THYROID, 24, 2, 327, 338, Feb. 2014, [Peer-reviewed]
English, Scientific journal

Neutral biodegradable lipid-envelope-type nanoparticle using vitamin A-Scaffold for nuclear targeting of plasmid DNA
Hiroki Tanaka, Hidetaka Akita, Ryohei Ishiba, Kota Tange, Masaya Arai, Kazuhiro Kubo, Hideyoshi Harashima
BIOMATERIALS, 35, 5, 1755, 1761, Feb. 2014, [Peer-reviewed]
English, Scientific journal

Nanoparticulation of BCG-CWS for application to bladder cancer therapy
Takashi Nakamura, Masafumi Fukiage, Megumi Higuchi, Akihiro Nakaya, Ikuya Yano, Jun Miyazaki, Hiroyuki Nishiyama, Hideyuki Akaza, Toshihiro Ito, Hiroyuki Hosokawa, Toshinori Nakayama, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 176, 44, 53, Feb. 2014, [Peer-reviewed]
English, Scientific journal

"Semmelweis" in the Innovative Drug Discovery and Developments
Harashima Hideyoshi
Journal of Pharmaceutical Science and Technology, Japan, 74, 1, 1, 1, The Academy of Pharmaceutical Science and Technology, Japan, 2014
Japanese

The initiatives for regulatory science researches of nanomedicines
Harashima Hideyoshi, Hidetaka Akita, Kumiko Sakai-Kato, Takehiko Ishii, Yasuhiro Matsumura, Kazunori Kataoka
Drug Delivery System, 29, 3, 217, 225, Japan Society of Drug Delivery System, 2014, [Peer-reviewed]
Japanese, Scientific journal

Application of apolipoprotein E-modified liposomal nanoparticles as a carrier for delivering DNA and nucleic acid in the brain
Mina Tamaru, Hidetaka Akita, Taichi Nakatani, Kazuaki Kajimoto, Yusuke Sato, Hiroto Hatakeyama, Hideyoshi Harashima
INTERNATIONAL JOURNAL OF NANOMEDICINE, 9, 4267, 4276, 2014, [Peer-reviewed]
English, Scientific journal

Neutralized nanoparticle composed of SS-cleavable and pH-activated lipid-like material as a long-lasting and liver-specific gene delivery system
Masami Ukawa, Hidetaka Akita, Yasuhiro Hayashi, Ryohei Ishiba, Kota Tange, Masaya Arai, Kazuhiro Kubo, Yuriko Higuchi, Kazunori Shimizu, Satoshi Konishi, Mitsuru Hashida, Hideyoshi Harashima
Advanced Healthcare Materials, 3, 8, 1222, 1229, Wiley-VCH Verlag, 2014, [Peer-reviewed]
English, Scientific journal

Suppression of short tract gene conversion in episomal DNA by p53 reduction
Hiroyuki Kamiya, Tetsuya Suzuki, Hideyoshi Harashima
Genes and Environment, 36, 2, 65, 68, The Japanese Environmental Mutagen Society, 2014, [Peer-reviewed]
English, Scientific journal

RNAi-mediated gene knockdown and anti-angiogenic therapy of RCCs using a cyclic RGD-modified liposomal-siRNA system
Yu Sakurai, Hiroto Hatakeyama, Yusuke Sato, Mamoru Hyodo, Hidetaka Akita, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 173, 110, 118, Jan. 2014, [Peer-reviewed]
English, Scientific journal

Cancer multidrug resistance: mechanisms involved and strategies for circumvention using a drug delivery system
Golam Kibria, Hiroto Hatakeyama, Hideyoshi Harashima
ARCHIVES OF PHARMACAL RESEARCH, 37, 1, 4, 15, Jan. 2014, [Peer-reviewed]
English

The systemic administration of an anti-miRNA oligonucleotide encapsulated pH-sensitive liposome results in reduced level of hepatic microRNA-122 in mice
Hiroto Hatakeyama, Manami Murata, Yusuke Sato, Mayumi Takahashi, Noriaki Minakawa, Akira Matsuda, Hideyoshi Harashima
Journal of Controlled Release, 173, 1, 43, 50, 2014, [Peer-reviewed]
English, Scientific journal

The systemic administration of an anti-miRNA oligonucleotide encapsulated pH-sensitive liposome results in reduced level of hepatic microRNA-122 in mice
Hiroto Hatakeyama, Manami Murata, Yusuke Sato, Mayumi Takahashi, Noriaki Minakawa, Akira Matsuda, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 173, 43, 50, Jan. 2014, [Peer-reviewed]
English, Scientific journal

A method for screening mitochondrial fusogenic envelopes for use in mitochondrial drug delivery
Yuma Yamada, Hideyoshi Harashima
Methods in Molecular Biology, 1141, 57, 66, Humana Press Inc., 2014, [Peer-reviewed]
English, Scientific journal

Cytoprotective role of the fatty acid binding protein 4 against oxidative and endoplasmic reticulum stress in 3T3-L1 adipocytes
Kazuaki Kajimoto, Yoshitaka Minami, Hideyoshi Harashima
FEBS OPEN BIO, 4, 602, 610, 2014, [Peer-reviewed]
English, Scientific journal

Application of apolipoprotein E-modified liposomal nanoparticles as a carrier for delivering DNA and nucleic acid in the brain.
Tamaru M, Akita H, Nakatani T, Kajimoto K, Sato Y, Hatakeyama H, Harashima H
International journal of nanomedicine, 9, 4267, 4276, 2014, [Peer-reviewed]

Multifunctional Enveloped Nanodevices (MENDs)
Yusuke Sato, Takashi Nakamura, Yuma Yamada, Hidetaka Akita, Hideyoshi Harashima
NONVIRAL VECTORS FOR GENE THERAPY LIPID- AND POLYMER-BASED GENE TRANSFER, 88, 139, 204, 2014, [Peer-reviewed]
English, In book

In vitro optimization of 2′-OMe-4′-thioribonucleoside-modified anti-microRNA oligonucleotides and its targeting delivery to mouse liver using a liposomal nanoparticle
Mayumi Takahashi, Naoki Yamada, Hiroto Hatakeyama, Manami Murata, Yusuke Sato, Noriaki Minakawa, Hideyoshi Harashima, Akira Matsuda
Nucleic Acids Research, 41, 22, 10659, 10667, Dec. 2013, [Peer-reviewed]
English, Scientific journal

In vitro optimization of 2 '-OMe-4 '-thioribonucleoside-modified anti-microRNA oligonucleotides and its targeting delivery to mouse liver using a liposomal nanoparticle
Mayumi Takahashi, Naoki Yamada, Hiroto Hatakeyama, Manami Murata, Yusuke Sato, Noriaki Minakawa, Hideyoshi Harashima, Akira Matsuda
NUCLEIC ACIDS RESEARCH, 41, 22, 10659, 10667, Dec. 2013, [Peer-reviewed]
English, Scientific journal

Reduced Plasma Glucose by Asparagine Synthetase Knockdown in the Mouse Liver
Shinya Iida, Hiroyuki Kamiya, Akihiro Nakaya, Yasuhiro Hayashi, Akihiro Sawada, Noritada Kaji, Yoshinobu Baba, Hideyoshi Harashima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 36, 12, 2009, 2011, Dec. 2013, [Peer-reviewed]
English, Scientific journal

Intracellular observation of nanocarriers modified with a mitochondrial targeting signal peptide
Eriko Kawamura, Yuma Yamada, Yukari Yasuzaki, Mamoru Hyodo, Hideyoshi Harashima
JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 116, 5, 634, 637, Nov. 2013, [Peer-reviewed]
English, Scientific journal

Development of a Novel Nanoparticle by Dual Modification With the Pluripotential Cell-Penetrating Peptide PepFect6 for Cellular Uptake, Endosomal Escape, and Decondensation of an siRNA Core Complex
Asako Mitsueda, Yuri Shimatani, Masahiro Ito, Takashi Ohgita, Asako Yamada, Susumu Hama, Astrid Graslund, Staffan Lindberg, Ulo Langel, Hideyoshi Harashima, Ikuhiko Nakase, Shiroh Futaki, Kentaro Kogure
BIOPOLYMERS, 100, 6, 698, 704, Nov. 2013, [Peer-reviewed]
English, Scientific journal

A liposomal delivery system that targets liver endothelial cells based on a new peptide motif present in the ApoB-100 sequence.
Akhter A, Hayashi Y, Sakurai Y, Ohga N, Hida K, Harashima H
International journal of pharmaceutics, 456, 1, 195, 201, Elsevier science bv, Nov. 2013, [Peer-reviewed]
English, Liver dysfunction is associated with a variety of liver diseases, including viral or alcoholic hepatitis, fibrosis, cirrhosis, and portal hypertension. A targeted drug delivery system would be very useful in the treatment of these diseases. We herein describe the development of a system comprised of a new peptide-lipid conjugate for the efficient delivery of molecules to LEC. The RLTRKRGLK sequence (3359-3367), which mediates the association of LDL with arterial CSPG and an LDL receptor, was utilized as a ligand for achieving this goal. The peptide modified PEG-LPs (RLTR-PEG-LPs) were efficiently taken up by primary liver endothelial cells (liver ECs) and other types of cells. In vivo biodistribution and confocal microscopy analysis showed that RLTR-PEG-LPs became widely accumulated in LECs within a short time. Distribution of RLTR-PEG-LPs was greatly reduced with a pretreatment of unlabeled RLTR-PEG-LPs, not cationic LPs, indicating that the sequence is important for LECs. The findings indicate that a reverse sequence of RLTR (KLGR) modified PEG-LPs (KLGR-PEG-LP) did the same pattern compared with RLTR-PEG- LPs, suggesting that the RKR or RXXR sequence might be essential for LECs targeting. Collectively RLTR-PEG-LPs and KLGR-PEG-LPs have the potential for delivering drugs to LECs. (C) 2013 Elsevier B.V. All rights reserved.

A DNA microarray-based analysis of immune-stimulatory and transcriptional responses of dendritic cells to KALA-modified nanoparticles
Hidetaka Akita, Soichiro Ishii, Naoya Miura, Sharif Mohammad Shaheen, Yasuhiro Hayashi, Takashi Nakamura, Noritada Kaji, Yoshinobu Baba, Hideyoshi Harashima
BIOMATERIALS, 34, 35, 8979, 8990, Nov. 2013, [Peer-reviewed]
English, Scientific journal

Mitochondrial targeting functional peptides as potential devices for the mitochondrial delivery of a DF-MITO-Porter
Eriko Kawamura, Yuma Yamada, Hideyoshi Harashima
MITOCHONDRION, 13, 6, 610, 614, Nov. 2013, [Peer-reviewed]
English, Scientific journal

Th1-skewed tissue responses to a mycolyl glycolipid in mycobacteria-infected rhesus macaques
Daisuke Morita, Ayumi Miyamoto, Yuki Hattori, Takaya Komori, Takashi Nakamura, Tatsuhiko Igarashi, Hideyoshi Harashima, Masahiko Sugita
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 441, 1, 108, 113, Nov. 2013, [Peer-reviewed]
English, Scientific journal

Investigation of siRNA Nanoparticle Formation Using Mono-Cationic Detergents and Its Use in Gene Silencing in Human HeLa Cells.
Yamada Y, Suzuki R, Harashima H
Cancers, 5, 4, 1413, 1425, Nov. 2013, [Peer-reviewed]

Production of small nano-sized particles by complex formation between polycations and linearized plasmid DNA at a low pH
Ryohei Togashi, Hidetaka Akita, Hideyoshi Harashima
JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 116, 4, 528, 531, Oct. 2013, [Peer-reviewed]
English, Scientific journal

A comparative study between nanoparticle-targeted therapeutics and bioconjugates as obesity medication
Md. Nazir Hossen, Kazuaki Kajimoto, Hidetaka Akita, Mamoru Hyodo, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 171, 2, 104, 112, Oct. 2013, [Peer-reviewed]
English, Scientific journal

Enhancement in selective mitochondrial association by direct modification of a mitochondrial targeting signal peptide on a liposomal based nanocarrier
Yuma Yamada, Hideyoshi Harashima
Mitochondrion, 13, 5, 526, 532, Sep. 2013, [Peer-reviewed]
English, Scientific journal

An Approach to Transgene Expression in Liver Endothelial Cells Using a Liposome-Based Gene Vector Coated with Hyaluronic Acid
Yuma Yamada, Masahiro Hashida, Yasuhiro Hayashi, Mai Tabata, Mamoru Hyodo, Mst Naznin Ara, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima
JOURNAL OF PHARMACEUTICAL SCIENCES, 102, 9, 3119, 3127, Sep. 2013, [Peer-reviewed]
English, Scientific journal

Lipid Envelope-Type Nanoparticle Incorporating a Multifunctional Peptide for Systemic siRNA Delivery to the Pulmonary Endothelium
Kenji Kusumoto, Hidetaka Akita, Taichi Ishitsuka, Yu Matsumoto, Takahiro Nomoto, Ryo Furukawa, Ayman El-Sayed, Hiroto Hatakeyama, Kazuaki Kajimoto, Yuma Yamada, Kazunori Kataoka, Hideyoshi Harashima
ACS NANO, 7, 9, 7534, 7541, Sep. 2013, [Peer-reviewed]
English, Scientific journal

A Neutral envelope-type nanoparticle containing pH-responsive and ss-cleavable lipid-like material as a carrier for plasmid DNA
Hidetaka Akita, Ryohei Ishiba, Hiroto Hatakeyama, Hiroki Tanaka, Yusuke Sato, Kota Tange, Masaya Arai, Kazuhiro Kubo, Hideyoshi Harashima
Advanced Healthcare Materials, 2, 8, 1120, 1125, Aug. 2013, [Peer-reviewed]
English, Scientific journal

The effect of liposomal size on the targeted delivery of doxorubicin to Integrin alpha v beta 3-expressing tumor endothelial cells
Golam Kibria, Hiroto Hatakeyama, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima
BIOMATERIALS, 34, 22, 5617, 5627, Jul. 2013, [Peer-reviewed]
English, Scientific journal

Endocytosis of Gene Delivery Vectors: From Clathrin-dependent to Lipid Raft-mediated Endocytosis
Ayman El-Sayed, Hideyoshi Harashima
MOLECULAR THERAPY, 21, 6, 1118, 1130, Jun. 2013, [Peer-reviewed]
English

The polyethyleneglycol dilemma: Advantage and disadvantage of PEGylation of liposomes for systemic genes and nucleic acids delivery to tumors
Hiroto Hatakeyama, Hidetaka Akita, Hideyoshi Harashima
Biological and Pharmaceutical Bulletin, 36, 6, 892, 899, Jun. 2013, [Peer-reviewed]
English

Correlation between transgen expression and plasmid DNA loss in mouse liver
Ryohei Togashi, Hideyoshi Harashima, Hiroyuki Kamiya
Journal of Gene Medicine, 15, 6-7, 242, 248, Jun. 2013, [Peer-reviewed]
English, Scientific journal

Antiangiogenic nanotherapy for the control of obesity
Kazuaki Kajimoto, Md. Nazir Hossen, Hideyoshi Harashima
NANOMEDICINE, 8, 5, 671, 673, May 2013, [Peer-reviewed]
English

Therapeutic Assessment of Cytochrome C for the Prevention of Obesity Through Endothelial Cell-targeted Nanoparticulate System
Md. Nazir Hossen, Kazuaki Kajimoto, Hidetaka Akita, Mamoru Hyodo, Taichi Ishitsuka, Hideyoshi Harashima
MOLECULAR THERAPY, 21, 3, 533, 541, Mar. 2013, [Peer-reviewed]
English, Scientific journal

Mitochondrial delivery of bongkrekic acid using a MITO-porter prevents the induction of apoptosis in human hela cells
Yuma Yamada, Kohei Nakamura, Ryo Furukawa, Eriko Kawamura, Takuya Moriwaki, Kenji Matsumoto, Katsuhiro Okuda, Mitsuru Shindo, Hideyoshi Harashima
JOURNAL OF PHARMACEUTICAL SCIENCES, 102, 3, 1008, 1015, Mar. 2013, [Peer-reviewed]
English, Scientific journal

Enhanced transgene expression from chromatinized plasmid DNA in mouse liver
Hiroyuki Kamiya, Shiho Miyamoto, Hitomi Goto, Genki N. Kanda, Miwako Kobayashi, Ichiro Matsuoka, Hideyoshi Harashima
International Journal of Pharmaceutics, 441, 1-2, 146, 150, 30 Jan. 2013, [Peer-reviewed]
English, Scientific journal

Major T Cell Response to a Mycolyl Glycolipid Is Mediated by CD1c Molecules in Rhesus Macaques
Daisuke Morita, Yuki Hattori, Takashi Nakamura, Tatsuhiko Igarashi, Hideyoshi Harashima, Masahiko Sugita
INFECTION AND IMMUNITY, 81, 1, 311, 316, Jan. 2013, [Peer-reviewed]
English, Scientific journal

The transfection activity of R8-modified nanoparticles and siRNA condensation using pH sensitive stearylated-octahistidine
Naoyuki Toriyabe, Yasuhiro Hayashi, Hideyoshi Harashima
Biomaterials, 34, 4, 1337, 1343, Jan. 2013, [Peer-reviewed]
English, Scientific journal

Incorporation of polyinosine-polycytidylic acid enhances cytotoxic T cell activity and antitumor effects by octaarginine-modified liposomes encapsulating antigen, but not by octaarginine-modified antigen complex
Takashi Nakamura, Rumiko Moriguchi, Kentaro Kogure, Hideyoshi Harashima
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 441, 1-2, 476, 481, Jan. 2013, [Peer-reviewed]
English, Scientific journal

Particle tracking analysis for the intracellular trafficking of nanoparticles modified with african swine fever virus protein p54-derived peptide
Hidetaka Akita, Kaoru Enoto, Hiroki Tanaka, Hideyoshi Harashima
Molecular Therapy, 21, 2, 309, 317, Nature Publishing Group, 2013, [Peer-reviewed]
English, Scientific journal

Gene silencing via RNAi and siRNA quantification in tumor tissue using MEND, a liposomal siRNA delivery system
Yu Sakurai, Hiroto Hatakeyama, Yusuke Sato, Mamoru Hyodo, Hidetaka Akita, Hideyoshi Harashima
Molecular Therapy, 21, 6, 1195, 1203, Nature Publishing Group, 2013, [Peer-reviewed]
English, Scientific journal

The nanoparticulation by octaarginine-modified liposome improves α-galactosylceramide-mediated antitumor therapy via systemic administration
Takashi Nakamura, Daiki Yamazaki, Jun Yamauchi, Hideyoshi Harashima
Journal of Controlled Release, 171, 2, 216, 224, Elsevier B.V., 2013, [Peer-reviewed]
English, Scientific journal

Efficacy of DHMEQ, a NF-κB inhibitor, in islet transplantation: I. HMGB1 suppression by DHMEQ prevents early islet graft damage
Daisuke Kuraya, Masaaki Watanabe, Yasuyuki Koshizuka, Masaomi Ogura, Tadashi Yoshida, Yoh Asahi, Hirofumi Kamachi, Takashi Nakamura, Hideyoshi Harashima, Michitaka Ozaki, Kazuo Umezawa, Michiaki Matsushita, Kenichiro Yamashita, Satoru Todo
Transplantation, 96, 5, 445, 453, 2013, [Peer-reviewed]
English, Scientific journal

Localization of exogenous DNA to mitochondria in skeletal muscle following hydrodynamic limb vein injection
Yukari Yasuzaki, Yuma Yamada, Tsutomu Kanefuji, Hideyoshi Harashima
Journal of Controlled Release, 172, 3, 805, 811, 2013, [Peer-reviewed]
English, Scientific journal

Development of an Efficient Short Interference RNA (siRNA) Delivery System with a New pH-Sensitive Cationic Lipid
Yusuke Sato, Hiroto Hatakeyama, Mamoru Hyodo, Hidetaka Akita, Hideyoshi Harashima
YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, 132, 12, 1355, 1363, Dec. 2012, [Peer-reviewed]
Japanese

MITO-Porter; A Cutting-edge Technology for Mitochondrial Gene Therapy
Ryo Furukawa, Yuma Yamada, Hideyoshi Harashima
YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, 132, 12, 1389, 1398, Dec. 2012, [Peer-reviewed]
Japanese

Development of a Novel DNA Aptamer Ligand Targeting to Primary Cultured Tumor Endothelial Cells by a Cell-Based SELEX Method
Mst. Naznin Ara, Mamoru Hyodo, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima
PLOS ONE, 7, 12, e50174, Dec. 2012, [Peer-reviewed]
English, Scientific journal

Neuronal injury induces microglial production of macrophage inflammatory protein-1a in rat corticostriatal slice cultures
Toshiyuki Okamura, Takahiro Katayama, Chihiro Obinata, Yukina Iso, Yuzuho Chiba, Hayato Kobayashi, Yuma Yamada, Hideyoshi Harashima, Masabumi Minami
JOURNAL OF NEUROSCIENCE RESEARCH, 90, 11, 2127, 2133, Nov. 2012, [Peer-reviewed]
English, Scientific journal

A pH-sensitive cationic lipid facilitates the delivery of liposomal siRNA and gene silencing activity in vitro and in vivo
Yusuke Sato, Hiroto Hatakeyama, Yu Sakurai, Mamoru Hyodo, Hidetaka Akita, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 163, 3, 267, 276, Nov. 2012, [Peer-reviewed]
English, Scientific journal

