Nov. 2024, 日本ミトコンドリア学会, 第1回 日本ミトコンドリア学会奨励賞　　　　　　　　　　　　　　　
山田勇磨

Jun. 2024, 18th Liposome Research Days 2024, Liposome Research Days Poster Award　　　　　　　　　　　　　　　
Targeting mitochondria based on mitochondrial DDS for cancer therapy
山田勇磨

Nov. 2023, 公益社団法人 コーセーコスメトロジー研究財団, コーセーコスメトロジー奨励賞　　　　　　　　　　　　　　　
山田勇磨

May 2023, 日本薬剤学会, 日本薬剤学会旭化成創剤開発技術賞　　　　　　　　　　　　　　　
山田勇磨

Mar. 2022, The American Pharmacists Association, Ebert Prize Award　　　　　　　　　　　　　　　
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

Jul. 2018, 日本核酸医薬学会奨励賞　　　　　　　　　　　　　　　
山田勇磨

May 2018, 日本酸化ストレス学会学術奨励賞　　　　　　　　　　　　　　　
山田勇磨

Feb. 2017, 北海道科学技術奨励賞（知事表彰）　　　　　　　　　　　　　　　
山田勇磨

2017, Int. J. Pharm.: Elsevier 2017 Outstanding Reviewer　　　　　　　　　　　　　　　

2016, Biomaterials: Elsevier 2016 Outstanding Reviewer　　　　　　　　　　　　　　　

2016, J. Control. Release: Elsevier 2016 Outstanding Reviewer　　　　　　　　　　　　　　　

Jan. 2016, インテリジェント材料・システムシンポジウム 高木賞　　　　　　　　　　　　　　　
山田勇磨

Jul. 2015, 日本DDS学会奨励賞　　　　　　　　　　　　　　　
山田勇磨

May 2014, 日本薬剤学会奨励賞　　　　　　　　　　　　　　　
山田勇磨

Mar. 2014, 日本薬学会奨励賞　　　　　　　　　　　　　　　
山田勇磨

2014, J. Control. Release: Elsevier 2014 Outstanding Reviewer　　　　　　　　　　　　　　　
山田勇磨

Oct. 2011, NINA PHARM ISANH Award　　　　　　　　　　　　　　　
Yuma Yamada

Jun. 2011, 第27回日本ＤＤＳ学会 優秀発表者賞　　　　　　　　　　　　　　　
山田勇磨

Mar. 2006, 日本薬剤学会永井財団大学院学生スカラシップ賞　　　　　　　　　　　　　　　
山田勇磨

Dec. 2005, Asian Mitochondrial Young Investigator Award　　　　　　　　　　　　　　　
山田勇磨

May 2004, PSWC 2004 Student Travel grant　　　　　　　　　　　　　　　
山田勇磨

Cubic Phase-Inducible Zwitterionic Phospholipids Improve the Functional Delivery of mRNA.
Kazuki Iwakawa, Rikako Sato, Mariko Konaka, Yuma Yamada, Hideyoshi Harashima, Yusuke Sato
Advanced science (Weinheim, Baden-Wurttemberg, Germany), 12, 17, e2413016, May 2025, [International Magazine]
English, Scientific journal, Lipid nanoparticles (LNPs) are clinically advanced delivery systems for RNA. The extensively developed structure of ionizable lipids greatly contributes to the functional delivery of mRNA. However, endosomal escape is one of the severe biological barriers that continue to render this process inefficient (e.g., less than 10%). Although LNPs contain phospholipids, their role is poorly understood, and there have been few attempts to perform the chemical engineering required to improve their functionality. Herein, a cubic phase-inducible fusogenic zwitterionic phospholipid derived from 1,2-dioleoyl-3-sn-glycero-phosphoethanolamine (DOPE), DOPE-Cx is described, that is designed to correct this problem. The orientation of a zwitterionic head group of DOPE is engineered by attaching a series of hydrophobic moieties for zwitterionic intermolecular interaction with the head structure of phosphatidylcholine (PC), and this is followed by a lipid-phase transition into non-lamellar phases to facilitate membrane fusion-mediated endosomal escape. A structure-activity relationship study reveals that DOPE-Cx lipids with small hydrophobic chains induce cubic phases instead of a hexagonal phase when mixed with PC, which enhances the functional delivery of mRNA in the liver as opposed to the action of the typically utilized and naturally occurring phospholipids. Engineered functionalized phospholipids will be of great value for the therapeutic applications of mRNAs.

An effective approach to modulate mitochondrial function in murine primary macrophages by a mitochondria-targeted nanocapsule, MITO-Porter.
Mitsue Hibino, Tobia Filosi, Libia Lara Carrion, Emilio Porcu, Ankit Malhotra, Kerstin Lüdtke-Buzug, Saleh M Ibrahim, Yuma Yamada, Misa Hirose
Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, 186, 118019, 118019, May 2025, [International Magazine]
English, Scientific journal, Macrophages play crucial roles in various pathological conditions as well as maintenance of homeostasis. Many chronic inflammatory conditions, such as atherosclerosis, rheumatoid arthritis, and obesity, are known to involve the polarization of macrophages into a proinflammatory state. Therefore, controlling the function of macrophages is a potential strategy to intervene in such pathological conditions. Modulation of immune cell metabolism has recently received attention as a novel therapeutic strategy to counteract such conditions. Recently, a unique nanocapsule (MITO-Porter) that can deliver macromolecules specifically into mitochondria was generated, and its application to improve mitochondrial function was achieved by the direct action of the molecules at the site of the mitochondria in a wide range of cell types but not in immune cells. Therefore, we initiated this study by investigating the feasibility of mitochondria-targeted delivery of coenzyme Q10 (CoQ10), a known antioxidant and cofactor of mitochondrial oxidative phosphorylation, in primary murine bone marrow macrophages (BMDMs) and then evaluated the functional consequences of the treatment with MITO-Porter on mitochondrial function in BMDMs. At steady state, CoQ10-loaded MITO-Porter containing octaarginine (R8) was successfully delivered into the mitochondria, resulting in significant antioxidant effects and increased mitochondrial respiration. Furthermore, the effect of CoQ10 on mitochondrial function in BMDMs was more pronounced when CoQ10 was encapsulated in R8(+) MITO-Porter than when CoQ10 was added alone. This proof-of-concept study highlights the potential of the application of MITO-Porter in macrophages and other immune cells as a novel immunomodulatory therapy for chronic inflammatory conditions.

Splenic B cell-targeting lipid nanoparticles for safe and effective mRNA vaccine delivery.
Yuichi Suzuki, Mai Yakuwa, Mina Sato, Eleni Samaridou, Moritz Beck-Broichsitter, Masatoshi Maeki, Manabu Tokeshi, Yuma Yamada, Hideyoshi Harashima, Yusuke Sato
Journal of controlled release : official journal of the Controlled Release Society, 382, 113687, 113687, 03 Apr. 2025, [International Magazine]
English, Scientific journal, mRNA-loaded lipid nanoparticles (LNPs) have emerged as a potent and versatile platform that underpins the success of mRNA vaccines, but guidelines for designing safe and effective formulations with minimal off-target effects remain unclear. In this study, we focused on a rational design for a novel ionizable lipid library that is based on ionizable tri-oleoyl-tris (iTOT) compounds with a high yield via a simple 2-step synthesis. To further enhance the efficacy and safety of this potent library for vaccine applications, we identified the optimal composition for a vaccine by focusing on the molar ratio of specific lipid excipients in the formulation. This composition brought about a shift in delivery to the spleen, and the LNP formulation, which contained 15 mol% DSPC (15%DSPC-LNPs), was thoroughly taken up by both B cells and other splenic immune cells. This formulation requires neither additional lipid components nor targeting ligand modifications, and it is accompanied by antigen-specific cytotoxic T lymphocyte responses. The rigid, hydrophobic, and charge-neutral surface of 15%DSPC-LNPs minimizes apolipoprotein E-dependent hepatic uptake and maximizes complement receptor-mediated B-cell targeting. Furthermore, as an intramuscularly administered vaccine, 15%DSPC-LNPs induce antigen-specific immune responses and, importantly, results in significantly lower levels of hepatotoxicity compared with that of the mRNA vaccine formulations currently being marketed. In summary, this study demonstrated how the passive targeting of mRNA-LNPs to organs and cells could be regulated by designing novel ionizable lipids combined with adjusting the relative proportions of lipid components. The results of this study also emphasize how selective mRNA delivery to the spleen could avoid the liver, which highlights a promising strategy for the development of safe and effective vaccines.

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.

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

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, 367, 486, 499, Mar. 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.