RNA Interference-Based Silencing Reveals the Regulatory Role of Fatty Acid-Binding Protein 4 in the Production of IL-6 and Vascular Endothelial Growth Factor in 3T3-L1 Adipocytes
Kazuaki Kajimoto, Shiharu Takayanagi, Shun Sasaki, Hidetaka Akita, Hideyoshi Harashima
ENDOCRINOLOGY, 153, 11, 5629, 5636, Nov. 2012, [Peer-reviewed]
English, Scientific journal

Vascular-targeted nanotherapy for obesity: Unexpected passive targeting mechanism to obese fat for the enhancement of active drug delivery
Md Nazir Hossen, Kazuaki Kajimoto, Hidetaka Akita, Mamoru Hyodo, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 163, 2, 101, 110, Oct. 2012, [Peer-reviewed]
English, Scientific journal

Targeting Mitochondria: Innovation from Mitochondrial Drug Delivery System (DDS) to Mitochondrial Medicine
Yuma Yamada, Hideyoshi Harashima
YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, 132, 10, 1111, 1118, Oct. 2012, [Peer-reviewed]
Japanese

Effects of Endogenous Proteins and microRNA Target Sequence in a Positive Feedback System
Genki N. Kanda, Ryohei Togashi, Hideyoshi Harashima, Hiroyuki Kamiya
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 35, 9, 1534, 1538, Sep. 2012, [Peer-reviewed]
English, Scientific journal

A designed curved DNA sequence remarkably enhances transgene expression from plasmid DNA in mouse liver
S. Fukunaga, G. Kanda, J. Tanase, H. Harashima, T. Ohyama, H. Kamiya
Gene Therapy, 19, 8, 828, 835, Nature Publishing, Aug. 2012, [Peer-reviewed]
English, Scientific journal

Non-linear pharmacokinetics of octaarginine-modified lipid nanoparticles: Barriers from in vitro to in vivo
Yasuhiro Hayashi, Yuki Noguchi, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 161, 3, 757, 762, Aug. 2012, [Peer-reviewed]
English, Scientific journal

Intracellular stability of 2′-OMe-4′-thioribonucleoside modified siRNA leads to long-term RNAi effect
Mayumi Takahashi, Chisato Nagai, Hiroto Hatakeyama, Noriaki Minakawa, Hideyoshi Harashima, Akira Matsuda
Nucleic Acids Research, 40, 12, 5787, 5793, Jul. 2012, [Peer-reviewed]
English, Scientific journal

Efficient Intracellular Delivery of Nucleic Acid Pharmaceuticals Using Cell-Penetrating Peptides
Ikuhiko Nakase, Hidetaka Akita, Kentaro Kogure, Astrid Graslund, Ulo Langel, Hideyoshi Harashima, Shiroh Futaki
ACCOUNTS OF CHEMICAL RESEARCH, 45, 7, 1132, 1139, Jul. 2012, [Peer-reviewed]
English

クロマチン化プラスミドDNAによる発現上昇　　　　　　　　　　　　　　　
紙谷 浩之, 宮本 紫甫, 後藤 仁美, 神田 元紀, 小林 三和子, 松岡 一郎, 原島 秀吉
日本DDS学会学術集会プログラム予稿集, 28回, 163, 163, 日本DDS学会, Jun. 2012
Japanese

Fabrication of Functionalized Double-Lamellar Multifunctional Envelope-Type Nanodevices Using a Microfluidic Chip with a Chaotic Mixer Array
Katsuma Kitazoe, Yeon-Su Park, Noritada Kaji, Yukihiro Okamoto, Manabu Tokeshi, Kentaro Kogure, Hideyoshi Harashima, Yoshinobu Baba
PLOS ONE, 7, 6, e39057, Jun. 2012, [Peer-reviewed]
English, Scientific journal

Post-nuclear gene delivery events for transgene expression by biocleavable polyrotaxanes
Yuma Yamada, Taku Nomura, Hideyoshi Harashima, Atsushi Yamashita, Nobuhiko Yui
Biomaterials, 33, 15, 3952, 3958, May 2012, [Peer-reviewed]
English, Scientific journal

Post-nuclear gene delivery events for transgene expression by biocleavable polyrotaxanes
Yuma Yamada, Taku Nomura, Hideyoshi Harashima, Atsushi Yamashita, Nobuhiko Yui
BIOMATERIALS, 33, 15, 3952, 3958, May 2012, [Peer-reviewed]
English, Scientific journal

FRET from Quantum Dots to Photodecompose Undesired Acceptors and Report the Condensation and Decondensation of Plasmid DNA
Vasudevanpillai Biju, Abdulaziz Anas, Hidetaka Akita, Edakkattuparambil Sidharthan Shibu, Tamitake Itoh, Hideyoshi Harashima, Mitsuru Ishikawa
ACS NANO, 6, 5, 3776, 3788, May 2012, [Peer-reviewed]
English, Scientific journal

Efficient Intradermal Delivery of Superoxide Dismutase Using a Combination of Liposomes and Iontophoresis for Protection against UV-Induced Skin Damage
Kaoru Kigasawa, Moeko Miyashita, Kazuaki Kajimoto, Kiyoshi Kanamura, Hideyoshi Harashima, Kentaro Kogure
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 35, 5, 781, 785, May 2012, [Peer-reviewed]
English, Scientific journal

Different mechanisms for nanoparticle formation between pDNA and siRNA using polyrotaxane as the polycation
Yuma Yamada, Masahiro Hashida, Taku Nomura, Hideyoshi Harashima, Yuichi Yamasaki, Kazunori Kataoka, Atsushi Yamashita, Ryo Katoono, Nobuhiko Yui
ChemPhysChem, 13, 5, 1161, 1165, 10 Apr. 2012, [Peer-reviewed]
English, Scientific journal

1H, 13C and 15N NMR assignments of the Escherichia coli Orf135 protein
Kumiko Kawasaki, Momoko Yoneyama, Naoko Murata-Kamiya, Hideyoshi Harashima, Chojiro Kojima, Yutaka Ito, Hiroyuki Kamiya, Masaki Mishima
Biomol NMR Assign, 6, 1, 1-4, Apr. 2012, [Peer-reviewed]
English, 大腸菌Orf135蛋白質のNMR信号帰属

Different Mechanisms for Nanoparticle Formation between pDNA and siRNA Using Polyrotaxane as the Polycation
Yuma Yamada, Masahiro Hashida, Taku Nomura, Hideyoshi Harashima, Yuichi Yamasaki, Kazunori Kataoka, Atsushi Yamashita, Ryo Katoono, Nobuhiko Yui
CHEMPHYSCHEM, 13, 5, 1161, 1165, Apr. 2012, [Peer-reviewed]
English, Scientific journal

Effect of the Anchor in Polyethylene Glycol-Lipids on the Transfection Activity of PEGylated Cationic Liposomes Encapsulating DNA
Kenji Kusumoto, Hidetaka Akita, Ayman El-Sayed, Hideyoshi Harashima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 35, 4, 445, 448, Apr. 2012, [Peer-reviewed]
English, Scientific journal

Stearylated INF7 peptide enhances endosomal escape and gene expression of PEGylated nanoparticles both in vitro and in vivo
Ayman El-Sayed, Tomoya Masuda, Hidetaka Akita, Hideyoshi Harashima
JOURNAL OF PHARMACEUTICAL SCIENCES, 101, 2, 879, 882, Feb. 2012, [Peer-reviewed]
English, Scientific journal

Delivery of bioactive molecules to the mitochondrial genome using a membrane-fusing, liposome-based carrier, DF-MITO-Porter
Yuma Yamada, Hideyoshi Harashima
BIOMATERIALS, 33, 5, 1589, 1595, Feb. 2012, [Peer-reviewed]
English, Scientific journal

High throughput purification device for gene delivery multifunctional envelope-type nanodevice
Daisuke Shigenaka, Masami Ukawa, Noritada Kaji, Yukihiro Okamoto, Manabu Tokeshi, Hidetaka Akita, Hideyoshi Harashima, Yoshinobu Baba
Proceedings of the 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2012, 1546, 1548, 2012, [Peer-reviewed]
International conference proceedings

The manner in which DNA is packaged with TFAM has an impact on transcription activation and inhibition
Ryo Furukawa, Yuma Yamada, Yuichi Matsushima, Yu-ichi Goto, Hideyoshi Harashima
FEBS OPEN BIO, 2, 145, 150, 2012, [Peer-reviewed]
English, Scientific journal

CREB-binding protein transcription activation domain for enhanced transgene expression by a positive feedback system
Genki Kanda, Hiroshi Ochiai, Hideyoshi Harashima, Hiroyuki Kamiya
JOURNAL OF BIOTECHNOLOGY, 157, 1, 7, 11, Jan. 2012, [Peer-reviewed]
English, Scientific journal

MULTIFUNCTIONAL ENVELOPE-TYPE NANO DEVICE (MEND) FOR ORGANELLE TARGETING VIA A STEPWISE MEMBRANE FUSION PROCESS
Yuma Yamada, Hidetaka Akita, Hideyoshi Harashima
NANOMEDICINE: INFECTIOUS DISEASES, IMMUNOTHERAPY, DIAGNOSTICS, ANTIFIBROTICS, TOXICOLOGY AND GENE MEDICINE, 509, 301, 326, 2012, [Peer-reviewed]
English, In book

Octaarginine- and pH sensitive fusogenic peptide-modified nanoparticles for liver gene delivery
Ikramy A. Khalil, Yasuhiro Hayashi, Ryoichi Mizuno, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 156, 3, 374, 380, Dec. 2011, [Peer-reviewed]
English, Scientific journal

The liposome-incorporating cell wall skeleton of Mycobacterium bovis bacillus Calmette-Guein can directly enhance the susceptibility of cancer cells to lymphokine-activated killer cells through up-regulation of natural-killer group 2, member D ligands
Jun Miyazaki, Koji Kawai, Takahiro Kojima, Takehiro Oikawa, Akira Joraku, Toru Shimazui, Akihiro Nakaya, Ikuya Yano, Takashi Nakamura, Hideyoshi Harashima, Hideyuki Akaza
BJU INTERNATIONAL, 108, 9, 1520, 1526, Nov. 2011, [Peer-reviewed]
English, Scientific journal

Diverse substrate recognition and hydrolysis mechanisms of human NUDT5
Takao Arimori, Haruhiko Tamaoki, Teruya Nakamura, Hiroyuki Kamiya, Shinji Ikemizu, Yasumitsu Takagi, Toru Ishibashi, Hideyoshi Harashima, Mutsuo Sekiguchi, Yuriko Yamagata
NUCLEIC ACIDS RESEARCH, 39, 20, 8972, 8983, Nov. 2011, [Peer-reviewed]
English, Scientific journal

Effects of insulator cHS4 on transgene expression from plasmid DNA in a positive feedback system
Hiroshi Ochiai, Hideyoshi Harashima, Hiroyuki Kamiya
JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 112, 5, 432, 434, Nov. 2011, [Peer-reviewed]
English, Scientific journal

Delivery of nucleic acids and gene delivery　　　　　　　　　　　　　　　
H. Akita, H. Hatakeyama, I. A. Khalil, Y. Yamada, H. Harashima
Comprehensive Biomaterials, 4, 411, 444, Elsevier, Oct. 2011
English, In book

Synthesis, Structure, and Biological Activity of Dumbbell-Shaped Nanocircular RNAs for RNA Interference
Naoko Abe, Hiroshi Abe, Chisato Nagai, Mitsuru Harada, Hiroto Hatakeyama, Hideyoshi Harashima, Takahito Ohshiro, Mizuki Nishihara, Kazuhiro Furukawa, Mizuo Maeda, Satoshi Tsuneda, Yoshihiro Ito
BIOCONJUGATE CHEMISTRY, 22, 10, 2082, 2092, Oct. 2011, [Peer-reviewed]
English, Scientific journal

Mutagenicity of secondary oxidation products of 8-oxo-7,8-dihydro-2 '-deoxyguanosine 5 '-triphosphate (8-hydroxy-2 '- deoxyguanosine 5 '-triphosphate)
Mika Hori, Tetsuya Suzuki, Noriaki Minakawa, Akira Matsuda, Hideyoshi Harashima, Hiroyuki Kamiya
MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS, 714, 1-2, 11, 16, Sep. 2011, [Peer-reviewed]
English, Scientific journal

Correction of Frameshift Mutations with Tailed Duplex DNAs
Yukiko Morita, Hiroyuki Tsuchiya, Hideyoshi Harashima, Hiroyuki Kamiya
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 34, 9, 1465, 1468, Sep. 2011, [Peer-reviewed]
English, Scientific journal

KALA-modified multi-layered nanoparticles as gene carriers for MHC class-I mediated antigen presentation for a DNA vaccine
Sharif M. Shaheen, Hidetaka Akita, Takashi Nakamura, Shota Takayama, Shiroh Futaki, Atsushi Yamashita, Ryo Katoono, Nobuhiko Yui, Hideyoshi Harashima
Biomaterials, 32, 26, 6342, 6350, Sep. 2011, [Peer-reviewed]
English, Scientific journal

KALA-modified multi-layered nanoparticles as gene carriers for MHC class-I mediated antigen presentation for a DNA vaccine
Sharif M. Shaheen, Hidetaka Akita, Takashi Nakamura, Shota Takayama, Shiroh Futaki, Atsushi Yamashita, Ryo Katoono, Nobuhiko Yui, Hideyoshi Harashima
BIOMATERIALS, 32, 26, 6342, 6350, Sep. 2011, [Peer-reviewed]
English, Scientific journal

A20 Silencing by Lipid Envelope-Type Nanoparticles Enhances the Efficiency of Lipopolysaccharide-Activated Dendritic Cells
Shota Warashina, Takashi Nakamura, Hideyoshi Harashima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 34, 8, 1348, 1351, Aug. 2011, [Peer-reviewed]
English, Scientific journal

Endosomal escape and the knockdown efficiency of liposomal-siRNA by the fusogenic peptide shGALA
Yu Sakurai, Hiroto Hatakeyama, Yusuke Sato, Hidetaka Akita, Kentaro Takayama, Sachiko Kobayashi, Shiroh Futaki, Hideyoshi Harashima
BIOMATERIALS, 32, 24, 5733, 5742, Aug. 2011, [Peer-reviewed]
English, Scientific journal

Synthesis and Evaluation of Stearylated Hyaluronic Acid for the Active Delivery of Liposomes to Liver Endothelial Cells
Naoyuki Toriyabe, Yasuhiro Hayashi, Mamoru Hyodo, Hideyoshi Harashima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 34, 7, 1084, 1089, Jul. 2011, [Peer-reviewed]
English, Scientific journal

Improving in Vivo Hepatic Transfection Activity by Controlling Intracellular Trafficking: The Function of GALA and Maltotriose
Hidetaka Akita, Tomoya Masuda, Takashi Nishio, Kenichi Niikura, Kuniharu Ijiro, Hideyoshi Harashima
MOLECULAR PHARMACEUTICS, 8, 4, 1436, 1442, Jul. 2011, [Peer-reviewed]
English, Scientific journal

The Therapeutic Effects of R8-Liposome-BCG-CWS on BBN-Induced Rat Urinary Bladder Carcinoma
Jun Miyazaki, Hiroyuki Nishiyama, Ikuya Yano, Akihiro Nakaya, Hideyasu Kohama, Koji Kawai, Akira Joraku, Takashi Nakamura, Hideyoshi Harashima, Hideyuki Akaza
ANTICANCER RESEARCH, 31, 6, 2065, 2071, Jun. 2011, [Peer-reviewed]
English, Scientific journal

Systemic delivery of siRNA to tumors using a lipid nanoparticle containing a tumor-specific cleavable PEG-lipid
Hiroto Hatakeyama, Hidetaka Akita, Erika Ito, Yasuhiro Hayashi, Motoi Oishi, Yukio Nagasaki, Radostin Danev, Kuniaki Nagayama, Noritada Kaji, Hiroshi Kikuchi, Yoshinobu Baba, Hideyoshi Harashima
BIOMATERIALS, 32, 18, 4306, 4316, Jun. 2011, [Peer-reviewed]
English, Scientific journal

Glycerol monomycolate, a latent tuberculosis-associated mycobacterial lipid, induces eosinophilic hypersensitivity responses in guinea pigs
Yuki Hattori, Isamu Matsunaga, Takaya Komori, Tetsuo Urakawa, Takashi Nakamura, Nagatoshi Fujiwara, Kenji Hiromatsu, Hideyoshi Harashima, Masahiko Sugita
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 409, 2, 304, 307, Jun. 2011, [Peer-reviewed]
English, Scientific journal

A Microbial Glycolipid Functions as a New Class of Target Antigen for Delayed-type Hypersensitivity
Takaya Komori, Takashi Nakamura, Isamu Matsunaga, Daisuke Morita, Yuki Hattori, Hirotaka Kuwata, Nagatoshi Fujiwara, Kenji Hiromatsu, Hideyoshi Harashima, Masahiko Sugita
JOURNAL OF BIOLOGICAL CHEMISTRY, 286, 19, 16800, 16806, May 2011, [Peer-reviewed]
English, Scientific journal

Quantitative analysis of condensation/decondensation status of pDNA in the nuclear sub-domains by QD-FRET
Sharif M. Shaheen, Hidetaka Akita, Atsushi Yamashita, Ryo Katoono, Nobuhiko Yui, Vasudevanpillai Biju, Mitsuru Ishikawa, Hideyoshi Harashima
Nucleic Acids Research, 39, 7, e48, e48, Apr. 2011, [Peer-reviewed]
English, Scientific journal

Quantitative analysis of condensation/decondensation status of pDNA in the nuclear sub-domains by QD-FRET
Sharif M. Shaheen, Hidetaka Akita, Atsushi Yamashita, Ryo Katoono, Nobuhiko Yui, Vasudevanpillai Biju, Mitsuru Ishikawa, Hideyoshi Harashima
NUCLEIC ACIDS RESEARCH, 39, 7, E48, U108, Apr. 2011, [Peer-reviewed]
English, Scientific journal

Topology of octaarginines (R8) or IRQ ligand on liposomes affects the contribution of macropinocytosis- and caveolae-mediated cellular uptake
Diky Mudhakir, Hidetaka Akita, Hideyoshi Harashima
REACTIVE & FUNCTIONAL POLYMERS, 71, 3, 340, 343, Mar. 2011, [Peer-reviewed]
English, Scientific journal

A multifunctional envelope type nano device (MEND) for gene delivery to tumours based on the EPR effect: A strategy for overcoming the PEG dilemma
Hiroto Hatakeyama, Hidetaka Akita, Hideyoshi Harashima
ADVANCED DRUG DELIVERY REVIEWS, 63, 3, 152, 160, Mar. 2011, [Peer-reviewed]
English

Noninvasive and efficient transdermal delivery of CpG-oligodeoxynucleotide for cancer immunotherapy
Kaoru Kigasawa, Kazuaki Kajimoto, Takashi Nakamura, Susumu Hama, Kiyoshi Kanamura, Hideyoshi Harashima, Kentaro Kogure
JOURNAL OF CONTROLLED RELEASE, 150, 3, 256, 265, Mar. 2011, [Peer-reviewed]
English, Scientific journal

Modulation of Endosomal Escape of IRQ-PEGylated Nano-carrier
Diky Mudhakir, Hidetaka Akita, Hideyoshi Harashima
4TH NANOSCIENCE AND NANOTECHNOLOGY SYMPOSIUM (NNS2011): AN INTERNATIONAL SYMPOSIUM, 1415, 2011, [Peer-reviewed]
English, International conference proceedings

Design of Smart Nano Device for Intracellular Targeting
Diky Mudhakir, Erdal Tan, Hidetaka Akita, Hideyoshi Harashima
4TH NANOSCIENCE AND NANOTECHNOLOGY SYMPOSIUM (NNS2011): AN INTERNATIONAL SYMPOSIUM, 1415, 2011, [Peer-reviewed]
English, International conference proceedings

Octaarginine-modified liposomes enhance the anti-oxidant effect of Lecithinized superoxide dismutase by increasing its cellular uptake
Ryo Furukawa, Yuma Yamada, Mitsuko Takenaga, Rie Igarashi, Hideyoshi Harashima
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 404, 3, 796, 801, Jan. 2011, [Peer-reviewed]
English, Scientific journal

Noninvasive and persistent transfollicular drug delivery system using a combination of liposomes and iontophoresis
Kazuaki Kajimoto, Masahiko Yamamoto, Misuzu Watanabe, Kaoru Kigasawa, Kiyoshi Kanamura, Hideyoshi Harashima, Kentaro Kogure
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 403, 1-2, 57, 65, Jan. 2011, [Peer-reviewed]
English, Scientific journal

A touch-and-go lipid wrapping technique in microfluidic channels for rapid fabrication of multifunctional envelope-type gene delivery nanodevices
Katsuma Kitazoe, Jun Wang, Noritada Kaji, Yukihiro Okamoto, Manabu Tokeshi, Kentaro Kogure, Hideyoshi Harashima, Yoshinobu Baba
LAB ON A CHIP, 11, 19, 3256, 3262, 2011, [Peer-reviewed]
English, Scientific journal

Reprint of: Nanoparticles for ex vivo siRNA delivery to dendritic cells for cancer vaccines: Programmed endosomal escape and dissociation
Hidetaka Akita, Kentaro Kogure, Rumiko Moriguchi, Yoshio Nakamura, Tomoko Higashi, Takashi Nakamura, Satoshi Serada, Minoru Fujimoto, Tetsuji Naka, Shiroh Futaki, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 149, 1, 58, 64, Jan. 2011, [Peer-reviewed]
English

Special Issue: Recent Advancements of DDS research in japan
Hideyoshi Harashima, Akihiko Kikuchi, Kazunori Kataoka
JOURNAL OF CONTROLLED RELEASE, 149, 1, 1, 1, Jan. 2011, [Peer-reviewed]
English, Scientific journal

(Review) Application of a fusiogenic peptide GALA for intracellular delivery　　　　　　　　　　　　　　　
Ikuhiko Nakase, Kentaro Kogure, Hideyoshi Harashima, Shiroh Futaki
Methods in Molecular Biology, 683, 525, 533, 2011, [Peer-reviewed]

内在性転写活性化因子を用いた細胞系における外来遺伝子発現の活性化　　　　　　　　　　　　　　　
神田 元紀, 落合 浩史, 原島 秀吉, 紙谷 浩之
日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 83回・33回, 1P, 1247, (公社)日本生化学会, Dec. 2010
Japanese

2種類の「発現持続型」プラスミドの核内挙動の違い　　　　　　　　　　　　　　　
冨樫 亮平, 神田 元紀, 紙谷 浩之, 原島 秀吉
日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 83回・33回, 1P, 1248, (公社)日本生化学会, Dec. 2010
Japanese

Reporter gene expression at single-cell level characterized with real-time RT-PCR, chemiluminescence, fluorescence, and electrochemical imaging
Hitoshi Shiku, Daisuke Okazaki, Junya Suzuki, Yasufumi Takahashi, Tatsuya Murata, Hidetaka Akita, Hideyoshi Harashima, Kosuke Ino, Tomokazu Matsue
FEBS LETTERS, 584, 18, 4000, 4008, Sep. 2010, [Peer-reviewed]
English, Scientific journal

2-Methacryloyloxyethyl phosphorylcholine polymer (MPC)-coating improves the transfection activity of GALA-modified lipid nanoparticles by assisting the cellular uptake and intracellular dissociation of plasmid DNA in primary hepatocytes
Masami Ukawa, Hidetaka Akita, Tomoya Masuda, Yasuhiro Hayashi, Tomohiro Konno, Kazuhiko Ishihara, Hideyoshi Harashima
BIOMATERIALS, 31, 24, 6355, 6362, Aug. 2010, [Peer-reviewed]
English, Scientific journal

Intranuclear DNA Release Is a Determinant of Transfection Activity for a Non-viral Vector: Biocleavable Polyrotaxane as a Supramolecularly Dissociative Condenser for Efficient Intranuclear DNA Release
Yuma Yamada, Taku Nomura, Hideyoshi Harashima, Atsushi Yamashita, Ryo Katoono, Nobuhiko Yui
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 33, 7, 1218, 1222, Jul. 2010, [Peer-reviewed]
English, Scientific journal

Ornithine and Tryptophan Analogs as Efficient Polycations for Short Interference RNA Delivery to Tumor Cells
Yusuke Sato, Hiroto Hatakeyama, Hideyoshi Harashima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 33, 7, 1246, 1249, Jul. 2010, [Peer-reviewed]
English, Scientific journal

Positive Feedback System Provides Efficient and Persistent Transgene Expression
Hiroshi Ochiai, Hideyoshi Harashima, Hiroyuki Kamiya
MOLECULAR PHARMACEUTICS, 7, 4, 1125, 1132, Jul. 2010, [Peer-reviewed]
English, Scientific journal

Mitochondrial matrix delivery using MITO-Porter, a liposome-based carrier that specifies fusion with mitochondrial membranes
Yukari Yasuzaki, Yuma Yamada, Hideyoshi Harashima
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 397, 2, 181, 186, Jun. 2010, [Peer-reviewed]
English, Scientific journal

Particle Tracking of Intracellular Trafficking of Octaarginine-modified Liposomes: A Comparative Study With Adenovirus
Hidetaka Akita, Kaoru Enoto, Tomoya Masuda, Hiroyuki Mizuguchi, Tomomi Tani, Hideyoshi Harashima
MOLECULAR THERAPY, 18, 5, 955, 964, May 2010, [Peer-reviewed]
English, Scientific journal

Effects of base excision repair proteins on mutagenesis by 8-oxo-7,8-dihydroguanine (8-hydroxyguanine) paired with cytosine and adenine
Tetsuya Suzuki, Hideyoshi Harashima, Hiroyuki Kamiya
DNA REPAIR, 9, 5, 542, 550, May 2010, [Peer-reviewed]
English, Scientific journal

Nanoparticles for ex vivo siRNA delivery to dendritic cells for cancer vaccines: Programmed endosomal escape and dissociation
Hidetaka Akita, Kentaro Kogure, Rumiko Moriguchi, Yoshio Nakamura, Tomoko Higashi, Takashi Nakamura, Satoshi Serada, Minoru Fujimoto, Tetsuji Naka, Shiroh Futaki, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 143, 3, 311, 317, May 2010, [Peer-reviewed]
English, Scientific journal

Roles of specialized DNA polymerases in mutagenesis by 8-hydroxyguanine in human cells
Hiroyuki Kamiya, Ayaka Yamaguchi, Tetsuya Suzuki, Hideyoshi Harashima
MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS, 686, 1-2, 90, 95, Apr. 2010, [Peer-reviewed]
English, Scientific journal

Leptin-derived peptide, a targeting ligand for mouse brain-derived endothelial cells via macropinocytosis
Mina Tamaru, Hidetaka Akita, Takahiro Fujiwara, Kazuaki Kajimoto, Hideyoshi Harashima
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 394, 3, 587, 592, Apr. 2010, [Peer-reviewed]
English, Scientific journal

Targeted sequence alteration of a chromosomal locus in mouse liver
Hiroyuki Kamiya, Masayuki Uchiyama, Jingshu Piao, Yoshimichi Nakatsu, Teruhisa Tsuzuki, Hideyoshi Harashima
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 387, 1-2, 180, 183, Mar. 2010
English, Scientific journal

An Improvement Delivery of siRNA by IRQ Ligand-Modified Double Membranous Nano-Carrier
Diky Mudhakir, Erdal Tan, Hidetaka Akita, Hideyoshi Harashima
THIRD NANOSCIENCE AND NANOTECHNOLOGY SYMPOSIUM 2010 - NNSB2010, 1284, 199, +, 2010, [Peer-reviewed]
English, International conference proceedings

A Novel Nonviral Gene Delivery System: Multifunctional Envelope-Type Nano Device
Hiroto Hatakeyama, Hidetaka Akita, Kentaro Kogure, Hideyoshi Harashima
NANO/MICRO BIOTECHNOLOGY, 119, 197, +, 2010, [Peer-reviewed]
English, Scientific journal

Intranuclear DNA release is a determinant of transfection activity for a non-viral vector: Biocleavable polyrotaxane as a supramolecularly dissociative condenser for efficient intranuclear DNA release
Yuma Yamada, Taku Nomura, Hideyoshi Harashima, Atsushi Yamashita, Ryo Katoono, Nobuhiko Yui
Biological and Pharmaceutical Bulletin, 33, 7, 1218, 1222, 2010, [Peer-reviewed]
English, Scientific journal

Roles of specialized DNA polymerases in mutagenesis by oxidized guanine.
Hiroyuki Kamiya, Kazuya Satou, Mika Hori, Ayaka Yamaguchi, Hideyoshi Harashima
Nucleic acids symposium series (2004), 53, 223, 224, 01 Dec. 2009

NUDT5 hydrolyzes oxidized deoxyribonucleoside diphosphates with broad substrate specificity
Hiroyuki Kamiya, Mika Hori, Takao Arimori, Mutsuo Sekiguchi, Yuriko Yamagata, Hideyoshi Harashima
DNA REPAIR, 8, 10, 1250, 1254, Oct. 2009
English, Scientific journal

A pH-sensitive fusogenic peptide facilitates endosomal escape and greatly enhances the gene silencing of siRNA-containing nanoparticles in vitro and in vivo
Hiroto Hatakeyama, Erika Ito, Hidetaka Akita, Motoi Oishi, Yukio Nagasaki, Shiroh Futaki, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 139, 2, 127, 132, Oct. 2009, [Peer-reviewed]
English, Scientific journal

Effects of non-B DNA sequences on transgene expression
Hiroyuki Kamiya, Hitomi Goto, Hideyoshi Harashima
JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 108, 1, 18, 21, Jul. 2009
English, Scientific journal

Effects of carriers on transgene expression from plasmids containing a DNA sequence with high histone affinity
Hiroyuki Kamiya, Satoki Fukunaga, Takashi Ohyama, Hideyoshi Harashima
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 376, 1-2, 99, 103, Jul. 2009
English, Scientific journal

Effect of Polyethyleneglycol Spacer on the Binding Properties of Nuclear Localization Signal-Modified Liposomes to Isolated Nucleus
Dai Kurihara, Hidetaka Akita, Asako Kudo, Tomoya Masuda, Shiroh Futaki, Hideyoshi Harashima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 32, 7, 1303, 1306, Jul. 2009, [Peer-reviewed]
English, Scientific journal

Involvement of specialized DNA polymerases in mutagenesis by 8-hydroxy-dGTP in human cells
Kazuya Satou, Mika Hori, Kazuaki Kawai, Hiroshi Kasai, Hideyoshi Harashima, Hiroyuki Kamiya
DNA REPAIR, 8, 5, 637, 642, May 2009
English, Scientific journal

A Novel Technique To Investigate the Decondensation of pc-DNA3.1-luc in Nuclear Sub-Domain with QD-FRET
Sharif M. Shaheen, Hidetaka Akita, Atsushi Yamashita, Ryo Katoono, Nobuhiko Yui, Vasudevanpillai Biju, Mitsuru Ishikawa, Hideyoshi Harashima
MOLECULAR THERAPY, 17, S330, S330, May 2009, [Peer-reviewed]
English

Multi-layered nanoparticles for penetrating the endosome and nuclear membrane via a step-wise membrane fusion process
Hidetaka Akita, Asako Kudo, Arisa Minoura, Masaya Yamaguti, Ikrarny A. Khalil, Rumiko Moriguchi, Tomoya Masuda, Radostin Danev, Kuniaki Nagayama, Kentaro Kogure, Hideyoshi Harashima
BIOMATERIALS, 30, 15, 2940, 2949, May 2009, [Peer-reviewed]
English, Scientific journal

Development of Efficient siRNA Delivery System to Tumor Cells by Combining Octaarginine, GALA and Enzymatically-Cleavable PEG-Lipid
Yu Sakurai, Hiroto Hatakeyama, Hidetaka Akita, Motoi Oishi, Yukio Nagasaki, Shiro Futaki, Hideyoshi Harashima
MOLECULAR THERAPY, 17, Suppl. 1, S388, S389, May 2009, [Peer-reviewed]
English, Scientific journal

A pH-Sensitive Fusogenic Peptide Facilitates Endosomal Escape and Greatly Enhanced the Gene Silencing of siRNA-Containing Nanoparticles In Vitro and In Vivo
Hiroto Hatakeyama, Erika Itho, Hidetaka Akita, Motoi Oihi, Yukio Nagasaki, Shiroh Futaki, Hideyoshi Harashima
MOLECULAR THERAPY, 17, S388, S388, May 2009, [Peer-reviewed]
English

Immunoprotection against murine bladder carcinoma by octaarginine-modified liposomes incorporating cell wall of Mycobacterium bovis bacillus Calmette-Guérin.
Joraku A, Homhuan A, Kawai K, Yamamoto T, Miyazaki J, Kogure K, Yano I, Harashima H, Akaza H
BJU international, 103, 5, 686, 693, Mar. 2009, [Peer-reviewed]

Delivery of Macromolecules Using Arginine-Rich Cell-Penetrating Peptides: Ways to Overcome Endosomal Entrapment
Ayman El-Sayed, Shiroh Futaki, Hideyoshi Harashima
AAPS JOURNAL, 11, 1, 13, 22, Mar. 2009, [Peer-reviewed]
English

Non-linear pharmacodynamics in the transfection efficiency of a non-viral gene delivery system
Rumiko Moriguchi, Kentaro Kogure, Hideyoshi Harashima
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 363, 1-2, 192, 198, Nov. 2008, [Peer-reviewed]
English, Scientific journal

Oligosaccharide-Mediated Nuclear Transport of Nanoparticles
Kenichi Niikura, Shota Sekiguchi, Takashi Nishio, Tomoya Masuda, Hidetaka Akita, Yasutaka Matsuo, Kentaro Kogure, Hideyoshi Harashima, Kuniharu Ijiro
CHEMBIOCHEM, 9, 16, 2623, 2627, Nov. 2008, [Peer-reviewed]
English, Scientific journal

Supramolecular control of polyplex dissociation and cell transfection: efficacy of amino groups and threading cyclodextrins in biocleavable polyrotaxanes.
Atsushi Yamashita, Daizo Kanda, Ryo Katoono, Nobuhiko Yui, Tooru Ooya, Atsushi Maruyama, Hidetaka Akita, Kentaro Kogure, Hideyoshi Harashima
J Control Release, 131, 2, 137, 144, Oct. 2008, [Peer-reviewed]
English, Scientific journal, A novel strategy for gene delivery using biocleavable polyrotaxanes, in which dimethylaminoethyl-modified alpha-cyclodextrins (DMAE-alpha-CDs) are threaded onto a poly(ethylene glycol) (PEG) chain capped with benzyloxycarbonyl-L-tyrosine via disulfide linkages (DMAE-SS-PRX), involves the formation of a stable polyion complex (polyplex) against a counter polyanion and the intracellular plasmid DNA (pDNA) release from the polyplex accompanied by the supramolecular dissociation of DMAE-SS-PRXs. In this study, we prepared biocleavable polyrotaxanes with different numbers of threading alpha-CD and amino (DMAE) groups to enhance the transfection activity of DMAE-SS-PRXs. 29DMAE-alpha18-SS-PRX, in which the numbers of alpha-CD molecules and amino groups were 18 and 29 respectively, exhibited a high transfection activity compared with other PRXs. The transfection activity of DMAE-SS-PRXs seems to be related to the efficacy of pDNA release from those polyplexes, which was controlled by the number of alpha-CD and/or amino groups in the polyrotaxane carrier. Most of the DMAE-SS-PRX polyplexes released the pDNA only in the presence of both 10 mM DTT and of the counter-polyanion, as expected, e

Mitochondrial drug delivery systems for macromolecule and their therapeutic application to mitochondrial diseases
Yuma Yamada, Hideyoshi Harashima
ADVANCED DRUG DELIVERY REVIEWS, 60, 13-14, 1439, 1462, Oct. 2008, [Peer-reviewed]
English

Effects of target sequence and sense versus anti-sense strands on gene correction with single-stranded DNA fragments
Hiroyuki Kamiya, Masayuki Uchiyama, Yoshimichi Nakatsu, Teruhisa Tsuzuki, Hideyoshi Harashima
JOURNAL OF BIOCHEMISTRY, 144, 4, 431, 436, Oct. 2008
English, Scientific journal

Supramolecular control of polyplex dissociation and cell transfection: Efficacy of amino groups and threading cyclodextrins in biocleavable polyrotaxanes
Atsushi Yamashita, Daizo Kanda, Ryo Katoono, Nobuhiko Yui, Tooru Ooya, Atsushi Maruyama, Hidetaka Akita, Kentaro Kogure, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 131, 2, 137, 144, Oct. 2008, [Peer-reviewed]
English, Scientific journal

Efficient MHC class I presentation by controlled intracellular trafficking of antigens in octaarginine-modified liposomes
Takashi Nakamura, Rumiko Moriguchi, Kentaro Kogure, Nilabh Shastri, Hideyoshi Harashima
MOLECULAR THERAPY, 16, 8, 1507, 1514, Aug. 2008, [Peer-reviewed]
English, Scientific journal

Advances in non-viral gene delivery: using multifunctional envelope-type nano-device
Hidetaka Akita, Hideyoshi Harashima
EXPERT OPINION ON DRUG DELIVERY, 5, 8, 847, 859, Aug. 2008, [Peer-reviewed]
English

Photosensitized breakage and damage of DNA by CdSe-ZnS quantum dots
AbdulAziz Anas, Hidetaka Akita, Hideyoshi Harashima, Tamitake Itoh, Mitsuru Ishikawa, Vasudevanpillai Biju
JOURNAL OF PHYSICAL CHEMISTRY B, 112, 32, 10005, 10011, Aug. 2008, [Peer-reviewed]
English, Scientific journal

Improved gene correction efficiency with a tailed duplex DNA fragment
Hiroyuki Tsuchiya, Masayuki Uchiyarna, Kazuhiro Hara, Yoshimichi Nakatsu, Teruhisa Tsuzuki, Hideo Inoue, Hideyoshi Harashima, Hiroyuki Kamiya
BIOCHEMISTRY, 47, 33, 8754, 8759, Aug. 2008
English, Scientific journal

On-chip fabrication of mutifunctional envelope-type nanodevices for gene delivery
Hiroshi Kuramoto, Yeon-Su Park, Noritada Kaji, Manabu Tokeshi, Kentaro Kogure, Yasuo Shinohara, Hideyoshi Harashima, Yoshinobu Baba
ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 391, 8, 2729, 2733, Aug. 2008
English, Scientific journal

An artificial virus-like nano carrier system: enhanced endosomal escape of nanoparticles via synergistic action of pH-sensitive fusogenic peptide derivatives
Kentaro Sasaki, Kentaro Kogure, Shinji Chaki, Yoshio Nakamura, Rumiko Moriguchi, Hirofumi Hamada, Radostin Danev, Kuniaki Nagayama, Shiroh Futaki, Hideyoshi Harashima
ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 391, 8, 2717, 2727, Aug. 2008, [Peer-reviewed]
English, Scientific journal

Octaarginine- and octalysine-modified nanoparticles have different modes of endosomal escape
Ayman El-Sayed, Ikramy A. Khalil, Kentaro Kogure, Shiroh Futaki, Hideyoshi Harashima
JOURNAL OF BIOLOGICAL CHEMISTRY, 283, 34, 23450, 23461, Aug. 2008, [Peer-reviewed]
English, Scientific journal

Cellular attachment and internalization of cationic liposomes containing mycobacterial cell wall
Atthachai Homhuan, Hideyoshi Harashima, Ikuya Yano
SCIENCEASIA, 34, 2, 179, 185, Jun. 2008, [Peer-reviewed]
English, Scientific journal

Octaarginine-modified liposomes: Enhanced cellular uptake and controlled intracellular trafficking
Krarny A. Khalil, Kentaro Kogure, Shiroh Futaki, Hideyoshi Harashima
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 354, 1-2, 39, 48, Apr. 2008, [Peer-reviewed]
English

Multifunctional envelope-type nano device (MEND) as a non-viral gene delivery system
Kentaro Kogure, Hidetaka Akita, Yuma Yamada, Hideyoshi Harashima
ADVANCED DRUG DELIVERY REVIEWS, 60, 4-5, 559, 571, Mar. 2008, [Peer-reviewed]
English

Specificity of mutations induced by incorporation of oxidized dNTPs into DNA by human DNA polymerase eta.
Katsuhiko Hidaka, Masami Yamada, Hiroyuki Kamiya, Chikahide Masutani, Hideyoshi Harashima, Fumio Hanaoka, Takehiko Nohmi
DNA repair, 7, 3, 497, 506, 01 Mar. 2008, [Peer-reviewed], [International Magazine]
English, Aberrant oxidation is a property of many tumor cells. Oxidation of DNA precursors, i.e., deoxynucleotide triphosphates (dNTPs), as well as DNA is a major cause of genome instability. Here, we report that human DNA polymerase eta (h Poleta) incorporates oxidized dNTPs, i.e., 2-hydroxy-2'-deoxyadenosine 5'-triphosphate (2-OH-dATP) and 8-hydroxy-2'-deoxyguanosine 5'-triphosphate (8-OH-dGTP), into DNA in an erroneous and efficient manner, thereby inducing various types of mutations during in vitro gap-filling DNA synthesis. When 2-OH-dATP was present at a concentration equal to those of the four normal dNTPs in the reaction mixture, DNA synthesis by h Poleta enhanced the frequency of G-to-T transversions eight-fold higher than that of the transversions in control where only the normal dNTPs were present. When 8-OH-dGTP was present at an equimolar concentration to the normal dNTPs, it enhanced the frequency of A-to-C transversions 17-fold higher than the control. It also increased the frequency of C-to-A transversions about two-fold. These results suggest that h Poleta incorporates 2-OH-dATP opposite template G and incorporates 8-OH-dGTP opposite template A and slightly opposite template C during DNA synthesis. Besides base substitutions, h Poleta enhanced the frequency of single-base frameshifts and deletions with the size of more than 100 base pairs when 8-OH-dGTP was present in the reaction mixture. Since h Poleta is present in replication foci even without exogenous DNA damage, we suggest that h Poleta may be involved in induction of various types of mutations through the erroneous and efficient incorporation of oxidized dNTPs into DNA in human cells.