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

Different Effects of Berberine Delivery to Mitochondria on Cells Derived from the Neural Crest.
Ikuma Hori, Hideyoshi Harashima, Yuma Yamada
Biological & pharmaceutical bulletin, 47, 10, 1726, 1733, 2024, [Domestic magazines]
English, Scientific journal, Energy metabolism is crucial for cell polarity and pathogenesis. Mitochondria, which are essential for maintaining energy homeostasis within cells, can be targeted by drug delivery to regulate energy metabolism. However, there is a lack of research comparing how mitochondria control energy metabolism in different cell types derived from the neural crest. Understanding the effects of berberine (BBR), a compound that acts on mitochondria, on energy metabolism in neural crest-derived cells is important. This study reports how MITO-Porter, a mitochondria-targeted liposome, affects neuroblasts (Neuro2a cells) and normal human epidermal melanocytes (NHEMs) when loaded with BBR. We found that treatment with MITO-Porter containing BBR reduced mitochondrial respiration in Neuro2a cells, while it caused a slight increase in NHEMs. Additionally, the treatment shifted the ATP production pathway in Neuro2a cells to rely more on glycolysis, while in NHEMs, there was a slight decrease in the reliance on glycolysis. We also observed a significant decrease in ATP production in Neuro2a cells, while NHEMs showed a tendency to increase ATP production. Importantly, on the basis of the results of the Premix WST-1 assay, the study found that BBR treatment was not toxic to either cell type. It is important to take note of the varied effects of BBR treatment on different cell types derived from the neural crest. These findings necessitate attention when utilizing NHEMs as a cell model in the development of therapeutic strategies for neurodegenerative diseases, including the use of BBR for metabolic control.

Activation of Mitochondria in Mesenchymal Stem Cells by Mitochondrial Delivery of Coenzyme Q10.
Yuji Maruo, Masahiro Shiraishi, Mitsue Hibino, Jiro Abe, Atsuhito Takeda, Yuma Yamada
Biological & pharmaceutical bulletin, 47, 8, 1415, 1421, 2024, [Domestic magazines]
English, Scientific journal, The efficacy of mesenchymal stem cell (MSC) transplantation has been reported for various diseases. We previously developed a drug delivery system targeting mitochondria (MITO-Porter) by using a microfluidic device to encapsulate Coenzyme Q10 (CoQ10) on a large scale. The current study aimed to confirm if treatment with CoQ10 encapsulated by MITO-Porter enhanced mitochondrial functions in MSCs, with the potential to improve MSC transplantation therapy. We used highly purified human bone marrow-derived MSCs, described as rapidly expanding clones (RECs), and attempted to control and increase the amount of CoQ10 encapsulated in the MITO-Porter using microfluidic device system. We treated these RECs with CoQ10 encapsulated MITO-Porter, and evaluated its cellular uptake, co-localization with mitochondria, changes in mitochondrial respiratory capacity, and cellular toxicity. There was no significant change in mitochondrial respiratory capacity following treatment with the previous CoQ10 encapsulated MITO-Porter; however, mitochondrial respiratory capacity in RECs was significantly increased by treatment with CoQ10-rich MITO-Porter. Utilization of a microfluidic device enabled the amount of CoQ10 encapsulated in MITO-Porter to be controlled, and treatment with CoQ10-rich MITO-Porter successfully activated mitochondrial functions in MSCs. The MITO-Porter system thus provides a promising tool to improve MSC cell transplantation therapy.

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.

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

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, [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.

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.

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.

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.

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.

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.

[Development of an Online Role-play-based Medical Interview Training Method for Fourth-year Pharmacy Students].
Ayako Mori, Izumi Kato, Hitoshi Kashiwagi, Shungo Imai, Katsuya Narumi, Yuki Sato, Ayako Furugen, Yuma Yamada, Masaki Kobayashi
Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan, 142, 6, 661, 674, 2022, [Domestic magazines]
Japanese, Scientific journal, With the coronavirus disease 2019 pandemic, businesses are rapidly expanding their online practices, and the online medical care system has been established and is growing. The field of pharmacy education is also looking for ways to conduct practical online training. Hence, we developed an online role-play-based medical interview training method for fourth-year pharmacy students. The purpose of this study was to describe in detail this method and to clarify the effect of online on medical interviewing practice. The training sessions were conducted using video teleconferencing software. Two settings were used for the role-play scenarios: the pharmacy and hospital. To evaluate the effectiveness of the sessions, a questionnaire was sent to the students, and the results were analyzed using text mining. The most important requirement for successfully conducting the interviews was a stable voice connection, and we reduced audio interruptions and delays by connecting the host personal computer to a wired local area network. We also solved the problem of howling when multiple terminals were installed in the same room by muting all devices in the room. Results of the analysis of the questionnaires suggested that students were more tense online. We also found that students perceived a difference between online and face-to-face interviews in terms of eye contact and the presentation of documents. In this way, we succeeded in conducting smooth online role-playing sessions while taking countermeasures against infection. In the future, it will be necessary to devise nonverbal communication methods and digital methods of presenting the training material.

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

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, [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.