Programmed Packaging: A New Drug Delivery System and its Application to Gene Therapy
Kentaro Kogure, Hidetaka Akita, Hiroyuki Kamiya, Hideyoshi Harashima
Modern Biopharmaceuticals: Design, Development and Optimization, 4, 1521, 1536, Wiley-VCH Verlag GmbH, 05 Feb. 2008
English, In book

MITO-Porter: A liposome-based carrier system for delivery of macromolecules into mitochondria via membrane fusion.
Yuma Yamada, Hidetaka Akita, Hiroyuki Kamiya, Kentaro Kogure, Takenori Yamamoto, Yasuo Shinohara, Kikuji Yamashita, Hideo Kobayashi, Hiroshi Kikuchi, Hideyoshi Harashima
Biochimica et biophysica acta, 1778, 2, 423, 32, Feb. 2008, [International Magazine]
English, Scientific journal, Mitochondria are the principal producers of energy in higher cells. Mitochondrial dysfunction is implicated in a variety of human diseases, including cancer and neurodegenerative disorders. Effective medical therapies for such diseases will ultimately require targeted delivery of therapeutic proteins or nucleic acids to the mitochondria, which will be achieved through innovations in the nanotechnology of intracellular trafficking. Here we describe a liposome-based carrier that delivers its macromolecular cargo to the mitochondrial interior via membrane fusion. These liposome particles, which we call MITO-Porters, carry octaarginine surface modifications to stimulate their entry into cells as intact vesicles (via macropinocytosis). We identified lipid compositions for the MITO-Porter which promote both its fusion with the mitochondrial membrane and the release of its cargo to the intra-mitochondrial compartment in living cells. Thus, the MITO-Porter holds promise as an efficacious system for the delivery of both large and small therapeutic molecules into mitochondria.

Development of lipid particles targeted via sugar-lipid conjugates as novel nuclear gene delivery system
Tomoya Masuda, Hidetaka Akita, Takashi Nishio, Kenichi Niikura, Kentaro Kogure, Kuniharu Ijiro, Hideyoshi Harashima
BIOMATERIALS, 29, 6, 709, 723, Feb. 2008, [Peer-reviewed]
English, Scientific journal

Nonviral gene delivery
Hidetaka Akita, Hideyoshi Harashima
GENE THERAPY FOR RENAL DISEASES AND TRANSPLANTATION, 159, 13, 29, 2008, [Peer-reviewed]
English, Scientific journal

Effect of surface-modification with octaarginine versus octalysine on cellular uptake and endosomal escape of nanoparticles
Ayman El-Sayed, Ikramy A. Khalil, Kentaro Kogure, Shiroh Futaki, Hideyoshi Harashima
YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, 128, 111, 112, 2008, [Peer-reviewed]
English

Supramolecular control of polyplex dissociation and cell transfection: Efficacy of amine and threading cyclodextrin in biocleavable polyrotaxanes
Yamashita A, Kanda D, Katoono R, Yui N, Ooya T, Maruyama A, Akita H, Kogure K, Harashima H
8th World Biomaterials Congress 2008, 4, 2008, [Peer-reviewed]

Base excision repair system suppresses mutagenesis caused by 8-hydroxy-dGTP in Escherichia coli.
Tetsuya Suzuki, Kazuo Yamamoto, Hideyoshi Harashima, Hiroyuki Kamiya
Nucleic acids symposium series (2004), 51, 52, 01 Dec. 2007

UvrAとUvrBは酸化損傷ヌクレオチドにより誘発される変異を促進する
堀 美香, 石黒 智恵子, 鈴木 哲矢, 中川 紀子, 布柴 達男, 倉光 成紀, 山本 和生, 葛西 宏, 原島 秀吉, 紙谷 浩之
日本放射線影響学会大会講演要旨集, 50回, 69, 69, (一社)日本放射線影響学会, Nov. 2007
Japanese

Multifunctional envelope-type nano device for non-viral gene delivery: Concept and application of Programmed Packaging
Kentaro Kogure, Hidetaka Akita, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 122, 3, 246, 251, Oct. 2007, [Peer-reviewed]
English, Scientific journal

Tumor targeting of doxorubicin by anti-MT1-MMP antibody-modified PEG liposomes
Hiroto Hatakeyama, Hidetaka Akita, Emi Ishida, Koichi Hashimoto, Hideo Kobayashi, Takanori Aoki, Junko Yasuda, Kenichi Obata, Hiroshi Kikuchi, Tatsuhiro Ishida, Hiroshi Kiwada, Hideyoshi Harashima
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 342, 1-2, 194, 200, Sep. 2007, [Peer-reviewed]
English, Scientific journal

New packaging method of mycobacterial cell wall using octaarginine-modified liposomes: Enhanced uptake by and immunostimulatory activity of dendritic cells
Atthachal Homhuan, Kentaro Kogure, Hideyuki Akaza, Shiroh Futaki, Takashi Naka, Yukiko Fujita, Ikuya Yano, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 120, 1-2, 60, 69, Jul. 2007, [Peer-reviewed]
English, Scientific journal

2-Hydroxy-2'-deoxyadenosine 5'-triphosphate enhances A.T --> C.G mutations caused by 8-hydroxy-2'-deoxyguanosine 5'-triphosphate by suppressing its degradation upon replication in a HeLa extract.
Kazuya Satou, Hiroshi Kasai, Chikahide Masutani, Fumio Hanaoka, Hideyoshi Harashima, Hiroyuki Kamiya
Biochemistry, 46, 22, 6639, 46, 05 Jun. 2007, [Peer-reviewed], [International Magazine]
English, The coexistence effects of multiple kinds of oxidized deoxyribonucleotides were examined using an SV40 origin-dependent in vitro replication system with a HeLa extract. Oxidized dGTP and dATP, 8-hydroxy-2'-deoxyguanosine 5'-triphosphate (8-OH-dGTP) and 2-hydroxy-2'-deoxyadenosine 5'-triphosphate (2-OH-dATP), were used in this study. The mutation frequency synergistically increased when the two oxidized deoxyribonucleotides were together in the reaction. 2-OH-dATP enhanced the mutagenicity of 8-OH-dGTP, since the induced mutations were A.T --> C.G transversions. The contribution of the highly error-prone DNA polymerase eta was unlikely, since similar results were observed with an XP-V cell extract. The possible involvement of 2-hydroxyadenine in the complementary (template) strand was excluded on the basis of experiments using plasmids containing 2-hydroxyadenine as templates in the reactions with 8-OH-dGTP. 2-OH-dATP suppressed hydrolysis of 8-OH-dGTP, suggesting that the inhibition of the MTH1 protein played the major role in the enhancement. These results highlight the importance of specific hydrolysis of 8-OH-dGTP for the suppression of its induced mutation.

Octaarginine-modified multifunctional envelope-type nano device for siRNA
Yoshio Nakamura, Kentaro Kogure, Shiroh Futaki, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 119, 3, 360, 367, Jun. 2007, [Peer-reviewed]
English, Scientific journal

Efficient and erroneous incorporation of oxidized DNA precursors by human DNA polymerase eta.
Masatomi Shimizu, Petr Gruz, Hiroyuki Kamiya, Chikahide Masutani, Yan Xu, Yukio Usui, Hiroshi Sugiyama, Hideyoshi Harashima, Fumio Hanaoka, Takehiko Nohmi
Biochemistry, 46, 18, 5515, 22, 08 May 2007, [Peer-reviewed], [International Magazine]
English, Scientific journal, Altered oxidative metabolism is a property of many tumor cells. Oxidation of DNA precursors, i.e., dNTP pool, as well as DNA is a major source of mutagenesis and carcinogenesis. Here, we report the remarkable nature of human DNA polymerase eta that incorporates oxidized dNTPs into a nascent DNA strand in an efficient and erroneous manner. The polymerase almost exclusively incorporated 8-hydroxy-dGTP (8-OH-dGTP) opposite template adenine (A) at 60% efficiency of normal dTTP incorporation, and incorporated 2-hydroxy-dATP (2-OH-dATP) opposite template thymine (T), guanine (G), or cytosine (C) at substantial rates. The synthetic primers having 8-hydroxy-G paired with template A or 2-hydroxy-A paired with template T, G, or C at the termini were efficiently extended. In contrast, human DNA polymerase iota incorporated 8-OH-dGTP opposite template A with much lower efficiency and did not incorporate 2-OH-dATP opposite any of the template bases. It did not extend the primers having the oxidized bases at the termini either. We propose that human DNA polymerase eta may participate in oxidative mutagenesis through the efficient and erroneous incorporation of oxidized dNTPs during DNA synthesis.

The location of the left-handedly curved DNA sequence affects exogenous DNA expression in vivo
Hiroyuki Kamiya, Satoki Fukunaga, Takashi Ohyama, Hideyoshi Harashima
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 461, 1, 7, 12, May 2007
English, Scientific journal

Octaarginine-modified multifunctional envelope-type nanoparticles for gene delivery
I. A. Khalil, K. Kogure, S. Futaki, S. Hama, H. Akita, M. Ueno, H. Kishida, M. Kudoh, Y. Mishina, K. Kataoka, M. Yamada, H. Harashima
GENE THERAPY, 14, 8, 682, 689, Apr. 2007, [Peer-reviewed]
English, Scientific journal

Accumulation of O-GlcNAc-displaying CdTe quantum dots in cells in the presence of ATP
Kenichi Niikura, Takashi Nishio, Hidetaka Akita, Yasutaka Matsuo, Ryosuke Kamitani, Kentaro Kogure, Hideyoshi Harashima, Kuniharu Ijiro
CHEMBIOCHEM, 8, 4, 379, 384, Mar. 2007, [Peer-reviewed]
English, Scientific journal

Effect of transferrin receptor-targeted liposomal doxorubicin in P-glycoprotein-mediated drug resistant tumor cells
Tomotaka Kobayashi, Tatsuhiro Ishida, Yurie Okada, Saori Ise, Hideyoshi Harashima, Hiroshi Kiwada
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 329, 1-2, 94, 102, Feb. 2007, [Peer-reviewed]
English, Scientific journal

A more excellent construction method of a multi functional envelope-type nano device on microdevice for gene therapy
Hiroshi Kuramoto, Noritada Kaji, Kentarou Kogure, Manabu Tokeshi, Yasuo Shinohara, Hideyoshi Harashima, Yoshinobu Baba
Proceedings of the 11th International Conference on Miniaturized Systems for Chemistry and Life Sciences, uTAS 2007, 781, 783, 2007
International conference proceedings

Correlation between the Phosphohydrolase Activity of the Escherichia coli Orf135 (NudG) Protein and Mutation Suppression
Hiroyuki Kamiya, Emiko Iida, Hideyoshi Harashima
Genes and Environment, 29, 2, 63, 66, 2007
English, Scientific journal

Development of a novel systemic gene delivery system for cancer therapy with a tumor-specific cleavable PEG-lipid
H. Hatakeyama, H. Akita, K. Kogure, M. Oishi, Y. Nagasaki, Y. Kihira, M. Ueno, H. Kobayashi, H. Kikuchi, H. Harashima
GENE THERAPY, 14, 1, 68, 77, Jan. 2007, [Peer-reviewed]
English, Scientific journal

Synthesis of a biocleavable polyrotaxane-plasmid DNA (pDNA) polyplex and its use for the rapid nonviral delivery of pDNA to cell nuclei.
Atsushi Yamashita, Nobuhiko Yui, Tooru Ooya, Arihiro Kano, Atsushi Maruyama, Hidetaka Akita, Kentaro Kogure, Hideyoshi Harashima
Nat Protoc, 1, 6, 2861, 2869, Dec. 2006, [Peer-reviewed]
English, Scientific journal, This protocol provides a method for synthesizing a biocleavable polyrotaxane/plasmid DNA (pDNA) polyplex and for using it to deliver pDNA into cell nuclei. The biocleavable polyrotaxane is synthesized in four steps: (i) introduction of disulfide linkages at both terminals of PEG, (ii) preparation of an inclusion complex between disulfide-containing PEG and alpha-cyclodextrins (alpha-CDs), (iii) synthesis of polyrotaxane and (iv) modification of alpha-CDs in the polyrotaxane with dimethylethylenediamine. A polyplex of pDNA with the polyrotaxane is formed when the two compounds are dissolved together in a phosphate buffer. Subcellular localization of rhodamine-labeled pDNA in fluorescently labeled organelles is quantified by Z-series of confocal images captured by confocal laser scanning microscopy. Significant amounts of pDNA delivered to the nucleus can be expected as well as high transfection activity of the polyplex. This protocol can be completed in 23-32 d.

Mutagenic properties of oxidized GTP and ATP in in vitro transcription-reverse transcription.
Hiroyuki Kamiya, Akihiro Suzuki, Kazuaki Kawai, Hiroshi Kasai, Hideyoshi Harashima
Nucleic acids symposium series (2004), 99, 100, 01 Dec. 2006

Positively charged polyethylenimines enhance nasal absorption of the negatively charged drug, low molecular weight heparin
Tianzhi Yang, Alamdar Hussain, Shuhua Bai, Ikramy A. Khalil, Hideyoshi Harashima, Fakhrul Ahsan
JOURNAL OF CONTROLLED RELEASE, 115, 3, 289, 297, Oct. 2006, [Peer-reviewed]
English, Scientific journal

損傷ヌクレオチドによる遺伝情報不安定性誘発とその制御 酸化ヌクレオチド誘発変異とヌクレオチド除去修復酵素UvrABCとの関連　　　　　　　　　　　　　　　
堀 美香, 石黒 智恵子, 中川 紀子, 倉光 成紀, 山本 和生, 葛西 宏, 原島 秀吉, 紙谷 浩之
日本放射線影響学会大会講演要旨集, 49回, 75, 75, (一社)日本放射線影響学会, Sep. 2006
Japanese

Impact of convective flow on the cellular uptake and transfection activity of lipoplex and adenovirus
Takahiro Fujiwara, Hidetaka Akita, Katsuko Furukawa, Takashi Ushida, Hiroyuki Mizuguchi, Hideyoshi Harashima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 29, 7, 1511, 1515, Jul. 2006, [Peer-reviewed]
English, Scientific journal

Involvement of Y-family DNA polymerases in mutagenesis caused by oxidized nucleotides in Escherichia coli
M Yamada, T Nunoshiba, M Shimizu, P Gruz, H Kamiya, H Harashima, T Nohmi
JOURNAL OF BACTERIOLOGY, 188, 13, 4992, 4995, Jul. 2006
English, Scientific journal

Cellular uptake and subsequent intracellular trafficking of R8-liposomes introduced at low temperature.
Akitada Iwasa, Hidetaka Akita, Ikramy Khalil, Kentaro Kogure, Shiroh Futaki, Hideyoshi Harashima
Biochimica et biophysica acta, 1758, 6, 713, 20, Jun. 2006, [International Magazine]
English, Scientific journal, Intracellular trafficking is a determining factor in the transgene expression efficiency of gene vectors. In the present study, the mechanism of the cellular uptake of octaarginine (R8)-modified liposomes, when introduced at 37 degrees C and 4 degrees C, was investigated in living cells. Compared with 37 degrees C, the uptake of R8-liposomes was only slightly reduced at 4 degrees C. Dual imaging of liposomes and plasma membranes revealed that R8-liposomes were internalized by vesicular transport, and partially escaped to the cytosol at the perinuclear region at 37 degrees C. When introduced at 4 degrees C, intracellular liposomes were observed within a specific region close to the plasma membrane, and internalization of the plasma membrane was completely inhibited. Therefore, at 4 degrees C, R8-liposomes appear to enter cells via unique pathway, which is separate and distinct from energy-dependent vesicular transport. The subsequent nuclear delivery of encapsulated pDNA, when introduced at 4 degrees C, was less prominent compared with those introduced at 37 degrees C. Collectively, these findings demonstrate that a vesicular transport-independent pathway is responsible for the cellular uptake of liposomes. In addition, the uptake route is closely related to the subsequent nuclear delivery process; the operation of an endogenous vesicular sorting system is advantageous for the nuclear delivery of pDNA.

Erratum: Size and topology of exogenous DNA as determinant factors of transgene transcription in mammalian cells (Gene Therapy (2002) vol. 9 (1500-1507) 10.1038/sj.gt.3301831)
H. Kamiya, J. Yamazaki, H. Harashima
Gene Therapy, 13, 932, 01 Jun. 2006

Silencing of exogenous DNA in cultured cells
Hiroshi Ochiai, Hideyoshi Harashima, Hiroyuki Kamiya
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 29, 6, 1294, 1296, Jun. 2006
English, Scientific journal

Cellular uptake and subsequent intracellular trafficking of R8-liposomes introduced at low temperature
Akitada Iwasa, Hidetaka Akita, Ikramy Khalil, Kentaro Kogure, Shiroh Futaki, Hideyoshi Harashima
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, 1758, 6, 713, 720, Jun. 2006, [Peer-reviewed]
English, Scientific journal

Significant Silencing Effect of a Multifunctional Envelope-Type Nano Device Encapsulating Antisense Oligodeoxynucleotide and Double-Stranded siRNA
Kentaro Kogure, Yoshio Nakamura, Ikramy A. Khalil, Shiroh Futaki, Hideyoshi Harashima
MOLECULAR THERAPY, 13, S202, S203, May 2006, [Peer-reviewed]
English

Cellular Uptake and Subsequent Intracellular of R8-Liposomes Introduced at Low Temperature
Akitada Iwasa, Hidetaka Akita, Ikramy Khalil, Kentaro Kogure, Shiroh Futaki, Hideyoshi Harashima
MOLECULAR THERAPY, 13, S129, S129, May 2006, [Peer-reviewed]
English

Interaction of hemoglobin vesicles, a cellular-type artificial oxygen carrier, with human plasma: Effects on coagulation, kallikrein-kinin, and complement systems
Hideki Abe, Mitsuhiro Fujihara, Hiroshi Azuma, Hisami Ikeda, Kenji Ikebuchi, Shinji Takeoka, Eishun Tsuchida, Hideyoshi Harashima
Artificial Cells, Blood Substitutes, and Biotechnology, 34, 1, 1, 10, 01 Apr. 2006, [Peer-reviewed]
English, Scientific journal

Quantitative comparison of intracellular trafficking and nuclear transcription between adenoviral and lipoplex systems
S Hama, H Akita, R Ito, H Mizuguchi, T Hayakawa, H Harashima
MOLECULAR THERAPY, 13, 4, 786, 794, Apr. 2006, [Peer-reviewed]
English, Scientific journal

Uptake pathways and subsequent intracellular trafficking in nonviral gene delivery
IA Khalil, K Kogure, H Akita, H Harashima
PHARMACOLOGICAL REVIEWS, 58, 1, 32, 45, Mar. 2006, [Peer-reviewed]
English

Biocleavable polyrotaxane - Plasmid DNA polyplex for enhanced gene delivery
T Ooya, HS Choi, A Yamashita, N Yui, Y Sugaya, A Kano, A Maruyama, H Akita, R Ito, K Kogure, H Harashima
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 128, 12, 3852, 3853, Mar. 2006, [Peer-reviewed]
English, Scientific journal

High density of octaarginine stimulates macropinocytosis leading to efficient intracellular trafficking for gene expression
I.A. Khalil, K. Kogure, S. Futaki, H. Harashima
Journal of Biological Chemistry, 281, 6, 3544, 3551, 10 Feb. 2006, [Peer-reviewed]
English

[Envelope-type nano device for gene delivery].
Kogure K, Akita H, Harashima H
Nihon rinsho. Japanese journal of clinical medicine, 64, 2, 258, 262, Feb. 2006, [Peer-reviewed], [Domestic magazines]
Japanese, Scientific journal, We introduced a new concept "Programmed Packaging" for the rational development of non viral gene delivery system. There are many non-viral gene delivery system, however, their utility is limited in the in vitro experiments. The major reason is considered that it is difficult to package many functional devices into a nanoparticle. The new concept which we introduce here is named as Programmed Packaging, which is composed of the strategy to targeting, molecular design of functional devices to overcome barriers, and assembly technique to package them into a nanoparticle. In this article, we explain this concept and provide newly developed gene delivery systems based on this concept.

Intranuclear disposition of exogenous DNA in vivo: Silencing, methylation and fragmentation
H Ochiai, H Harashima, H Kamiya
FEBS LETTERS, 580, 3, 918, 922, Feb. 2006
English, Scientific journal

Sugar-displaying quantum dots for cellular stress sensing　　　　　　　　　　　　　　　
Kenichi Niikura, Takashi Nishio, Yasutaka Matsuo, Hidetaka Akita, Kentaro Kogure, Hideyoshi Harashima, Kuniharu Ijiro
Polymer Preprints, Japan, 55, 2, 5451, 5452, 2006
Japanese, International conference proceedings

Synthesis of sugar-displaying quantum dots and their intracellular dynamics　　　　　　　　　　　　　　　
Takashi Nishio, Kenichi Niikura, Yasutaka Matsuo, Hidetaka Akita, Kentaro Kogure, Hideyoshi Harashima, Kuniharu Ijiro
Polymer Preprints, Japan, 55, 1, 2145, 2006
Japanese, International conference proceedings

Effects of supramolecular structure of cationic-polyrotaxanes on pDNA compaction
Yamashita A, Ooya T, Kanda D, Choi H.S, Yui N, Maruyama A, Akita H, Kogure K, Harashima H
Polymer Preprints, Japan, 55, 1, 2006, [Peer-reviewed]

Quantitative comparison of intracellular trafficking between artificial and viral vector: Mechanism underlying the difference in dominant rate-limiting process
Hidetaka Akita, Susumu Hama, Hiroyuki Mizuguchi, Hideyoshi Harashima
Progress on Post-Genome Technologies, 521, 525, 2006, [Peer-reviewed]
English, International conference proceedings

Transient expression of Drosophila melanogaster deoxynucleoside kinase gene enhances cytotoxicity of nucleoside analogs
Hiroyuki Kamiya, Hiroshi Ochiai, Hideyoshi Harashima, Mana Ito, Akira Matsuda
NUCLEOSIDES NUCLEOTIDES & NUCLEIC ACIDS, 25, 4-6, 553, 560, 2006
English, Scientific journal

Evaluation of the nuclear delivery and intra-nuclear transcription of plasmid DNA condensed with μ (mu) and NLS-μ by cytoplasmic and nuclear microinjection: A comparative study with poly-L-lysine
H. Akita, M. Tanimoto, T. Masuda, K. Kogure, S. Hama, K. Ninomiya, S. Futaki, H. Harashima
Journal of Gene Medicine, 8, 2, 198, 206, 2006, [Peer-reviewed], [International Magazine]
English, Scientific journal, BACKGROUND: The efficient nuclear delivery of plasmid DNA (pDNA) is essential for the development of a promising non-viral gene vector. In an attempt to achieve nuclear delivery, NLS-mu, a novel pDNA condenser, was prepared. This consists of mu, a highly potent polypeptide for condensing the pDNA, and a SV40 T antigen-derived nuclear localization signal (NLS(SV40)). METHODS: The utility of NLS-mu was assessed in terms of green fluorescent protein (GFP) expression after cytoplasmic and nuclear microinjection of GFP-encoding pDNA along with the transfection, and compared with mu and poly-L-lysine (PLL). Trans-gene expression after cytoplasmic microinjection was affected by the efficiencies of nuclear transfer and following intra-nuclear transcription. To evaluate the nuclear transfer process separately, we introduced a parameter, a nuclear transfer score (NT score), which was calculated as the trans-gene expression after cytoplasmic microinjection divided by that after nuclear microinjection. RESULTS: As expected, the rank order of trans-gene expression after the transfection and cytoplasmic microinjection was NLS-mu > mu > PLL. However, the calculated NT scores were unexpectedly ranked as mu = NLS-mu > PLL, suggesting that mu, and not NLS(SV40), is responsible for the nuclear delivery of pDNA. In addition, confocal images of rhodamine-labeled pDNA indicated that pDNA condensed with mu and NLS-mu was delivered as a condensed form. In comparing the nuclear transcription, the rank order of trans-gene expression after nuclear microinjection was PLL = NLS-mu > mu, suggesting that intra-nuclear transcription is inhibited by efficient condensation by mu, and is avoided by the attachment of NLS(SV40). CONCLUSIONS: Collectively, NLS-mu, which consists of chimeric functions, is an excellent DNA condenser, and the process is based on mu-derived nuclear transfer and NLS(SV40)-derived efficient intra-nuclear transcription.