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, [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.

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, 20 Dec. 2020, [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.

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, [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.

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, [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.

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, [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.

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, [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.

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

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.

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.

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], [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, 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], [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.

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, [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.

A mitochondrial delivery system using liposome-based nanocarriers that target myoblast cells.
Takashi Katayama, Shintaro Kinugawa, Shingo Takada, Takaaki Furihata, Arata Fukushima, Takashi Yokota, Toshihisa Anzai, Mitsue Hibino, Hideyoshi Harashima, Yuma Yamada
Mitochondrion, 49, 66, 72, Nov. 2019, [Peer-reviewed], [International Magazine]
English, Scientific journal, Mitochondrial function is reduced in skeletal muscles of many patients with systemic diseases and it is difficult to deliver medicinal substances to mitochondria in such tissue. In this study, we report on attempts to develop liposome-based carriers for mitochondrial delivery using mouse myoblasts (C2C12) by varying the lipid composition of the carriers. We found that a liposome that contains an optimal lipid modified with the KALA peptide (a cellular uptake and mitochondrial targeting device) was the most effective nanocarrier for achieving mitochondrial delivery in C2C12 cells. We also report on successful mitochondrial transgene expression using the carriers encapsulating a mitochondrial DNA vector as we previously reported.

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], [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]

The Use of a Microfluidic Device to Encapsulate a Poorly Water-Soluble Drug CoQ₁₀ in Lipid Nanoparticles and an Attempt to Regulate Intracellular Trafficking to Reach Mitochondria.
Hibino M, Yamada Y, Fujishita N, Sato Y, Maeki M, Tokeshi M, Harashima H
Journal of pharmaceutical sciences, 108, 8, 2668, 2676, Aug. 2019, [Peer-reviewed]
Scientific journal

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.

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]
English, Scientific journal

オートファジー誘導能を有するミトコンドリアDDSの構築およびオートファジー誘導機構の検証　　　　　　　　　　　　　　　
山田 勇磨, 大工原 伸之輔, 西田 慶, 田村 篤志, 由井 伸彦, 原島 秀吉
日本薬学会年会要旨集, 139年会, 4, 59, 59, (公社)日本薬学会, Mar. 2019, [Peer-reviewed]
Japanese

Future of human mitochondrial DNA editing technologies
N. Verechshagina, N. Nikitchina, Y. Yamada, Н. Harashima, M. Tanaka, K. Orishchenko, I. Mazunin
Mitochondrial DNA Part A, 30, 2, 214, 221, Informa UK Limited, 17 Feb. 2019
Scientific journal

[A Nanocarrier System for Mitochondrial Delivery Targeted to a Pancreatic Beta Cell].
Yamada Y
Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan, 139, 1, 41, 45, 2019, [Peer-reviewed]

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

Optimization of siRNA delivery to target sites: issues and future directions.
Khalil IA, Yamada Y, Harashima H
Expert opinion on drug delivery, 1, 13, Sep. 2018, [Peer-reviewed]

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]

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, (株)北隆館, Jul. 2018
Japanese

ポリロタキサン封入MITO-Porterの調製とオートファジーの誘導評価　　　　　　　　　　　　　　　
田村 篤志, 西田 慶, 由井 伸彦, 山田 勇磨, 大工原 伸之輔, 原島 秀吉
生体医歯工学共同研究拠点成果報告書, 平成29年度, 120, 120, 東京医科歯科大学生体材料工学研究所, Apr. 2018, [Peer-reviewed]
Japanese

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]
English, In book

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]
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]
English, Scientific journal

[Analysis by Using Roter Method of Interaction Process Analysis (RIAS) of the Ability of Pharmacy Students to Communicate after Clinical Training for Pharmacy].
Yoh Takekuma, Ayako Mori, Masaki Kobayashi, Yuma Yamada, Yuki Sato, Katsuya Narumi, Ayako Furugen, Mitsuru Sugawara
Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan, 138, 12, 1579, 1586, 2018, [Peer-reviewed], [Domestic magazines]
Japanese, Scientific journal, Communication education is now necessary for pharmaceutical education since the role of pharmacists has expanded from "medicine-based" to "person-based". However, a standard for assessing the effectiveness of a communication education program has not been established. Hence, the aim of this study was to determine the effectiveness of clinical training in pharmacy for enhancing the ability of pharmacy students to communicate. Role playing with simulated patients was performed by pharmacy students before and after clinical practice for pharmacy, and the effects of learning were analyzed by Roter method of interaction process analysis (RIAS). Analysis by RIAS enabled quantification and objective evaluation of communication by pharmacy students. The results showed improvement of interactive communication, decrease of "Question asking" and "Others" including "Transition words", and increase of "Partnership behaviors" and "Counsel behaviors". The pharmacy students became skillful in communication without showing hesitation. The results therefore showed that clinical training contributes to improvement in the ability of pharmacy students to communicate.