Pharmacokinetics and brain uptake of lactoferrin in rats
B Ji, A Maeda, M Higuchi, K Inoue, H Akita, H Harashima, T Suhara
LIFE SCIENCES, 78, 8, 851, 855, Jan. 2006, [Peer-reviewed]
English, Scientific journal

In vitro evaluation of cationic polyrotaxanes as gene carrier
Kanda D, Yamashita A, Katoono R, Ooya T, Yui N, Maruyama A, Akita H, Kogure K, Harashima H
Polymer Preprints, Japan, 55, 2, 5367, 5368, 2006, [Peer-reviewed]

Synthesis of a biocleavable polyrotaxane-plasmid DNA (pDNA) polyplex and its use for the rapid nonviral delivery of pDNA to cell nuclei
Atsushi Yamashita, Nobuhiko Yui, Tooru Ooya, Arihiro Kano, Atsushi Maruyama, Hidetaka Akita, Kentaro Kogure, Hideyoshi Harashima
NATURE PROTOCOLS, 1, 6, 2861, 2869, 2006, [Peer-reviewed]
English, Scientific journal

Conversion of nucleotide sequence with single-stranded DNA fragment prepared from phagemid DNA.
Hiroyuki Tsuchiya, Tomoko Sawamura, Masayuki Uchiyama, Hideyoshi Harashima, Hiroyuki Kamiya
Nucleic acids symposium series (2004), 93, 94, 01 Dec. 2005

Factors affecting SFHR gene correction efficiency with single-stranded DNA fragment
H Tsuchiya, H Harashima, H Kamiya
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 336, 4, 1194, 1200, Nov. 2005
English, Scientific journal

Sunflower-shaped cyclodextrin-conjugated poly(-lysine) polyplex as a controlled intracellular trafficking device
H. S. Choi, A. Yamashita, OOYA Tooru, N. Yui, H. Akita, K. Kogure, R. Ito, H. Harashima
ChemPhysChem, Vol. 6, pp. 1986-1990, 11, 1986, 90, Nov. 2005, [Peer-reviewed], [International Magazine]
English, Scientific journal

Unique features of a pH-sensitive fusogenic peptide that improves the transfection efficiency of cationic liposomes
S Futaki, Y Masui, Nakase, I, Y Sugiura, T Nakamura, K Kogure, H Harashima
JOURNAL OF GENE MEDICINE, 7, 11, 1450, 1458, Nov. 2005, [Peer-reviewed]
English, Scientific journal

Effect of methylated adenine in plasmid DNA on transgene expression in mice
H Ochiai, H Harashima, H Kamiya
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 28, 10, 2019, 2022, Oct. 2005
English, Scientific journal

A multifunctional envelope-type nano device for novel gene delivery of siRNA plasmids
R. Moriguchi, K. Kogure, H. Akita, S. Futaki, M. Miyagishi, K. Taira, H. Harashima
International Journal of Pharmaceutics, 301, 1-2, 277, 285, 14 Sep. 2005, [Peer-reviewed], [International Magazine]
English, Scientific journal, A multifunctional envelope-type nano device (MEND) for use in the delivery of siRNA expression plasmids is described. The plasmid DNA encoding anti-luciferase short interfering RNA (siRNA) was condensed by poly-L-lysine (PLL) and packaged into the MEND. The silencing effect of the MEND(PLL) showed a 96% inhibition of luciferase activities in a co-transfection study. The silencing effect was maintained at more than 60%, even under the 100-fold diluted conditions. In the luciferase transformed cells, however, the MEND(PLL) showed no significant silencing effect (10%), indicating heterogeneity in transfection by the MEND(PLL). To solve this problem, the DNA condensing agents were optimized by comparing PLL, stearyl octaarginine (STR-R8) and protamine (Prot). No difference in silencing effect (95-97%) was found among these MENDs in a co-transfection study. However, the MEND(Prot) showed a 70% silencing effect in the transformed cells. These results suggest that the MEND(Prot) has less heterogeneity in transfection, while the MEND(PLL) and the MEND(STR-R8) have large heterogeneities. These results demonstrate that MEND(Prot) is a promising gene delivery system for siRNA expression plasmids with less heterogeneity associated with the transfection.

Mutagenesis induced by oxidized DNA precursors: Roles of Y family DNA polymerases in Escherichia coli
K Satou, M Yamada, T Nohmi, H Harashima, H Kamiya
CHEMICAL RESEARCH IN TOXICOLOGY, 18, 8, 1271, 1278, Aug. 2005
English, Scientific journal

Kinetic analysis of protein production after DNA transfection
Y Yumada, H Kamiya, H Harashima
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 299, 1-2, 34, 40, Aug. 2005
English, Scientific journal

Construction of a multifunctional envelope-type nano device by a SUV*-fusion method
K Sasaki, K Kogure, S Chaki, Y Kihira, M Ueno, H Harashima
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 296, 1-2, 142, 150, May 2005, [Peer-reviewed]
English, Scientific journal

Increased SFHR gene correction efficiency with sense single-stranded DNA
H Tsuchiya, H Harashima, H Kamiya
JOURNAL OF GENE MEDICINE, 7, 4, 486, 493, Apr. 2005
English, Scientific journal

Amino acid residues involved in substrate recognition of the Escherichia coli Orf 135 protein
E Iida, K Satou, M Mishima, C Kojima, H Harashima, H Kamiya
BIOCHEMISTRY, 44, 15, 5683, 5689, Apr. 2005
English, Scientific journal

Improved cell viability of linear polyethyleneimine through -cyclodextrin inclusion for effective gene delivery
A. Yamashita, H. S. Choi, OOYA Tooru, N. Yui, H. Akita, K. Kogure, R. Ito, H. Harashima
ChemPhysChem, Vol. 7, pp. 297-302, 297, 302, Feb. 2005, [Peer-reviewed]
English, Scientific journal

Pharmacokinetics of gene delivery in cells
Hidetaka Akita, Ikramy A. Khalil, Kentaro Kogure, Hideyoshi Harashima
Non-viral Gene Therapy: Gene Design and Delivery, 135, 154, Springer-Verlag Tokyo, 2005, [Peer-reviewed]
English, In book

Octaarginine-mediated stimulation of clathrin-independent endocytic uptake leading to enhanced gene expression through avoiding lysosomal degradation
IA Khalil, K Kogure, S Futaki, H Harashima
YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, 125, 94, 95, 2005, [Peer-reviewed]
English, Scientific journal

Newly designed DNA fragments for gene correction
Hiroyuki Kamiya, Hiroyuki Tsuchiya, Hideyoshi Harashima
Non-viral Gene Therapy: Gene Design and Delivery, 315, 322, Springer-Verlag Tokyo, 2005
English, In book

Dual hydrolysis of diphosphate and triphosphate derivatives of oxidized deoxyadenosine by Orf17 (NtpA), a MutT-type enzyme
M Hori, K Fujikawa, H Kasai, H Harashima, H Kamiya
DNA REPAIR, 4, 1, 33, 39, Jan. 2005
English, Scientific journal

Cytokine induction by a bacterial DNA-specific modified base
H Tsuchiya, T Matsuda, H Harashima, H Kamiya
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 326, 4, 777, 781, Jan. 2005, [Peer-reviewed]
English, Scientific journal

Pharmacokinetic analysis of the tissue distribution of octaarginine modified liposomes in mice.
D. Mudhakir, H. Akita, I.A. Khalil, S. Futaki, H. Harashima
Drug metabolism and pharmacokinetics., 20, 4, 275, 281, 2005, [Peer-reviewed], [International Magazine]
English, Scientific journal, We recently found that octaarginine modified liposomes (R8-Lip) can be efficiently internalized by cultured cells. The purpose of the present study was to quantitatively determine the effect of R8-density on the tissue distribution of R8-Lip in mice, using their clearance as an index. R8 was introduced in the form of stearylated R8 (STR-R8). The liposomes were composed of cholesterol and egg phosphatidylcholine and were labeled with [ (3)H]cholesteryl hexadecyl ether. Various densities of R8 (3%, 10% and 30%) containing liposomes were prepared with a diameter of approximately 70-80 nm. The tissue distribution of R8-Lip was determined after their i.v. administration into mice and the effect of R8-density on tissue distribution was compared with uptake clearance, the calculated tissue distribution divided by the area under the blood concentration-time course. As results, R8-Lip were more rapidly eliminated from circulating blood and distributed to many tissues, especially liver depending on the R8-density. However, the tissue uptake clearance represented similar value to that of positively charge liposomes. Based on these results, we conclude that the R8-dependent increase in R8-Lip in various tissues tested indicates that positive charge, but not PTD function derived from R8 predominantly responsible for the enhancement of tissue distribution. Therefore, it is suggested that topology control of R8 is important to exhibit the PTD function.

Mutagenesis by damaged deoxyribonucleotides and its prevention by MutT-type hydrolyzing enzymes.
Hiroyuki Kamiya, Kazuya Satou, Mika Hori, Emiko Iida, Chieko Ishiguro, Hideyoshi Harashima
Nucleic acids symposium series (2004), 48, 271, 272, 01 Dec. 2004, [International Magazine]
English, Scientific journal

mutT株における8-ヒドロキシ-dGTP誘発変異の上昇 MutT蛋白質の酸化dGTPによる変異の防御の直接的証拠　　　　　　　　　　　　　　　
紙谷 浩之, 石黒 智恵子, 原島 秀吉
日本放射線影響学会大会講演要旨集, 47回, 73, 73, (一社)日本放射線影響学会, Nov. 2004
Japanese

Important amino acids in the phosphohydrolase module of Escherichia coli Orf135
H Kamiya, E Iida, H Harashima
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 323, 3, 1063, 1068, Oct. 2004
English, Scientific journal

Development of a non-viral multifunctional envelope-type nano device by a novel lipid film hydration method
K. Kogure, R. Moriguchi, K. Sasaki, M. Ueno, S. Futaki, H. Harashima
Journal of Controlled Release, 98, 2, 317, 323, 11 Aug. 2004, [Peer-reviewed]
English

Factors governing the in vivo tissue uptake of transferrin-coupled polyethylene glycol liposomes in vivo
H Hatakeyama, H Akita, K Maruyama, T Suhara, H Harashima
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 281, 1-2, 25, 33, Aug. 2004, [Peer-reviewed]
English, Scientific journal

Characterization of a Nudix hydrolase from Deinococcus radiodurans with a marked specificity for (deoxy)ribonucleoside 5′-diphosphates
David I. Fisher, Jared L. Cartwright, Hideyoshi Harashima, Hiroyuki Kamiya, Alexander G. McLennan
BMC Biochemistry, 5, 1, 24, BioMed Central Ltd., 17 May 2004
English, Scientific journal

Transferrin-modified liposomes equipped with a pH-sensitive fusogenic peptide: An artificial viral-like delivery system
T Kakudo, S Chaki, S Futaki, Nakase, I, K Akaji, T Kawakami, K Maruyama, H Kamiya, H Harashima
BIOCHEMISTRY, 43, 19, 5618, 5628, May 2004, [Peer-reviewed]
English, Scientific journal

Mechanism of improved gene transfer by the N-terminal stearylation of octaarginine: enhanced cellular association by hydrophobic core formation
IA Khalil, S Futaki, M Niwa, Y Baba, N Kaji, H Kamiya, H Harashima
GENE THERAPY, 11, 7, 636, 644, Apr. 2004, [Peer-reviewed]
English, Scientific journal

Quantitative three-dimensional analysis of the intracellular trafficking of plasmid DNA transfected by a nonviral gene delivery system using confocal laser scanning microscopy
H. Akita, R. Ito, I.A. Khalil, S. Futaki, H. Harashima
Molecular Therapy, 9, 3, 443, 451, Mar. 2004, [Peer-reviewed]
English

Probing the substrate recognition mechanism of the human MTH1 protein by nucleotide analogs.
Hiroyuki Kamiya, Hiroyuki Yakushiji, Laurence Dugué, Mitsuhide Tanimoto, Sylvie Pochet, Yusaku Nakabeppu, Hideyoshi Harashima
Journal of molecular biology, 336, 4, 843, 50, 27 Feb. 2004, [Peer-reviewed], [International Magazine]
English

An assessment of relative transcriptional availability from nonviral vectors
R Tachibana, N Ide, Y Shinohara, H Harashima, CA Hunt, H Kiwada
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 270, 1-2, 315, 321, Feb. 2004, [Peer-reviewed]
English, Scientific journal

Factors affecting membrane transport of octaarginine-modified liposomes for gene delivery
LA Khalil, K Kogure, S Futaki, H Harashima
YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, 124, 113, 116, 2004, [Peer-reviewed]
English, Scientific journal

Pharmacokinetic modeling of species-dependent enhanced bioavailability of trifluorothymidine by thymidine phosphorylase inhibitor.
Hiroyuki Tsuchiya, Keizou Kuwata, Sekio Nagayama, Kazumasa Yamashita, Hiroyuki Kamiya, Hideyoshi Harashima
Drug metabolism and pharmacokinetics, 19, 206, 215, 01 Jan. 2004

Suppression of spontaneous and hydrogen peroxide-induced mutations by a MutT-type nucleotide pool sanitization enzyme, the Escherichia coli Orf135 protein
H Kamiya, E Iida, N Murata-Kamiya, Y Yamamoto, T Miki, H Harashima
GENES TO CELLS, 8, 12, 941, 950, Dec. 2003
English, Scientific journal

Pharmacokinetic and pharmacodynamic considerations in gene therapy
H Kamiya, H Akita, H Harashima
DRUG DISCOVERY TODAY, 8, 21, 990, 996, Nov. 2003
English

No enhancement of nuclear entry by direct conjugation of a nuclear localization signal peptide to linearized DNA
M Tanimoto, H Kamiya, N Minakawa, A Matsuda, H Harashima
BIOCONJUGATE CHEMISTRY, 14, 6, 1197, 1202, Nov. 2003
English, Scientific journal

The complement- but not mannose receptor-mediated phagocytosis is involved in the hepatic uptake of cetylmannoside-modified liposomes in situ
H Matsuo, K Funato, H Harashima, H Kiwada
JOURNAL OF DRUG TARGETING, 11, 5, 273, 277, Jun. 2003, [Peer-reviewed]
English, Scientific journal

Mutagenic effects of 2-hydroxy-dATP on replication in a HeLa extract: induction of substitution and deletion mutations
K Satou, H Harashima, H Kamiya
NUCLEIC ACIDS RESEARCH, 31, 10, 2570, 2575, May 2003
English, Scientific journal

Erroneous incorporation of oxidized DNA precursors by Y-family DNA polymerases.
Masatomi Shimizu, Petr Gruz, Hiroyuki Kamiya, Su-Ryang Kim, Francesca M Pisani, Chikahide Masutani, Yusuke Kanke, Hideyoshi Harashima, Fumio Hanaoka, Takehiko Nohmi
EMBO reports, 4, 3, 269, 73, Mar. 2003, [Peer-reviewed], [International Magazine]
English, Scientific journal, Deranged oxidative metabolism is a property of many tumour cells. Oxidation of the deoxynucleotide triphosphate (dNTP) pool, as well as DNA, is a major cause of genome instability. Here, we report that two Y-family DNA polymerases of the archaeon Sulfolobus solfataricus strains P1 and P2 incorporate oxidized dNTPs into nascent DNA in an erroneous manner: the polymerases exclusively incorporate 8-OH-dGTP opposite adenine in the template, and incorporate 2-OH-dATP opposite guanine more efficiently than opposite thymine. The rate of extension of the nascent DNA chain following on from these incorporated analogues is only slightly reduced. These DNA polymerases have been shown to bypass a variety of DNA lesions. Thus, our results suggest that the Y-family DNA polymerases promote mutagenesis through the erroneous incorporation of oxidized dNTPs during DNA synthesis, in addition to facilitating translesion DNA synthesis. We also report that human DNA polymerase eta, a human Y-family DNA polymerase, incorporates the oxidized dNTPs in a similar erroneous manner.

Cationic liposome-mediated gene delivery: biophysical study and mechanism of internalization.
Mohamad Radwan Almofti, Hideyoshi Harashima, Yasuo Shinohara, Ammar Almofti, Yoshinobu Baba, Hiroshi Kiwada
Archives of biochemistry and biophysics, 410, 2, 246, 53, 15 Feb. 2003, [Peer-reviewed], [International Magazine]
English, To identify factors affecting cationic liposome-mediated gene delivery efficiency, we studied the relationship between the biophysical characteristics of liposome/DNA complexes (lipoplexes) at different (+/-) charge ratios, their structures as monitored by atomic force microscopy (AFM), and their mechanism(s) of internalization into the cells. Significant changes were observed in the particle size and zeta potential of liposomes and their structures assessed by AFM upon addition of DNA, which depended on (+/-) charge ratios. AFM images showed that lipoplexes were formed from extensively fused and apparently homogeneous lipid particles encapsulating DNA. Lipoplexes were found to internalize the cells through the endocytosis pathway. Lipoplex-cell fusion was found to occur mainly at the plasma membrane level; however, this lipoplex-cell membrane fusion was found to be essential for the uptake of the large particles. A new perspective for the internalization of large lipoplex particles into cytoplasm is discussed.