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

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

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

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

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

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

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

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

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

Mitochondrial DDS Opens Innovative Pharmaceutics
Yuma Yamada
YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, 136, 1, 55, 62, Jan. 2016, [Peer-reviewed]
Japanese

Mitochondrial dds has a significant impact on the medical and life sciences: The 7th young researcher’s award in the 31th annual meeting of the japan society of drug delivery system
Yuma Yamada
Drug Delivery System, 30, 4, 348, 362, Japan Society of Drug Delivery System, 25 Dec. 2015, [Peer-reviewed]
Japanese, 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

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

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

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

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

Development of a New In Vivo Optical Probe for Biological Diagnosis and Therapy
Ozaki M, Ozawa T, Yamada Y, Nakao K, Minato N, Uemoto S
2015, [Peer-reviewed]

A Post-Liver Transplant Girl With Recurrent Cramps in the Legs.
Abe J, Yamada Y, Suzuki T, Shimamura T, Kobayashi I
Global pediatric health, 2, 2333794X14566074, 2015, [Peer-reviewed], [International Magazine]
English, Scientific journal

p62/SQSTM1 Plays a Protective Role in Oxidative Injury of Steatotic Liver in a Mouse Hepatectomy Model
Sanae Haga, Takeaki Ozawa, Yuma Yamada, Naoki Morita, Izuru Nagashima, Hiroshi Inoue, Yuka Inaba, Natsumi Noda, Riichiro Abe, Kazuo Umezawa, Michitaka Ozaki
ANTIOXIDANTS & REDOX SIGNALING, 21, 18, 2515, 2530, Dec. 2014, [Peer-reviewed]
English, Scientific journal

Development of the MITO-Porter, a Nano Device for Mitochondrial Drug Delivery via Membrane Fusion
Yuma Yamada
YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, 134, 11, 1143, 1155, Nov. 2014, [Peer-reviewed]
Japanese

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

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

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

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

細胞・臓器移植における基礎的研究の最前線 光を応用した移植細胞機能・細胞環境のモニタリングと制御の試み
尾崎 倫孝, 芳賀 早苗, 野田 なつみ, 森田 直樹, 山田 勇磨, 小澤 岳昌
日本外科学会雑誌, 115, 臨増2, 168, 168, (一社)日本外科学会, Mar. 2014
Japanese

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

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

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

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

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]

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

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

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

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

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

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

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, 15, 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

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

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

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

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

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

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

Dual Function MITO-Porter, a Nano Carrier Integrating Both Efficient Cytoplasmic Delivery and Mitochondrial Macromolecule Delivery
Yuma Yamada, Ryo Furukawa, Yukari Yasuzaki, Hideyoshi Harashima
MOLECULAR THERAPY, 19, 8, 1449, 1456, Aug. 2011, [Peer-reviewed]
English, Scientific journal

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

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

Transgene expression efficiency from plasmid DNA delivered as a complex with histone H3
Hiroyuki Kamiya, Hitomi Goto, Genki Kanda, Yuma Yamada, Hideyoshi Harashima
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 392, 1-2, 249, 253, Jun. 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

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

Development of small, homogeneous pDNA particles condensed with mono-cationic detergents and encapsulated in a multifunctional envelope-type nano device
Ryosuke Suzuki, Yuma Yamada, Hideyoshi Harashima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 31, 6, 1237, 1243, Jun. 2008, [Peer-reviewed]
English, Scientific journal

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

MITO-Porter: A liposome-based carrier system for delivery of macromolecules into mitochondria via membrane fusion.
Yamada Y, Akita H, Kamiya H, Kogure K, Yamamoto T, Shinohara Y, Yamashita K, Kobayashi H, Kikuchi H, Harashima H
Biochimica et biophysica acta, 1778, 2, 423, 432, 2, Feb. 2008, [Peer-reviewed]

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.

Efficient cytoplasmic protein delivery by means of a multifunctional envelope-type nano device
Ryosuke Suzuki, Yuma Yamada, Hideyoshi Harashima
Biological and Pharmaceutical Bulletin, 30, 4, 758, 762, 4, Apr. 2007, [Peer-reviewed]
English, Scientific journal

Mitochondrial drug delivery and mitochondrial disease therapy - An approach to liposome-based delivery targeted to mitochondria
Yuma Yamada, Hidetaka Akita, Kentaro Kogure, Hiroyuki Kamiya, Hideyoshi Harashima
MITOCHONDRION, 7, 1-2, 63, 71, Feb. 2007, [Peer-reviewed]
English

Development of multifunctional envelope type artificial viral like gene delivery system
Hideyoshi Harashima, Kentaro Kogure, Yuma Yamada, Hidetaka Akita, Hiroyuki Kamiya
YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, 127, 10, 1655, 1672, 2007, [Peer-reviewed]
Japanese

Significant and prolonged antisense effect of a multifunctional envelope-type nano device encapsulating antisense oligodeoxynucleotide
Y Nakamura, K Kogure, Y Yamada, S Futaki, H Harashima
JOURNAL OF PHARMACY AND PHARMACOLOGY, 58, 4, 431, 437, Apr. 2006, [Peer-reviewed]
English, Scientific journal

Mitochondrial delivery of mastoparan with transferrin liposornes equipped with a pH-sensitive fusogenic peptide for selective cancer therapy
Y Yamada, Y Shinohara, T Kakudo, S Chaki, S Futaki, H Kamiya, H Harashima
INTERNATIONAL JOURNAL OF PHARMACEUTICS, 303, 1-2, 1, 7, Oct. 2005, [Peer-reviewed]
English, Scientific journal

Development of efficient packaging method of oligodeoxynucleotides by a condensed nano particle in lipid envelope structure
Y Yamada, K Kogure, Y Nakamura, K Inoue, H Akita, F Nagatsugi, S Sasaki, T Suhara, H Harashima
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 28, 10, 1939, 1942, Oct. 2005, [Peer-reviewed]
English, Scientific journal

Preparation of a novel PEG-clay hybrid as a DDS material: Dispersion stability and sustained release profiles
T Takahashi, Y Yamada, K Kataoka, Y Nagasaki
JOURNAL OF CONTROLLED RELEASE, 107, 3, 408, 416, Oct. 2005, [Peer-reviewed]
English, Scientific journal

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

ミトコンドリアを標的とするナノカプセル型機能性化粧品の開発
山田勇磨, コスメトロジー研究報告, 32, 2024

Toward the development of nanocapsules that treat mitochondria
山田勇磨, 医学のあゆみ, 291, 6, 2024

Development of delivery technology for targeting mitochondria and approaches to cell therapy applications
山田勇磨, 生化学, 96, 5, 2024

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

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

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, Aug. 2021
日本バイオイメージング学会, 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, Jun. 2021
日本DDS学会, 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, Jun. 2021
日本DDS学会, Japanese

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, Mar. 2021
(公社)日本整形外科学会, 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, Mar. 2021
(公社)日本薬学会, Japanese

オンライン会議システムを活用した医療面接実習の実践　　　　　　　　　　　　　　　
森 綾子, 加藤 いづみ, 柏木 仁, 今井 俊吾, 鳴海 克哉, 佐藤 夕紀, 古堅 彩子, 山田 勇磨, 小林 正紀, 日本薬学会年会要旨集, 141年会, 29V11, pm04, Mar. 2021
(公社)日本薬学会, Japanese

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

Fundamental study on preparation of polymer nanoparticles by microfluidic device
飯岡真吾, 日比野光恵, 山田勇磨, 真栄城正寿, 渡慶次学, 照喜名孝之, 金沢貴憲, 近藤啓, 日本薬学会年会要旨集(Web), 141st, 2021

ミトコンドリア標的型ナノ医薬品の研究開発
山田 勇磨, 薬剤学: 生命とくすり, 80, 6, 280, 285, Nov. 2020
(公社)日本薬剤学会, Japanese

MITO-Porter Technologyに基づいたmitochondrial nano medicine開発への挑戦(The challenge of developing mitochondrial nano medicine based on MITO-Porter Technology)　　　　　　　　　　　　　　　
山田 勇磨, 日本小児循環器学会雑誌, 36, Suppl.2, s2, 103, Nov. 2020
(NPO)日本小児循環器学会, English

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, Nov. 2020
(NPO)日本小児循環器学会, 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
(株)メディカルドゥ, Japanese

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, Oct. 2020
(一社)日本癌学会, English

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, 36回, 163, 163, Aug. 2020
日本DDS学会, Japanese

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, Aug. 2020
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
(株)北隆館, 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, May 2020
(公社)日本薬剤学会, 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
日本バイオマテリアル学会, Japanese

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, Mar. 2020
(公社)日本薬学会, Japanese

【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
(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