Lipoplex size determines lipofection efficiency with or without serum
MR Almofti, H Harashima, Y Shinohara, A Almofti, WH Li, H Kiwada
MOLECULAR MEMBRANE BIOLOGY, 20, 1, 35, 43, Jan. 2003, [Peer-reviewed]
English, Scientific journal

Immunoliposomes having high membrane fluidity delivery a large amount of liposomal doxorubicin into not only doxorubicin sensitive SBC-3 but also doxorubicin resistant SBC-3/ADM cell lines　　　　　　　　　　　　　　　
Kobayashi, T, Okada, Y, Ishida, T, Sone, S, Harashima, H, Maruyama, K, Kiwada, H
J. Liposome Res., 13, 67, 2003, [Peer-reviewed]
English

Utilization efficiency of the oxidized purine nucleotide analogs by DNA polymerase eta.
Naomi Nishimoto, Hiroyuki Kamiya, Hideyoshi Harashima, Shunji Izuta
Nucleic acids research. Supplement (2001), 3, 299, 300, 01 Jan. 2003, [International Magazine]
English, Scientific journal

Mutations induced by 2-hydroxy-dATP during in vitro replication with a HeLa extract.
Kazuya Satou, Hideyoshi Harashima, Hiroyuki Kamiya
Nucleic acids research. Supplement (2001), 325, 326, 01 Jan. 2003

pH-sensitive liposomes in nuclear targeting of macromolecules
R Tachibana, S Futaki, H Harashima, H Kiwada
LIPOSOMES, PT B, 372, 349, 361, 2003, [Peer-reviewed]
English

Size and topology of exogenous DNA as determinant factors of transgene transcription in mammalian cells
H. Kamiya, J. Yamazaki, H. Harashima
Gene Therapy, 9, 1500, 1507, 01 Nov. 2002

Visualization of intracellular trafficking of exogenous DNA delivered by cationic liposomes
H Kamiya, Y Fujimura, Matsuoka, I, H Harashima
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 298, 4, 591, 597, Nov. 2002, [Peer-reviewed]
English, Scientific journal

Quantitative analysis of correlation between number of nuclear plasmids and gene expression activity after transfection with cationic liposomes
R Tachibana, H Harashima, N Ide, S Ukitsu, Y Ohta, N Suzuki, H Kikuchi, Y Shinohara, H Kiwada
PHARMACEUTICAL RESEARCH, 19, 4, 377, 381, Apr. 2002, [Peer-reviewed]
English, Scientific journal

Troglitazone suppresses cell growth of KU812 cells independently of PPAR-gamma
A Abe, Y Kiriyama, M Hirano, T Miura, H Kamiya, H Harashima, Y Tokumitsu
EUROPEAN JOURNAL OF PHARMACOLOGY, 436, 1-2, 7, 13, Feb. 2002
English, Scientific journal

Liposome clearance.
Ishida, T, Harashima, H, Kiwada, H
Biosci. Rep., 22, 197, 224, 2002
English

Pharmacokinetics of targeting with liposomes
H Harashima, T Ishida, H Kamiya, H Kiwada
CRITICAL REVIEWS IN THERAPEUTIC DRUG CARRIER SYSTEMS, 19, 3, 235, 275, 2002
English

Substrate recognition by the human MTH1 protein.
Hiroyuki Kamiya, Laurence Dugué, Hiroyuki Yakushiji, Sylvie Pochet, Yusaku Nakabeppu, Hideyoshi Harashima
Nucleic acids research. Supplement (2001), 2, 85, 6, 2002, [Peer-reviewed], [International Magazine]
English

Intracellular trafficking and transgene expression of viral and non-viral gene vectors
H Kamiya, H Tsuchiya, J Yamazaki, H Harashima
ADVANCED DRUG DELIVERY REVIEWS, 52, 3, 153, 164, Nov. 2001
English

Hydrolysis of oxidized nucleotides by the Escherichia coli Orf135 protein
H Kamiya, N Murata-Kamiya, E Iida, H Harashima
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 288, 3, 499, 502, Nov. 2001
English, Scientific journal

Stearylated arginine-rich peptides: A new class of transfection systems
S. Futaki, W. Ohashi, T. Suzuki, M. Niwa, S. Tanaka, K. Ueda, H. Harashima, Y. Sugiura
Bioconjugate Chemistry, 12, 6, 1005, 1011, Nov. 2001, [Peer-reviewed]
English

Effect of cholesterol content in activation of the classical versus the alternative pathway of rat complement system induced by hydrogenated egg phosphatidylcholine-based liposomes
T Ishida, K Yasukawa, H Kojima, H Harashima, H Kiwada
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 224, 1-2, 69, 79, Aug. 2001, [Peer-reviewed]
English, Scientific journal

The complement system enhances the clearance of phosphatidylserine (PS)-liposomes in rat and guinea pig
TM Huong, T Ishida, H Harashima, H Kiwada
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 215, 1-2, 197, 205, Mar. 2001, [Peer-reviewed]
English, Scientific journal

Interactions of liposomes with in vitro and in vivo: opsonins and receptors.
Ishida, T, Harashima, H, Kiwada, H
Current Drug Metabolism, 2, 397, 409, 2001
English

Molecular design and drug delivery of genetic drugs
Yoshinobu Baba, Mitsuhiko Shionoya, Hideyoshi Harashima
European Journal of Pharmaceutical Sciences, 13, 1, 1, 2001, [Peer-reviewed]
Scientific journal

Quantitative evaluation on relationship between liposome biodistribution in vivo and associated C3 fragments.
Ishida, T, Kojima, H, Harashima, H, Kiwada, H
Int. J. Pharm., 205, No.1-2, 183, 193, 15 Sep. 2000, [Peer-reviewed]
English, Scientific journal, The biodistribution of liposomes with two different kind phospholipids (hydrogenated egg phosphatidylcholine and egg phosphatidylcholine) plus cholesterol (CHOL) were investigated after intravenous administration to rats. Elimination of liposomes from blood circulation was affected by the lipid composition. It appeared that the inclusion of CHOL in liposomes accelerates the rate of liposome uptake by liver, resulting in rapid elimination of liposomes. The amount of C3 fragments bound to liposomes was quantitatively determined to assess the contribution of the complement system to liposome accumulation into organs and liposome destabilization in vivo and in vitro. The amount of bound C3 fragments was directly proportional to CHOL content, and the amount was also proportional to the CLh, CLs as well as CLrel. This relationship suggests that the complement system is responsible for the elimination of liposomes from blood circulation, presumably as a consequence of opsonization by C3 fragments and assembly of membrane attack complex (MAC) onto liposomes. In addition, substitution of cholesteryl methyl ether into the liposome formulation for CHOL significantly diminished not only the binding of C3 fragments but also the CLh, CLs and CLrel, resulting in increased mean resident time (MRT) of the liposomes. This result suggests that the hydroxyl-group on CHOL is a binding site for C3 fragments on the liposomes and that CHOL in a liposome formulation promotes the accumulation of liposomes into the liver and spleen, probably due to their uptake by phagocytic cells, and impairs the stability of the liposomes in blood circulation, via a mechanism involving the complement system.

Biodistribution of liposomes and C3 fragments associated with liposomes: evaluation of their relationship
T Ishida, H Kojima, H Harashima, H Kiwada
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 205, 1-2, 183, 193, Sep. 2000, [Peer-reviewed]
English, Scientific journal

Cardiomegaly in the juvenile visceral steatosis (JVS) mouse is reduced with acute elevation of heart short-chain acyl-carnitine level after L-carnitine injection
Masamichi Kuwajima, Masahisa Horiuchi, Hideyoshi Harashima, Kang-Mo Lu, Miyuki Hayashi, Masako Sei, Kiyokazu Ozaki, Tomoko Kudo, Hiroshi Kamido, Akira Ono, Takeyori Saheki, Kenji Shima
FEBS Letters, 443, 3, 261, 266, 29 Jan. 1999, [Peer-reviewed]
English, Scientific journal

Quantitative assay of transfected plasmid DNA in nucleus using PCR　　　　　　　　　　　　　　　
N. Ide, R. Tachibana, Y. Oota, H. Harashima, Y. Shinohara, H. Kiwada
Proceedings of the Controlled Release Society, 26, 777, 778, 1999
English, International conference proceedings

Kinetic analysis of lipophilic drug encapsulated in liposomes　　　　　　　　　　　　　　　
Y. Ikemoto, K. Masuda, H. Harashima, H. Kiwada
Proceedings of the Controlled Release Society, 26, 757, 758, 1999
English, International conference proceedings

Pharmacokinetics/pharmacodynamic modeling of liposomally encapsulated doxorubicin: Scaling up from rats to humans　　　　　　　　　　　　　　　
H. Doi, H. Harashima, S. Iida, M. Tsuchihashi, H. Kiwada
Proceedings of the Controlled Release Society, 26, 779, 780, 1999
English, International conference proceedings

Incorporation of the nuclear localization signal (NLS) peptide-albumin conjugate to macrophages using pH-sensitive liposomes
S Futaki, R Tachibana, M Shono, M Azumano, M Niwa, H Kiwada, H Harashima
PEPTIDE SCIENCE - PRESENT AND FUTURE, 699, 700, 1999, [Peer-reviewed]
English, International conference proceedings

Intracellular regulation of macromolecules using pH-sensitive liposomes and nuclear localization signal: Qualitative and quantitative evaluation of intracellular trafficking
R Tachibana, H Harashima, M Shono, M Azumano, M Niwa, S Futaki, H Kiwada
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 251, 2, 538, 544, Oct. 1998, [Peer-reviewed]
English, Scientific journal

Complement dependent and independent liposome uptake by peritoneal macrophages: Cholesterol content dependency
TM Huong, H Harashima, H Kiwada
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 21, 9, 969, 973, Sep. 1998, [Peer-reviewed]
English, Scientific journal

Kinetic analysis of the interaction between liposomes and the complement system in rat serum: Re-evaluation of size-dependency
M Yamada, H Harashima, H Kiwada
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 21, 9, 964, 968, Sep. 1998, [Peer-reviewed]
English, Scientific journal

Mechanism of initial distribution of blood-borne colon carcinoma cells in the liver
N Mizuno, Y Kato, K Shirota, Y Izumi, T Irimura, H Harashima, H Kiwada, N Motoji, A Shigematsu, Y Sugiyama
JOURNAL OF HEPATOLOGY, 28, 5, 878, 885, May 1998, [Peer-reviewed]
English, Scientific journal

Effects of repeated clarithromycin administration on the pharmacokinetic properties of pindolol in rats
Takafumi Komori, Kazuki Shimoishi, Hideyoshi Harashima, Masaki Otagiri
Biological and Pharmaceutical Bulletin, 21, 12, 1376, 1378, Pharmaceutical Society of Japan, 1998, [Peer-reviewed]
English, Scientific journal

Computer simulation of the effects of alterations in blood flows and body composition on thiopental pharmacokinetics in humans
D. Russell Wada, Sven Björkman, William F. Ebling, Hideyoshi Harashima, Sandra R. Harapat, Donald R. Stanski
Anesthesiology, 87, 4, 884, 899, Oct. 1997, [Peer-reviewed]
English, Scientific journal

No effect of age on the dose requirement of thiopental in the rat
Hideyoshi Harashima, William F. Ebling, D. Russell Wada, Donald R. Stanski
Experimental Gerontology, 32, 3, 315, 324, May 1997, [Peer-reviewed]
English, Scientific journal

Kinetic analysis of the interaction between liposomes and complement system in rat serum　　　　　　　　　　　　　　　
M. Yamada, H. Harashima, J. Kikuishi, Y. Nemoto, M. Shibuya, H. Kiwada
Proceedings of the Controlled Release Society, 24, 793, 794, 1997
English, International conference proceedings

Verification of satiated model in the saturable hepatic uptake clearance of liposomes　　　　　　　　　　　　　　　
T. Shimada, H. Harashima, H. Kiwada
Proceedings of the Controlled Release Society, 24, 569, 570, 1997
English, International conference proceedings

Synergistic effect between size and cholesterol content in the enhanced hepatic uptake clearance of liposomes through complement activation in rats
H Harashima, TM Huong, T Ishida, Y Manabe, H Matsuo, H Kiwada
PHARMACEUTICAL RESEARCH, 13, 11, 1704, 1709, Nov. 1996, [Peer-reviewed]
English, Scientific journal

Species difference in the disposition of liposomes among mice, rats, and rabbits: Allometric relationship and species dependent hepatic uptake mechanism
H Harashima, S Komatsu, S Kojima, C Yanagi, Y Morioka, M Naito, H Kiwada
PHARMACEUTICAL RESEARCH, 13, 7, 1049, 1054, Jul. 1996, [Peer-reviewed]
English, Scientific journal

Carnitine transport defect in fibroblasts of juvenile visceral steatosis (JVS) mouse
Masamichi Kuwajima, Kang-Mo Lu, Hideyoshi Harashima, Akira Ono, Izumi Sato, Akira Mizuno, Takashi Murakami, Hiromu Nakajima, Jun-Ichiro Miyagawa, Mitsuyoshi Namba, Toshiaki Hanafusa, Jun-Ichiro Hayakawa, Yuji Matsuzawa, Kenji Shima
Biochemical and Biophysical Research Communications, 223, 2, 283, 287, Academic Press Inc., 14 Jun. 1996, [Peer-reviewed]
English, Scientific journal

Effects of thiopental on regional blood flows in the rat
D. Russell Wada, Hideyoshi Harashima, William F. Ebling, Eileen W. Osaki, Donald R. Stanski
Anesthesiology, 84, 3, 596, 604, Mar. 1996, [Peer-reviewed]
English, Scientific journal

Size-dependent release of carboxyfluorescein from cetylmannoside-modified liposomes in human plasma
AJ Ferdous, T Ishida, M Shinohara, H Harashima, H Kiwada
BIOPHARMACEUTICS & DRUG DISPOSITION, 17, 2, 145, 154, Mar. 1996
English, Scientific journal

Studies on the mechanism of uptake of liposomes using an isolated perfused liver system
Hideyoshi Harashima, Hiroshi Kiwada
Journal of Liposome Research, 6, 1, 61, 75, Informa Healthcare, 1996, [Peer-reviewed]
English, Scientific journal

Simulation study on the optimization of antitumor effect of liposomally encapsulated doxorubicin based on the physiological model with cell kill kinetic model
S Iida, Y Urakami, H Harashima, H Kiwada
23RD INTERNATIONAL SYMPOSIUM ON CONTROLLED RELEASE OF BIOACTIVE MATERIALS, 1996 PROCEEDINGS, 653, 654, 1996, [Peer-reviewed]
English, International conference proceedings

The effect of dose on the intracellular fate of liposomes
K Yoshimura, Y Morioka, H Harashima, H Kiwada
23RD INTERNATIONAL SYMPOSIUM ON CONTROLLED RELEASE OF BIOACTIVE MATERIALS, 1996 PROCEEDINGS, 683, 684, 1996, [Peer-reviewed]
English, International conference proceedings

Effect between size and cholesterol content in the enhanced hepatic uptake clearance of liposomes through complement activation in rats　　　　　　　　　　　　　　　
Harashima, H, Huong, T.M, Ishida, T, Manabe, Y, Matsuo, H, Kiwada, H
Progress in Drug Delivery System, V, 93, 96, 1996
English

EFFECTS OF THE SIZE AND FLUIDITY OF LIPOSOMES ON THEIR ACCUMULATION IN TUMORS - A PRESUMPTION OF THEIR INTERACTION WITH TUMORS
K UCHIYAMA, A NAGAYASU, Y YAMAGIWA, T NISHIDA, H HARASHIMA, H KIWADA
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 121, 2, 195, 203, Jul. 1995, [Peer-reviewed]
English, Scientific journal

KINETIC MODELING OF LIPOSOME DEGRADATION IN PERITONEAL-MACROPHAGES
H HARASHIMA, N HIRAI, H KIWADA
BIOPHARMACEUTICS & DRUG DISPOSITION, 16, 2, 113, 123, Mar. 1995, [Peer-reviewed]
English, Scientific journal

Size dependent liposome degradation in blood: In vivo/in vitro correlation by kinetic modeling
H Harashima, T Hiraiwa, Y Ochi, H Kiwada
JOURNAL OF DRUG TARGETING, 3, 4, 253, 261, 1995, [Peer-reviewed]
English, Scientific journal

Size dependent characterization of cetylmannoside-modified liposomes (Man-MLVs) in human plasma　　　　　　　　　　　　　　　
Ishida, T, Ferdous, A.J, Shinohara, M, Harashima, H, Kiwada, H
J.Liposome Res., 6, 252, 1995
English

KINETIC MODELING OF LIPOSOME DEGRADATION IN SERUM - EFFECT OF SIZE AND CONCENTRATION OF LIPOSOMES IN-VITRO
H HARASHIMA, Y OCHI, H KIWADA
BIOPHARMACEUTICS & DRUG DISPOSITION, 15, 3, 217, 225, Apr. 1994, [Peer-reviewed]
English, Scientific journal

ENHANCED HEPATIC-UPTAKE OF LIPOSOMES THROUGH COMPLEMENT ACTIVATION DEPENDING ON THE SIZE OF LIPOSOMES
H HARASHIMA, K SAKATA, K FUNATO, H KIWADA
PHARMACEUTICAL RESEARCH, 11, 3, 402, 406, Mar. 1994, [Peer-reviewed]
English, Scientific journal

COMPLEMENT-MEDIATED HEPATIC UPTAKE OF MANNOSIDE-MODIFIED LIPOSOMES
MATSUO Hirotami, FUNATO Koichi, YAMASHITA Chikamasa, HARASHIMA Hideyoshi, KIWADA Hiroshi
Drug Metabolism and Pharmacokinetics, 9, 118, 121, The Japanese Society for the Study of Xenobiotics, 1994
English, In the elimination of injected liposomes in vivo, it is considered that several serum components play an important role on hepatic uptake of them. This study was conducted to clarify the hepatic uptake mechanism of cetylmannoside (Man) -modified multilamellar vesicles (Man-MLV) using perfused rat liver. In the presence of serum, Man-MLV was taken up by the liver depending on the serum concentration, and it showed an approximately 2-fold higher accumulation than non-modified MLV (PC-MLV). These hepatic uptake of liposomes were obviously inhibited by preheating the serum at 56 °C for 30 min or by the treatment with anti-rat C3 antiserum. Further, SDS-PAGE followed by immunoblot analysis showed the deposition of iC3b on the opsonized Man-MLV. These results obtained in the present study suggested that hepatic uptake of Man-MLV was mainly mediated by complement receptor rather than mannose receptor on Kupffer cells in vivo.

THE COMPLEMENT-MEDIATED BUT NOT MANNOSE RECEPTOR-MEDIATED PHAGOCYTOSIS IS INVOLVED IN THE HEPATIC-UPTAKE OF CETYLMANNOSIDE-MODIFIED LIPOSOMES IN-SITU
H MATSUO, K FUNATO, H HARASHIMA, H KIWADA
JOURNAL OF DRUG TARGETING, 2, 2, 141, 146, 1994, [Peer-reviewed]
English, Scientific journal

KINETIC-ANALYSIS OF TISSUE DISTRIBUTION OF DOXORUBICIN INCORPORATED IN LIPOSOMES IN RATS .2.
H HARASHIMA, Y MIDORI, S OHSHIMA, K YACHI, H KIKUCHI, H KIWADA
BIOPHARMACEUTICS & DRUG DISPOSITION, 14, 7, 595, 608, Oct. 1993, [Peer-reviewed]
English, Scientific journal

Tissue distribution of fentanyl and alfentanil in the rat cannot be described by a blood flow limited model
Sven Björkman, Donald R. Stanski, Hideyoshi Harashima, Robert Dowrie, Sandra R. Harapat, D.Russell Wada, William F. Ebling
Journal of Pharmacokinetics and Biopharmaceutics, 21, 3, 255, 279, Kluwer Academic Publishers-Plenum Publishers, Jun. 1993, [Peer-reviewed]
English, Scientific journal

KINETIC-ANALYSIS OF AUC-DEPENDENT SATURABLE CLEARANCE OF LIPOSOMES - MATHEMATICAL-DESCRIPTION OF AUC DEPENDENCY
H HARASHIMA, C YAMANE, Y KUME, H KIWADA
JOURNAL OF PHARMACOKINETICS AND BIOPHARMACEUTICS, 21, 3, 299, 308, Jun. 1993, [Peer-reviewed]
English, Scientific journal

KINETIC MODELING OF LIPOSOME DEGRADATION IN BLOOD-CIRCULATION
H HARASHIMA, Y KUME, C YAMANE, H KIWADA
BIOPHARMACEUTICS & DRUG DISPOSITION, 14, 3, 265, 270, Apr. 1993, [Peer-reviewed]
English, Scientific journal

DISTINCTION BETWEEN THE DEPLETION OF OPSONINS AND THE SATURATION OF UPTAKE IN THE DOSE-DEPENDENT HEPATIC-UPTAKE OF LIPOSOMES
H HARASHIMA, K SAKATA, H KIWADA
PHARMACEUTICAL RESEARCH, 10, 4, 606, 610, Apr. 1993, [Peer-reviewed]
English, Scientific journal

KINETIC ANALYSIS OF LIPOSOME UPTAKE BY RES IN COMPARISON TO THEIR DEGRADATION IN BLOOD AND RES
Harashima H., Ochi Y., Hiraiwa T., Sugino K., Kiwada H.
Drug Metabolism and Pharmacokinetics, 8, 751, 754, The Japanese Society for the Study of Xenobiotics, 1993
English, Kinetic analysis of the hepatic uptake of liposomes was performed in comparison to the degradation of liposomes in blood and liver. The degradation clearance of liposomes in blood was shown to depend on the size of liposomes. In vitro kinetic study on the degradation of liposomes suggested the mechanism of this size dependency due to the Increased affinity of liposomes for complement. There was two kinds of hepatic uptake components of liposomes, one is a size dependent opsonized pathway, and the other is a size Independent unopsonized pathway. The size dependent opsonization of liposomes was shown to result from the size dependent complement activation and this component explained the size dependended hepatic uptake clearance of liposomes in vivo. The kinetic analysis of the hepatic degradation of liposomes was also performed by labeling liposomes with both ¹²⁵I-albumin and ³H-cholesterylhexadecylether. Although the mechanism of hepatic uptake are considered to be different depending on the size of liposomes, the kinetics of hepatic degradation of liposomes was same for each size of liposomes and lay in the order of hour.