ミトコンドリアを標的とする遺伝子治療用ナノカプセルの創製　　　　　　　　　　　　　　　
山田 勇磨, 日本核酸医薬学会誌, 23, 1, 12, 21, Jun. 2019
日本核酸医薬学会, Japanese

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, May 2019
(公社)日本薬剤学会, 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
(株)診断と治療社, Japanese

ミトコンドリア標的型DDSを基盤とした革新的ナノ医薬品の創製
山田 勇磨, ファルマシア, 55, 4, 330, 334, Apr. 2019
(公社)日本薬学会, Japanese

薬学実務実習前後における薬学生のコミュニケーション分析 RIAS(Roter method of interaction process analysis)を用いて　　　　　　　　　　　　　　　
武隈 洋, 森 綾子, 小林 正紀, 山田 勇磨, 佐藤 夕紀, 鳴海 克哉, 古堅 彩子, 菅原 満, 日本薬学会年会要旨集, 139年会, 4, 212, 212, Mar. 2019
(公社)日本薬学会, Japanese

実務実習前後における薬学生のRIASによるコミュニケーション分析　　　　　　　　　　　　　　　
森 綾子, 武隈 洋, 小林正紀, 山田勇磨, 佐藤夕紀, 鳴海克哉, 古堅彩子, 菅原 満, 総合技術研究会2017東京大学, 08 Mar. 2017
Japanese, Summary national conference

Research history of mitochondrial drug delivery system
山田 勇磨, 医学のあゆみ, 260, 1, 42, 48, 07 Jan. 2017
医歯薬出版, Japanese

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]
Japanese, Introduction scientific journal

MENDで拓く遺伝子治療への道 (特集 Drug delivery system(DDS)の最新展望)
佐藤 悠介, 中村 孝司, 山田 勇磨, 医薬ジャーナル, 50, 7, 83, 87, Jul. 2014
医薬ジャーナル社, Japanese

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

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

PD-1-1 光を応用した移植細胞機能・細胞環境のモニタリングと制御の試み(PD-1 パネルディスカッション(1)細胞・臓器移植における基礎的研究の最前線,第114回日本外科学会定期学術集会)
尾崎 倫孝, 芳賀 早苗, 野田 なつみ, 森田 直樹, 山田 勇磨, 小澤 岳昌, 日本外科学会雑誌, 115, 2, 168, 168, 05 Mar. 2014
一般社団法人日本外科学会, Japanese

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

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
Japanese, Book review

医療現場で新しい薬剤師職能を開拓する活動(病院オペ室での活動)と教員としての研究教育活動
小林正紀, 山田勇磨, 佐藤夕紀, 井関健, 日本医療薬学会年会講演要旨集, 22nd, 221, 221, 10 Oct. 2012
日本医療薬学会, 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, Aug. 2012
English

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

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

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

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
English

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
English

手術部サテライトファーマシーにおける麻酔薬の無菌混合調製体制の確立
山崎浩二郎, 須田範行, 新沼朋美, 水口貴史, 小林正紀, 山田勇磨, 山下美妃, 柴山良彦, 山田武宏, 井関健, 井関健, 日本薬学会年会要旨集, 132nd, 4, 218, 218, 05 Mar. 2012
(公社)日本薬学会, Japanese

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

ミトコンドリア標的型DDSが拓くミトコンドリア創薬への道
山田勇磨, 日本薬学会年会要旨集, 132nd, 1, 158, 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

Targeting Mitochondria: Innovation from Mitochondrial Drug Delivery System (DDS) to Mitochondrial Medicine
Yamada Yuma, Harashima Hideyoshi, YAKUGAKU ZASSHI, 132, 10, 1111, 1118, 2012
Mitochondrial dysfunction has been implicated in a variety of human diseases, including cancer and neurodegenerative disorders. Effective medical therapies for such diseases will ultimately require the targeted delivery of therapeutic agents to mitochondria. This will likely be achieved through innovations in the areas of the nanotechnology of intracellular trafficking. Mitochondrial delivery systems for a variety of cargoes have been repored to date. However, only a limited number of approaches are available for delivering macromolecules directly to mitochondria. We previously reported o..., The Pharmaceutical Society of Japan

Drug Delivery System and Mitochondria
YAMADA Yuma, 薬剤学 = The archives of practical pharmacy, 72, 1, 53, 57, 01 Jan. 2012
Japanese

Dual Function MITO-Porter, a Nano Carrier Integrating Both Efficient Cytoplasmic Delivery and Mitochondrial Macromolecule Delivery
Yuma Yamada, Ryo Furukawa, Yukari Yasuzaki, Hideyoshi Harashima, MOLECULAR THERAPY, 19, 8, 1449, 1456, Aug. 2011
English