KINETIC-ANALYSIS OF TISSUE DISTRIBUTION OF DOXORUBICIN INCORPORATED IN LIPOSOMES IN RATS .1.
H HARASHIMA, S OHSHIMA, Y MIDORI, K YACHI, H KIKUCHI, H KIWADA
BIOPHARMACEUTICS & DRUG DISPOSITION, 13, 3, 155, 170, Apr. 1992, [Peer-reviewed]
English, Scientific journal

Kinetic Modeling of Ouabain Tissue Distribution Based on Slow and Saturable Binding to Na,K-ATPase
Hideyoshi Harashima, Michio Mamiya, Masayo Yamazaki, Yasufumi Sawada, Tatsuji Iga, Manabu Hanano, Yuichi Sugiyama
Pharmaceutical Research: An Official Journal of the American Association of Pharmaceutical Scientists, 9, 12, 1607, 1611, 1992, [Peer-reviewed]
English, Scientific journal

Significance of Binding to Na,K-ATPase in the Tissue Distribution of Ouabain in Guinea Pigs
Hideyoshi Harashima, Michio Mamiya, Masayo Yamazaki, Yuichi Sugiyama, Yasufumi Sawada, Tatsuji Iga, Manabu Hanano
Pharmaceutical Research: An Official Journal of the American Association of Pharmaceutical Scientists, 9, 4, 474, 479, 1992, [Peer-reviewed]
English, Scientific journal

SATURABLE, NON-MICHAELIS-MENTEN UPTAKE OF LIPOSOMES BY THE RETICULOENDOTHELIAL SYSTEM
Y KUME, F MAEDA, H HARASHIMA, H KIWADA
JOURNAL OF PHARMACY AND PHARMACOLOGY, 43, 3, 162, 166, Mar. 1991, [Peer-reviewed]
English, Scientific journal

Kinetic analysis of the dose-dependent hepatic handling of 1-anilino-8-naphthalene sulfonate in rats
Youn Bok Chung, Seiji Miyauchi, Yuichi Sugiyama, Hideyoshi Harashima, Tatsuji Iga, Manabu Hanano
Journal of Pharmacokinetics and Biopharmaceutics, 18, 4, 313, 333, Kluwer Academic Publishers-Plenum Publishers, Aug. 1990, [Peer-reviewed]
English, Scientific journal

Effect of various organic anions on the plasma disappearance of 1-anilino-8-naphthalene sulfonate
Youn Bok Chung, Seiji Miyauchi, Yuichi Sugiyama, Hideyoshi Harashima, Tatsuji Iga, Manabu Hanano
Journal of Hepatology, 11, 2, 240, 251, 1990, [Peer-reviewed]
English, Scientific journal

NONLINEAR TISSUE DISTRIBUTION OF OUABAIN IN RABBITS
H HARASHIMA, Y SUGIYAMA, T IGA, M HANANO
DRUG METABOLISM AND DISPOSITION, 16, 4, 645, 649, Jul. 1988, [Peer-reviewed]
English, Scientific journal

INFUSION RATE-DEPENDENT POSITIVE INOTROPIC ACTION OUABAIN IN RABBITS
H HARASHIMA, Y SUGIYAMA, Y SAWADA, K SHIGENOBU, Y KASUYA, T IGA, M HANANO
CHEMICAL & PHARMACEUTICAL BULLETIN, 35, 7, 2923, 2927, Jul. 1987, [Peer-reviewed]
English, Scientific journal

ANALYSIS OF NONLINEAR TISSUE DISTRIBUTION OF QUINIDINE IN RATS BY PHYSIOLOGICALLY BASED PHARMACOKINETICS
H HARASHIMA, Y SAWADA, Y SUGIYAMA, T IGA, M HANANO
JOURNAL OF PHARMACOKINETICS AND BIOPHARMACEUTICS, 13, 4, 425, 440, Aug. 1985, [Peer-reviewed]
English, Scientific journal

PREDICTION OF THE VOLUMES OF DISTRIBUTION OF BASIC DRUGS IN HUMANS BASED ON DATA FROM ANIMALS
Y SAWADA, M HANANO, Y SUGIYAMA, H HARASHIMA, T IGA
JOURNAL OF PHARMACOKINETICS AND BIOPHARMACEUTICS, 12, 6, 587, 596, 1984, [Peer-reviewed]
English, Scientific journal

ポリカチオン/DNA複合体搭載脂質ナノ粒子による効率的な長鎖DNAトランスフェクション
宇野秀哉, 真栄城正寿, 真栄城正寿, 真栄城正寿, 佐藤悠介, 石田晃彦, 日比野光恵, 谷博文, 原島秀吉, 渡慶次学, 化学系学協会北海道支部冬季研究発表会(Web), 2023, 2023

抗酸化能を有した心筋再生治療用ミトコンドリア活性化ヒト由来心筋前駆細胞の開発
白石真大, 白石真大, 佐々木大輔, 日比野光恵, 日比野光恵, 武田充人, 原島秀吉, 山田勇磨, 山田勇磨, 日本酸化ストレス学会学術集会プログラム・抄録集, 76th, 2023

Effect of Lipid Nanoparticle Properties on the Therapeutic Effects against Acetaminophen Liver Injury
日比野光恵, 日比野光恵, 真栄城正寿, 渡慶次学, 原島秀吉, 山田勇磨, 日本薬剤学会年会講演要旨集(CD-ROM), 38th, 2023

難水溶性物質CoQ₁₀を搭載可能な脂質ナノ粒子設計の検討
杉原崚太, 日比野光恵, 真栄城正寿, 渡慶次学, 中村孝司, 原島秀吉, 山田勇磨, 生体膜と薬物の相互作用シンポジウム講演要旨集, 44th, 2023

Validation of mitochondrial-derived component supplementation strategy for the purpose of activating mitochondrial function
伊藤百, 山田勇磨, 山田勇磨, 真栄城正寿, 渡慶次学, 原島秀吉, 日本薬学会年会要旨集(Web), 143rd, 2023

Evaluation of an innovative mitochondrial genome-editing system using mitochondria-targeted nanoparticles in vitro
野呂田楓, 石塚宣, 廣瀬みさ, 真栄城正寿, 渡慶次学, 原島秀吉, 山田勇磨, 山田勇磨, 日本薬学会年会要旨集(Web), 143rd, 2023

特性の異なるコエンザイムQ₁₀搭載脂質ナノ粒子を用いたアセトアミノフェン肝障害に対する治療効果の検証
日比野光恵, 真栄城正寿, 渡慶次学, 原島秀吉, 山田勇磨, 日本DDS学会学術集会プログラム予稿集, 39th, 2023

CRISPR/Cas9システムを用いたmtDNAゲノム編集の検証
野呂田楓, 野呂田楓, 野呂田楓, 石塚宣, 廣瀬みさ, 真栄城正寿, 渡慶次学, 原島秀吉, 山田勇磨, 日本DDS学会学術集会プログラム予稿集, 39th, 2023

癌光治療用ミトコンドリア標的型ナノカプセルの殺細胞効果向上を目指した試み
長縄莉奈, 高野勇太, 高野勇太, 趙韓俊, 真栄城正寿, 渡慶次学, 原島秀吉, 山田勇磨, 日本DDS学会学術集会プログラム予稿集, 39th, 2023

薬剤送達ナノ粒子作製用ガラス製マイクロ流体システムの開発
岡田悠斗, 真栄城正寿, 真栄城正寿, 佐藤悠介, 石田晃彦, 谷博文, 原島秀吉, 渡慶次学, 化学系学協会北海道支部冬季研究発表会(Web), 2022, 2022

ミトコンドリア標的型DDSを基盤とした癌光治療の検証
山田勇磨, 窪田文佳, 高野勇太, BIJU Vasudevanpillai, 真栄城正寿, 渡慶次学, 繁富香織, 宮武由甲子, 原島秀吉, 日本DDS学会学術集会プログラム予稿集, 38th, 2022

コエンザイムQ₁₀搭載ミトコンドリア標的型ナノカプセル注射剤の開発
日比野光恵, 山田勇磨, 真栄城正寿, 渡慶次学, 原島秀吉, 生体膜と薬物の相互作用シンポジウム講演要旨集, 43rd, 2022

心筋再生治療用ミトコンドリア活性化ヒト由来心筋幹細胞の開発
白石真大, 白石真大, 山田勇磨, 山田勇磨, 佐々木大輔, 日比野光恵, 武田充人, 原島秀吉, 遺伝子・デリバリー研究会シンポジウム要旨集, 21st (CD-ROM), 2022

蛍光ナノダイヤモンド搭載セラノスティクス用脂質ナノ粒子の作製
杉浦魁星, 真栄城正寿, 真栄城正寿, 佐藤悠介, 石田晃彦, 谷博文, 原島秀吉, 渡慶次学, 化学とマイクロ・ナノシステム学会研究会講演要旨集(CD-ROM), 45th, 2022

マイクロ流体デバイスを用いた長鎖プラスミドDNA搭載ナノ粒子の作製
宇野秀哉, 真栄城正寿, 真栄城正寿, 佐藤悠介, 石田晃彦, 原島秀吉, 渡慶次学, 化学とマイクロ・ナノシステム学会研究会講演要旨集(CD-ROM), 46th, 2022

Construction of preparation method using microfluidic device and validation of therapeutic effect for mitochondrial targeted coenzyme Q₁₀ encapsulated nanocapsule injectable drug
日比野光恵, 山田勇磨, 真栄城正寿, 渡慶次学, 原島秀吉, 日本薬学会年会要旨集(Web), 142nd, 2022

Validation of mitochondrial component supplementation strategy to activate mitochondrial function
伊藤百, 山田勇磨, 山田勇磨, 日比野光恵, 佐々木大輔, 真栄城正寿, 渡慶次学, 渡慶次学, 太田善浩, 原島秀吉, 原島秀吉, 日本薬学会年会要旨集(Web), 142nd, 2022

ミトコンドリア活性化ヒト由来心筋幹細胞を用いた細胞移植療法の検証
白石真大, 白石真大, 山田勇磨, 山田勇磨, 佐々木大輔, 日比野光恵, 真栄城正寿, 渡慶次学, 武田充人, 原島秀吉, 日本DDS学会学術集会プログラム予稿集, 38th, 2022

Establishment of Preparation Method for Lipid Nanoparticles Encapsulating Poorly Water-Soluble Drug by Using Microfluidic Device
日比野光恵, 日比野光恵, 山田勇磨, 山田勇磨, 原島秀吉, 製剤機械技術学会誌, 31, 4, 2022

【モダリティ多様化を支援する創薬基盤技術】創薬におけるパラダイムシフトと大学発革新的ナノ医療の創出
原島 秀吉, ファルマシア, 57, 7, 630, 634, Jul. 2021, [Invited], [Corresponding author]
(公社)日本薬学会, Japanese, Introduction scientific journal

ミトコンドリア標的型光線力学的療法を可能とするナノキャリアの構築及び機能評価
窪田文佳, 山田勇磨, SATRIALDI, 高野勇太, 真栄城正寿, 渡慶次学, 原島秀吉, 生体膜と薬物の相互作用シンポジウム講演要旨集, 42nd (CD-ROM), 2021

Evaluation of therapeutic effect of myocardial stem cell transplantation using human mitochondria-enhanced cardiac stem cells in a rat model of acute heart failure
佐々木大輔, 佐々木大輔, 山田勇磨, 山田勇磨, 後藤悠太, 白石真大, 白石真大, 武田充人, 原島秀吉, 原島秀吉, 日本薬剤学会年会講演要旨集(CD-ROM), 36th, 2021

心筋ミトコンドリア標的型リポソームを用いたドキソルビシン心筋傷害軽減の試み
辻岡孝郎, 辻岡孝郎, 山田勇磨, 佐々木大輔, 武田充人, 原島秀吉, 日本腫瘍循環器学会学術集会抄録集(Web), 4th, 2021

マイクロ流体デバイスを用いたプラスミドDNA搭載脂質ナノ粒子の作製
宇野秀哉, 真栄城正寿, 真栄城正寿, 佐藤悠介, 石田晃彦, 谷博文, 原島秀吉, 渡慶次学, 化学とマイクロ・ナノシステム学会研究会講演要旨集, 43rd, 2021

炎症性腸疾患におけるグルコシルセラミドの機能解析
小室茉莉子, 永根大幹, 丹羽智瑛, 宮本貴祥, 遠藤力斗, 中村孝司, 原島秀吉, 相原尚之, 上家潤一, 山下匡, 日本酸化ストレス学会学術集会プログラム・抄録集, 74th (CD-ROM), 2021

炎症性腸疾患におけるグルコシルセラミドの機能解析
小室茉莉子, 永根大幹, 宮本貴祥, 丹羽智瑛, 遠藤力斗, 中村孝司, 原島秀吉, 相原尚之, 上家潤一, 山下匡, 日本生化学会大会(Web), 94th, 2021

Trans MIT system
山田勇磨, 山田勇磨, 日比野光恵, 伊藤百, 荒井愛永, 佐々木大輔, 真栄城正寿, 渡慶次学, 渡慶次学, 太田善浩, 原島秀吉, 原島秀吉, 日本薬剤学会年会講演要旨集(CD-ROM), 36th, 2021

Properties of lipid nanoparticles affecting their lymph node transitivity and distribution
中村孝司, 河合美典, 佐藤悠介, 真栄城正寿, 渡慶次学, 原島秀吉, 日本薬剤学会年会講演要旨集(CD-ROM), 36th, 2021

多機能性エンベロープ型ナノ構造体の開発とナノ医療への展開　　　　　　　　　　　　　　　
原島 秀吉, 日本小児臨床薬理学会雑誌, 33, 1, 88, 88, Dec. 2020, [Invited], [Corresponding author]
日本小児臨床薬理学会, Japanese

ミトコンドリアを標的とする遺伝子治療用RNAナノカプセルの創製　　　　　　　　　　　　　　　
山田 勇磨, 原島 秀吉, 遺伝子医学, 10, 4, 74, 79, Oct. 2020, [Invited], [Last author]
(株)メディカルドゥ, Japanese

ミトコンドリア強化幹細胞(MITO Cell)の製造および細胞移植療法への展開　　　　　　　　　　　　　　　
山田 勇磨, 阿部 二郎, 佐々木 大輔, 原島 秀吉, BIO Clinica, 35, 5, 448, 450, May 2020, [Invited], [Last author]
(株)北隆館, Japanese

【核酸医薬の最近の話題】核酸DDS技術を用いたがん免疫療法　　　　　　　　　　　　　　　
中村 孝司, 原島 秀吉, 細胞, 52, 5, 245, 248, May 2020, [Invited], [Last author]
(株)ニュー・サイエンス社, Japanese

【核酸創薬に貢献するバイオマテリアル】バイオマテリアルに基づいた核酸ナノ医療の創製　　　　　　　　　　　　　　　
山田 勇磨, 中村 孝司, 佐藤 悠介, 原島 秀吉, バイオマテリアル-生体材料-, 38, 2, 92, 99, Apr. 2020, [Invited], [Last author]
日本バイオマテリアル学会, Japanese

T細胞のグルコシルセラミドは炎症性大腸炎を抑制する
永根大幹, 山門みのり, 水野愛理, 遠藤力斗, 中村孝司, 原島秀吉, 相原尚之, 上家潤一, 山下匡, 日本酸化ストレス学会学術集会プログラム・抄録集, 73rd, 2020

Size and charge effect of lipid nanoparticles on transitivity to lymph node and distribution in lymph node
中村孝司, 河合美典, 佐藤悠介, 真栄城正寿, 渡慶次学, 原島秀吉, 日本薬学会年会要旨集(CD-ROM), 140th (Web), 2020

CRISPR/Casタンパク質搭載脂質ナノ粒子の開発
佐藤悠介, 鈴木裕一, 小沼はるの, 佐藤里咲, 真栄城正寿, 渡慶次学, 原島秀吉, 日本DDS学会学術集会プログラム予稿集, 36th, 2020

コエンザイムQ10封入ミトコンドリア標的型DDSの開発およびアセトアミノフェン肝障害モデルへの治療効果の検証
日比野光恵, 山田勇磨, 真栄城正寿, 渡慶次学, 石塚洋一, 原島秀吉, 日本DDS学会学術集会プログラム予稿集, 36th, 2020

ガラス製マイクロ流体デバイスを用いたsiRNA搭載脂質ナノ粒子の作製と大量生産用集積化デバイスの開発
真栄城正寿, 真栄城正寿, 岡田悠斗, 佐藤悠介, 石田晃彦, 谷博文, 原島秀吉, 渡慶次学, 日本DDS学会学術集会プログラム予稿集, 36th, 2020

DDSナノ粒子作製用ガラス製マイクロ流体デバイスの開発
岡田悠斗, 真栄城正寿, 真栄城正寿, 佐藤悠介, 石田晃彦, 谷博文, 原島秀吉, 渡慶次学, 化学とマイクロ・ナノシステム学会研究会講演要旨集, 42nd (Web), 2020

平成維新-平成30年を振り返る- 平成のDDS(siRNA/リポソーム編)を原島先生と共に振り返る MEND(細胞内制御)とPEGジレンマはこうして誕生した
原島 秀吉, 畠山 浩人, 金沢 貴憲, 薬剤学: 生命とくすり, 80, 1, 13, 23, Jan. 2020, [Invited], [Lead author]
(公社)日本薬剤学会, Japanese

Recent advances in the targeting of systemically administered non-viral gene delivery systems
Ikramy A. Khalil, Yusuke Sato, Hideyoshi Harashima, Expert Opinion on Drug Delivery, 16, 10, 1037, 1050, 03 Oct. 2019
Taylor and Francis Ltd, English, Book review

【ミトコンドリアと疾患・老化 細胞内代謝プラントとしての役割を知り、ミトコンドリアを標的とした創薬に挑む】(第3章)ミトコンドリア疾患の診断技術と治療戦略 治療技術・治療薬開発 ミトコンドリア標的型ナノDDSが創る未来医療
山田 勇磨, 原島 秀吉, 実験医学, 37, 12, 2067, 2073, Aug. 2019, [Invited], [Last author]
(株)羊土社, Japanese

心不全病態に対する多面的アプローチ 基礎から治療へ 虚血再灌流モデルマウスを用いた心不全に対するミトコンドリア活性化心筋幹細胞を用いた細胞移植療法の治療効果の検討　　　　　　　　　　　　　　　
佐々木 大輔, 阿部 二郎, 武田 充人, 原島 秀吉, 山田 勇磨, 日本小児循環器学会雑誌, 35, Suppl.1, s1, 125, Jun. 2019
(NPO)日本小児循環器学会, Japanese

【小児科医に知ってほしいミトコンドリア病UPDATE】治療の展望 ミトコンドリアDDSが拓く新しいミトコンドリア病治療戦略
山田 勇磨, 原島 秀吉, 小児科診療, 82, 4, 523, 528, Apr. 2019, [Invited], [Last author]
(株)診断と治療社, Japanese

【加速する核酸医薬開発】siRNAのDDS開発の現状と展望
佐藤 悠介, 原島 秀吉, 医薬ジャーナル, 55, 2, 615, 619, Feb. 2019, [Invited], [Last author]
(株)医薬ジャーナル社, Japanese

実験計画法によるmRNA送達キャリアの効率的な最適化
橋場月, 佐藤悠介, 真栄城正寿, 渡慶次学, 原島秀吉, 日本核酸医薬学会年会講演要旨集, 5th, 2019

ゲノム編集タンパク質搭載脂質ナノ粒子の開発
佐藤悠介, 鈴木裕一, 佐藤里咲, 真栄城正寿, 渡慶次学, 原島秀吉, 日本核酸医薬学会年会講演要旨集, 5th, 2019

脂質ナノ粒子形成後の二次希釈操作が粒子サイズへ与える影響の解明
木村笑, 真栄城正寿, 岡部奈々, 佐藤悠介, 石田晃彦, 谷博文, 原島秀吉, 渡慶次学, 化学とマイクロ・ナノシステム学会研究会講演要旨集, 39th, 2019

ミトコンドリアへの分子送達技術　　　　　　　　　　　　　　　
山田 勇磨, 原島 秀吉, BIO Clinica, 33, 7, 637, 640, Jul. 2018, [Invited], [Last author]
(株)北隆館, Japanese

【"EPR効果"】EPR効果と血管を標的としたがん微小環境制御によるナノ粒子送達戦略
櫻井 遊, 原島 秀吉, Drug Delivery System, 33, 2, 98, 104, Mar. 2018, [Invited], [Last author, Corresponding author]
日本DDS学会, Japanese

ミトコンドリア強化幹細胞(MITO Cell)の製造および心筋症モデルを用いた細胞移植療法の検証
山田勇磨, 阿部二郎, 佐々木大輔, 武田充人, 原島秀吉, 日本酸化ストレス学会学術集会プログラム・抄録集, 71st, 2018

ミトコンドリア活性化幹細胞を用いた心筋幹細胞移植療法の確立
佐々木大輔, 山田勇磨, 阿部二郎, 武田充人, 原島秀吉, 次世代を担う若手医療薬科学シンポジウム抄録集, 12th, 2018

iLiNPデバイスによるpH応答性カチオン性脂質ナノ粒子の粒径制御
木村笑, 真栄城正寿, 岡部奈々, 佐藤悠介, 石田晃彦, 谷博文, 原島秀吉, 渡慶次学, 渡慶次学, 渡慶次学, 渡慶次学, 化学とマイクロ・ナノシステム学会研究会講演要旨集, 37th, 2018

マイクロデバイスを用いた脂質ナノ粒子作製と生体内動態挙動
真栄城正寿, 木村笑, 佐藤悠介, 石田晃彦, 谷博文, 原島秀吉, 渡慶次学, 化学工学会秋季大会研究発表講演要旨集(CD-ROM), 49th, 2017

Strategy for multiple cancer immunotherapy based on c-di-GMP loaded lipid nanoparticles
中村 孝司, 原島 秀吉, 細胞, 48, 13, 655, 659, Dec. 2016
ニューサイエンス社, Japanese

Development of pH-Sensitive Lipid -Based Lipid Nanoparticles and Its Application to Nucleic Acid Nanomedicine
佐藤 悠介, 原島 秀吉, 化學工業, 67, 11, 821, 827, Nov. 2016
小峰工業出版, Japanese

pH感受性脂質を基盤とした脂質ナノ粒子の開発と核酸ナノメディシンへの応用　　　　　　　　　　　　　　　
佐藤悠介, 原島秀吉, 化学工業, 67, 11, 21, 27, Nov. 2016, [Invited], [Last author, Corresponding author]

【DDSを支える高分子-ポリエチレングリコール-】PEGジレンマ 生体内安定性と細胞内動態の制御によるがんへの遺伝子・核酸デリバリー
畠山 浩人, 原島 秀吉, Drug Delivery System, 31, 4, 293, 299, Sep. 2016, [Invited], [Last author, Corresponding author]
日本DDS学会, Japanese

A Lipid Nanoparticle for Efficient Delivery of siRNA to Dendritic Cells Towards Cell-based Cancer Therapy
中村 孝司, 原島 秀吉, 化學工業, 67, 7, 477, 482, Jul. 2016
小峰工業出版, Japanese

Formation of Lipid Nanoparticles Using Microfluidic Devices and its Application for DDS
真栄城正寿, 佐藤悠介, 原島秀吉, 渡慶次学, 機能材料, 36, 7, 15, 21, Jul. 2016, [Invited]
シーエムシー出版, Japanese, Introduction commerce magazine

Properties and Evaluation of Liposome Drug Product
加藤 くみ子, 石原 比呂之, 友田 寬, 松田 嘉弘, 永井 尚美, 花田 博幸, 久田 茂, 小野寺 博志, 西山 伸宏, 原島 秀吉, 松村 保広, 片岡 一則, 合田 幸広, 奥田 晴宏, 川西 徹, 医薬品医療機器レギュラトリーサイエンス = Pharmaceutical and medical device regulatory science, 47, 5, 333, 341, May 2016
リポソーム製剤は有効成分をリポソームの脂質二分子膜または内相に封入することにより作製される。今回、ナノメートルサイズのリポソーム製剤で、有効成分の生体内安定性、組織移行性プロファイルをはじめとする薬物動態、細胞内動態等の向上を目的として設計された製剤について、以下の項目に分けて述べた。1)リポソーム製剤に特有の品質特性と製法変更時の同等性/同質性評価、2)非臨床薬物動態、3)非臨床毒性、として述べた。, 日本公定書協会, Japanese

プラスミドDNA結合ヒストンの特異的アセチル化による外来遺伝子発現上昇
紙谷浩之, 紙谷浩之, 紙谷浩之, 西原実香, 西原実香, 神田元紀, 神田元紀, 鈴木哲矢, 山門振一郎, 原島秀吉, 日本DDS学会学術集会プログラム予稿集, 32nd, 2016

血管を標的とする薬物送達システムに基づいた新しい治療法　　　　　　　　　　　　　　　
櫻井遊, 梶本和昭, 原島秀吉, Dementia Japan, 30, 1, 30, 40, 2016, [Invited], [Last author, Corresponding author]
Japanese, Introduction scientific journal

BCG-CWS搭載ナノ粒子を用いた膀胱がん免疫療法剤の開発　　　　　　　　　　　　　　　
中村 孝司, 吹上 雅文, 鈴木 嘉晃, 矢野 郁也, 宮崎 淳, 西山 博之, 赤座 英之, 中山 俊憲, 原島 秀吉, 日本薬学会年会要旨集, 135年会, 4, 84, 84, Mar. 2015
(公社)日本薬学会, Japanese

脂質ナノ粒子の粒径制御のためのマイクロ流体デバイス設計
真栄城正寿, 藤島由佳, 佐藤悠介, 石田晃彦, 谷博文, 原島秀吉, 渡慶次学, 化学とマイクロ・ナノシステム学会研究会講演要旨集, 32nd, 2015

多機能性エンベロープ型ナノ構造体の創世とナノ医療への展開　　　　　　　　　　　　　　　
佐藤悠介, 山田勇磨, 梶本和昭, 原島秀吉, 化学とマイクロ・ナノシステム, 14, 1, 15, 23, 2015, [Invited], [Last author, Corresponding author]
Japanese, Introduction scientific journal

Anti-angiogenic nanotherapy via active targeting systems to tumors and adipose tissue vasculature
Yu Sakurai, Kazuaki Kajimoto, Hideyoshi Harashima, BIOMATERIALS SCIENCE, 3, 9, 1253, 1265, 2015, [Peer-reviewed]
English, Book review

Microfluidic approach for production of lipid nanoparticles-based nano medicine
M. Maeki, T. Saito, Y. Node, Y. Sato, T. Yasui, N. Kaji, A. Ishida, H. Tani, Y. Baba, H. Harashima, M. Tokeshi, MicroTAS 2015 - 19th International Conference on Miniaturized Systems for Chemistry and Life Sciences, 838, 840, 2015, [Peer-reviewed]
English, Introduction international proceedings

ヒアルロン酸受容体を用いた肝類洞内皮細胞へ集積する新規Drug delivery system製剤の開発　　　　　　　　　　　　　　　
佐野 直樹, 田村 孝史, 中山 健, 田野井 智倫, 村田 聡一郎, 鳥谷部 尚之, 兵藤 守, 原島 秀吉, 大河内 信弘, 日本DDS学会学術集会プログラム予稿集, 30回, 169, 169, Jul. 2014
日本DDS学会, Japanese

肝臓集積性DDSおよび肝類洞内皮細胞選択性DDSを用いた肝炎治療効果の検討　　　　　　　　　　　　　　　
田野井 智倫, 田村 孝史, 中山 健, 佐野 直樹, 村田 聡一郎, 鳥谷部 尚之, 櫻井 遊, 原島 秀吉, 大河内 信弘, 日本DDS学会学術集会プログラム予稿集, 30回, 178, 178, Jul. 2014
日本DDS学会, Japanese

Multifunctional Envelope-type Nano Device for non-viral gene delivery: Concept and Application for Nanomedicine
Hideyoshi Harashima, JOURNAL OF GENE MEDICINE, 16, 7-8, 204, 205, Jul. 2014
English, Summary international conference

INNOVATIVE TECHNOLOGY FOR IN VIVO MITOCHONDRIAL GENE DELIVERY USING HYDRODYNAMIC LIMB VEIN INJECTION
Yukari Yasuzaki, Yuma Yamada, Tsutomu Kanefuji, Dexi Liu, Hideyoshi Harashima, JOURNAL OF GENE MEDICINE, 16, 7-8, 275, 275, Jul. 2014
English, Summary international conference

【先端的医薬品等におけるレギュラトリーサイエンス研究の現状・課題】 ナノテクノロジーを基盤とした医薬品のレギュラトリーサイエンス研究への取り組み　　　　　　　　　　　　　　　
原島 秀吉, 秋田 英万, 加藤 くみ子, 石井 武彦, 松村 保広, 片岡 一則, Drug Delivery System, 29, 3, 217, 225, Jul. 2014
日本DDS学会, Japanese

Development of Nanotechnology for Mitochondrial Drug Delivery
山田 勇磨, 原島 秀吉, 化學工業, 65, 6, 418, 423, Jun. 2014
小峰工業出版, Japanese

肝臓におけるDrp1の発現抑制は糖代謝を改善する　　　　　　　　　　　　　　　
井林 雄太, 畠山 浩人, 佐藤 悠介, 王 麗華, 原島 秀吉, 高柳 涼一, 野村 政壽, 日本内分泌学会雑誌, 90, 1, 348, 348, Apr. 2014
(一社)日本内分泌学会, Japanese

肝類洞内皮細胞を標的としたDrug Delivery Systemによる新規肝炎治療法の開発　　　　　　　　　　　　　　　
田野井 智倫, 田村 孝史, 村田 聡一郎, 鳥谷部 尚之, 櫻井 遊, 原島 秀吉, 大河内 信弘, 肝臓, 55, Suppl.1, A329, A329, Apr. 2014
(一社)日本肝臓学会, Japanese

BCG-CWS搭載ナノ粒子を基盤とした膀胱癌治療剤の開発
中村 孝司, 吹上 雅文, 中谷 彰洋, 矢野 郁也, 宮崎 淳, 西山 博之, 赤座 英之, 伊藤 俊宏, 細川 裕之, 中山 俊憲, 原島 秀吉, 泌尿器外科, 27, 3, 338, 339, Mar. 2014
医学図書出版(株), Japanese

プラスミドDNA特異的ヒストン修飾による導入遺伝子発現上昇
西原実香, 神田元紀, 神田元紀, 山門振一郎, 原島秀吉, 原島秀吉, 紙谷浩之, 紙谷浩之, 紙谷浩之, 日本DDS学会学術集会プログラム予稿集, 30th, 2014

特異的ヒストン修飾による導入プラスミドDNAの発現上昇
西原実香, 神田元紀, 神田元紀, 山門振一郎, 原島秀吉, 原島秀吉, 紙谷浩之, 紙谷浩之, 紙谷浩之, 紙谷浩之, 日本分子生物学会年会プログラム・要旨集(Web), 37th, 2014

B型肝炎ウイルスの持続感染を再現する効率的な培養細胞評価系の開発に関する研究 自然免疫認識機構の制御によるHBV複製への影響
高岡晃教, 佐藤精一, LI Kai, 亀山武志, 林隆也, 原島秀吉, 秋田英万, 櫻井遊, B型肝炎ウイルスの持続感染を再現する効率的な培養細胞評価系の開発に関する研究 平成25年度 総括・分担研究報告書, 55‐59, 2014
Japanese

ブロック共重合体ミセル医薬品の評価
加藤 くみ子, 中西 健, 小崎 雅人, 松田 嘉弘, 平野 舞, 花田 博幸, 久田 茂, 小野寺 博志, 西山 伸宏, 原島 秀吉, 松村 保広, 片岡 一則, 奥田 晴宏, 川西 徹, 医薬品医療機器レギュラトリーサイエンス, 44, 12, 968, 975, Dec. 2013
(一財)医薬品医療機器レギュラトリーサイエンス財団, Japanese

Global Picture of Nanomedicine Regulation and Promotion
SAKURAGI Makoto, ANZAI Tomohiro, KIMURA Hiromichi, HARASHIMA Hideyoshi, 薬剤学 = The archives of practical pharmacy, 73, 5, 312, 320, 01 Sep. 2013
Japanese

Nano Packagingが拓く次世代核酸医療
松尾保隆, 加地範匡, 畠山浩人, 渡慶次学, 小暮健太朗, 馬場嘉信, 原島秀吉, 表面, 51, 5, 227-240, May 2013, [Peer-reviewed]
Japanese, Report scientific journal

DNAマイクロアレイとin vivo siRNA送達システムの融合による新規2型糖尿病治療薬創出　　　　　　　　　　　　　　　
末光 永理奈, 林 泰弘, 梶本 和昭, 佐藤 悠介, Akhter Afsana, 櫻井 遊, 畠山 浩人, 兵藤 守, 加地 範匡, 馬場 嘉信, 原島 秀吉, 日本薬剤学会年会講演要旨集, 28年会, 237, 237, Apr. 2013
(公社)日本薬剤学会, Japanese

網羅的遺伝子発現情報に基づいた遺伝子キャリアの免疫応答メカニズムの解析　　　　　　　　　　　　　　　
畠山 浩人, 山本 桃子, 林 泰弘, 梶本 和昭, 加地 範匡, 馬場 嘉信, 原島 秀吉, 日本薬学会年会要旨集, 133年会, 4, 83, 83, Mar. 2013
(公社)日本薬学会, Japanese

革新的医薬品の創出に向けたレギュラトリーサイエンス(RS)の推進 ナノテクノロジーを基盤とした革新的医薬品に関する評価方法　　　　　　　　　　　　　　　
原島 秀吉, 松村 保広, 片岡 一則, 日本薬学会年会要旨集, 133年会, 1, 132, 132, Mar. 2013
(公社)日本薬学会, Japanese

樹状細胞に対するKALA修飾MENDの免疫活性化能評価
三浦尚也, 秋田英万, 石井聡一郎, シャリフ シャヒーン, 原島秀吉, 日本薬学会年会要旨集(CD-ROM), 133rd, 2013

樹状細胞に対するKALA修飾MENDの免疫活性化能評価
三浦尚也, 秋田英万, 石井聡一郎, SHARIF Shaheen, 原島秀吉, 医療薬学フォーラム講演要旨集, 21st, 2013

樹状細胞に対するKALA修飾MENDの免疫活性化能評価
三浦尚也, 秋田英万, 石井聡一郎, シャヒーン シャリフ, 中村孝司, 原島秀吉, 日本DDS学会学術集会プログラム予稿集, 29th, 2013

In vivoにおける外来遺伝子発現の持続を達成するDNA配列の同定
神田元紀, 神田元紀, 神田元紀, 小林三和子, 松岡一郎, 原島秀吉, 原島秀吉, 紙谷浩之, 紙谷浩之, 紙谷浩之, 日本分子生物学会年会プログラム・要旨集(Web), 36th, 2013

遺伝子治療用ナノ構造体精製のためのフリーフロー電気泳動マイクロデバイスの開発
重中大輔, 鵜川真美, 加地範匡, 渡慶次学, 秋田英万, 原島秀吉, 馬場嘉信, 日本化学会講演予稿集, 93rd, 2, 2013

Medical Innovation via Pharmaceutical Technologies and Sciences
HARASHIMA Hideyoshi, 薬剤学 = The archives of practical pharmacy, 73, 1, 1, 1, 01 Jan. 2013
Japanese

High throughput purification devices for in vivo applications of gene-delivery multifunctional envelope-type nanodevices
Noritada Kaji, Daisuke Shigenaka, Masami Ukawa, Manabu Tokeshi, Hidetaka Akita, Hideyoshi Harashima, Yoshinobu Baba, 17th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2013, 2, 1170, 1172, 2013, [Peer-reviewed]
English, Summary national conference

膵島移植とミトコンドリア標的型drug delivery system(DDS)
山田勇磨, 野村政壽, 原島秀吉, 人工臓器(日本人工臓器学会), 41, 3, 212, 214, 15 Dec. 2012
JAPANESE SOCIETY FOR ARTIFICIAL ORGANS, Japanese

A Potential non-viral vector to transfect dendritic cell and thereby MHC-Class I antigen presentation might be a potential use in carcinoma
S. Shaheen, H. Akita, I. Souichirou, N. Miura, H. Harashima, CANCER RESEARCH, 72, Dec. 2012
English, Summary international conference

Pretreatment of Hepatocyte Growth Factor Gene Transfer Mediated by Octaarginine Peptide-Modified Nanoparticles Ameliorates LPS/D-Galactosamine-Induced Hepatitis
Yasuhiro Hayashi, Ryoichi Mizuno, Khalil A. Ikramy, Hidetaka Akita, Hideyoshi Harashima, NUCLEIC ACID THERAPEUTICS, 22, 5, 360, 363, Oct. 2012
English

Immunotherapy of bladder cancer with BCG cell wall skeleton encapsulated sub-200 nm nano particles
T. Nakamura, M. Fukiage, M. Higuchi, A. Nakaya, I. Yano, J. Miyazaki, A. Joraku, H. Akaza, T. Ito, H. Hosokawa, T. Nakayama, H. Harashima, IMMUNOLOGY, 137, 763, 763, Sep. 2012
English, Summary international conference

Design of Artificial Gene Carrier Based on the Quantitative Comparison of Intracellular Trafficking with Adenovirus
AKITA Hidetaka, HARASHIMA Hideyoshi, 高分子, 61, 8, 533, 535, 01 Aug. 2012
Japanese

Mitochondrial-targeted DNA delivery using a DF-MITO-Porter, an innovative nano carrier with cytoplasmic and mitochondrial fusogenic envelopes
Yuma Yamada, Eriko Kawamura, Hideyoshi Harashima, JOURNAL OF NANOPARTICLE RESEARCH, 14, 8, 1, 15, Aug. 2012
English

Size-controlled, dual-ligand modified liposomes that target the tumor vasculature show promise for use in drug-resistant cancer therapy
Kazuhiro Takara, Hiroto Hatakeyama, Golam Kibria, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima, JOURNAL OF CONTROLLED RELEASE, 162, 1, 225, 232, Aug. 2012
English

学会報告記 第28回日本DDS学会学術集会を振り返って
原島 秀吉, 秋田 英万, Drug delivery system : DDS : official Journal of the Japan Society of Drug Delivery System, 27, 3, 218, 220, Jul. 2012
日本DDS学会, Japanese

A Multifunctional Envelope-type Nanodevice for Use in Nanomedicine: Concept and Applications
T. Nakamura, H. Akita, Y. Yamada, H. Hatakeyama, H. Harashima, ACCOUNTS OF CHEMICAL RESEARCH, 45, 7, 1113, 1121, Jul. 2012
English, Book review

Intracellular stability of 2'-OMe-4'-thioribonucleoside modified siRNA leads to long-term RNAi effect
Mayumi Takahashi, Chisato Nagai, Hiroto Hatakeyama, Noriaki Minakawa, Hideyoshi Harashima, Akira Matsuda, NUCLEIC ACIDS RESEARCH, 40, 12, 5787, 5793, Jul. 2012
English

ミトコンドリア標的型核酸デリバリーシステムの開発
古川亮, 山田勇磨, 原島秀吉, 日本DDS学会学術集会プログラム予稿集, 28th, 134, 05 Jun. 2012
Japanese

骨格筋ミトコンドリアを標的とした外来遺伝子送達技術の開発
安崎友香理, 山田勇磨, 兼藤努, LIU Dexi, 原島秀吉, 日本DDS学会学術集会プログラム予稿集, 28th, 119, 05 Jun. 2012
Japanese

肺血管内皮細胞標的型siRNAキャリアの標的メカニズムとその抗腫瘍効果　　　　　　　　　　　　　　　
楠本 憲司, 秋田 英万, 松本 有, 野本 貴大, 片岡 一則, 原島 秀吉, 日本DDS学会学術集会プログラム予稿集, 28回, 119, 119, Jun. 2012
日本DDS学会, Japanese

マイクロ流体チップによる多機能性エンベロープ型ナノ構造体の作製　　　　　　　　　　　　　　　
加地 範匡, 北添 雄眞, 重中 大輔, 小暮 健太朗, 秋田 英万, 原島 秀吉, 渡慶次 学, 馬場 嘉信, 日本DDS学会学術集会プログラム予稿集, 28回, 182, 182, Jun. 2012
日本DDS学会, Japanese

Development of a new DNA aptamer against primary cultured tumor endothelial cells by a cell-based SELEX method to discover a new drug delivery system
Mst Naznin Ara, Mamoru Hyodo, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima, CANCER RESEARCH, 72, Apr. 2012
English, Summary international conference

ミトコンドリアを標的とした遺伝子治療に向けて
古川亮, 山田勇磨, 原島秀吉, 日本薬学会年会要旨集, 132nd, 1, 246, 05 Mar. 2012
Japanese

ボンクレキン酸搭載MITO‐Porterの構築およびミトコンドリア送達・抗アポトーシス効果の検証
中村宏平, 山田勇磨, 古川亮, 河村恵理子, 奥田勝博, 新藤充, 原島秀吉, 日本薬学会年会要旨集, 132nd, 4, 209, 05 Mar. 2012
Japanese

Translation of multifunctional envelope-type nano device to nanomedicine
林 泰弘, 梶本 和昭, 原島 秀吉, 細胞, 44, 2, 56, 59, Feb. 2012
ニューサイエンス社, Japanese

クロマチン化プラスミドDNAはin vivoにおいて外来遺伝子発現を上昇させる
紙谷浩之, 紙谷浩之, 紙谷浩之, 紙谷浩之, 宮本紫甫, 後藤仁美, 神田元紀, 神田元紀, 小林三和子, 松岡一郎, 原島秀吉, 原島秀吉, 日本分子生物学会年会プログラム・要旨集(Web), 35th, 2012

クロマチン化プラスミドDNAによる発現上昇
紙谷浩之, 紙谷浩之, 紙谷浩之, 紙谷浩之, 宮本紫甫, 後藤仁美, 神田元紀, 神田元紀, 小林三和子, 松岡一郎, 原島秀吉, 原島秀吉, 日本DDS学会学術集会プログラム予稿集, 28th, 2012

フリーフロー電気泳動法を用いた遺伝子治療用ナノデバイスの精製
重中大輔, 鵜川真美, 加地範匡, 岡本行広, 渡慶次学, 秋田英万, 原島秀吉, 馬場嘉信, 日本化学会講演予稿集, 92nd, 2, 2012

DDS技術による遺伝子・核酸医薬の将来　　　　　　　　　　　　　　　
梶本和昭, 兵藤守, 小暮健太朗, 原島秀吉, 月刊ファインケミカル 特集 先端医療実用化のための最新DDS技術の動向, 41, 4, 31, 36, 2012, [Invited]
Japanese, Introduction scientific journal

多機能性エンベロープ型ナノ構造体の創薬への応用　　　　　　　　　　　　　　　
林泰弘, 梶本和昭, 原島秀吉, 細胞. 特集 疾患治療とドラッグデリバリーシステム, 44, 8, 11, 2012, [Invited]
Japanese, Introduction scientific journal

ドラッグデリバリーシステムの新展開
原島 秀吉, 松村 保広, 木村 廣道, 片岡 一則, HUMAN SCIENCE, 23, 1, 4, 12, Jan. 2012
(公財)ヒューマンサイエンス振興財団, Japanese

P-091 Influences of reduced Werner syndrome protein level on mutagenesis by 8-hydroxyguanine(Poster Presentation)
Kamiya Hiroyuki, Yamazaki Daiki, Harashima Hideyoshi, Taikai Program Yoshisyu of the Environmental Mutagen Society of Japan, 0, 40, 138, 138, 20 Oct. 2011
The Environmental Mutagen Society of Japan, Japanese

A DNA MICROARRAY-BASED ANALYSIS OF THE HOST RESPONSE TO A NONVIRAL GENE CARRIER: EXPLORING CPG-DEPENDENT/INDEPENDENT MECHANISMS
H. Hatakeyama, M. Yamamoto, Y. Hayashi, K. Kajimoto, H. Akita, N. Kaji, Y. Baba, H. Harashima, NUCLEIC ACID THERAPEUTICS, 21, 5, A34, A34, Oct. 2011
English, Summary international conference

IN VIVO SIRNA DELIVERY WITH PH- SENSITIVE LIPOSOME
Yusuke Sato, Hiroto Hatakeyama, Yu Sakurai, Mamoru Hyodo, Hidetaka Akita, Hideyoshi Harashima, NUCLEIC ACID THERAPEUTICS, 21, 5, A41, A42, Oct. 2011
English, Summary international conference