ミトコンドリア転写因子TFAM‐プラスミドDNA複合体のin vitro転写評価
古川亮, 山田勇磨, 松島雄一, 後藤雄一, 原島秀吉, Drug Deliv Syst, 26, 3, 317, 28 May 2011
Japanese

ミトコンドリア遺伝子を標的とした機能分子送達の試み
山田勇磨, 原島秀吉, Drug Deliv Syst, 26, 3, 285, 28 May 2011
Japanese

ミトコンドリア(Mt)を標的とした遺伝子治療への挑戦―Dual Function MITO‐Porterを用いたMtマトリクスへの核酸送達―
山田勇磨, 河村恵理子, 安崎友香理, 古川亮, 原島秀吉, 日本薬剤学会年会講演要旨集, 26th, 102, 01 May 2011
Japanese

新たなパッケージングコンセプトに基づく多機能性エンベロープ型ナノ構造体の創製
山田勇磨, 鈴木亮佑, 原島秀吉, 日本薬学会年会要旨集, 131st, 4, 159, 05 Mar. 2011
Japanese

ミトコンドリア標的型DDS,MITO‐Porterを用いたオリゴ核酸送達の試み
河村恵理子, 山田勇磨, 原島秀吉, 日本薬学会年会要旨集, 131st, 4, 159, 05 Mar. 2011
Japanese

AN INNOVATIVE APPROACH FOR MITOCHONDRIAL GENE DELIVERY: MITOCHONDRIAL DELIVERY OF OLIGO DNA USING MITO-PORTER, A LIPOSOME-BASED CARRIER FOR MITOCHONDRIAL DELIVERY VIA MEMBRANE FUSION
KAWAMURA Eriko, YAMADA Yuma, HARASHIMA Hideyoshi, 日本ミトコンドリア学会年会要旨集, 11th, 178, 2011
English

TECHNICAL APPLICATION OF HYDRODYNAMIC LIMB VEIN INJECTION FOR IN VIVO MITOCHONDRIAL GENE DELIVERY
YASUZAKI Yukari, YAMADA Yuma, KANEFUJI Tsutomu, LIU Dexi, HARASHIMA Hideyoshi, 日本ミトコンドリア学会年会要旨集, 11th, 170, 2011
English

EFFECT OF DNA PACKAGED WITH TFAM ON TRANSCRIPTION ACTIVATION
FURUKAWA Ryo, YAMADA Yuma, MATSUSHIMA Yuichi, GOTO Yu‐ichi, HARASHIMA Hideyoshi, 日本ミトコンドリア学会年会要旨集, 11th, 179, 2011
English

MITOCHONDRIAL GENOME TARGETING STRATEGY: MITOCHONDRIAL MATRIX DELIVERY OF BIO-MACROMOLECULE USING DF-MITO-PORTER, AN INNOVATIVE NANO CARRIER WITH CYTOPLASMIC AND MITOCHONDRIAL FUSOGENIC ENVELOPES.
YAMADA Yuma, HARASHIMA Hideyoshi, 日本ミトコンドリア学会年会要旨集, 11th, 149, 2011
English

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
English

ボンクレキン酸(抗アポトーシス作動薬)搭載ミトコンドリア標的型DDS(MITO‐Porter)の開発
山田勇磨, 河村恵理子, 安崎友香理, 新藤充, 原島秀吉, Drug Deliv Syst, 25, 3, 307, 28 May 2010
Japanese

DDS技術を基盤とした生細胞ミトコンドリアDNAの可視化
安崎友香理, 山田勇磨, 原島秀吉, 薬剤学, 70, Supplement, 104, 20 Apr. 2010
Japanese

効率的な細胞内導入を目的としたレシチン化SOD(PC‐SOD)搭載R8リポソームの構築およびその機能評価
古川亮, 山田勇磨, 武永美津子, 五十嵐理慧, 原島秀吉, 薬剤学, 70, Supplement, 103, 20 Apr. 2010
Japanese

プラスミド‐ヒストン複合体からの外来遺伝子発現効率
紙谷浩之, 紙谷浩之, 後藤仁美, 後藤仁美, 神田元紀, 神田元紀, 山田勇磨, 山田勇磨, 原島秀吉, 原島秀吉, 薬剤学, 70, Supplement, 148, 148, 20 Apr. 2010
(公社)日本薬剤学会, Japanese

革新的遺伝子発現ベクター『ポリロタキサン/pDNAナノ粒子搭載型MEND』の構築および核内動態・遺伝子発現活性評価
山田勇磨, 野村卓, 原島秀吉, 山下敦, 上遠野亮, 由井信彦, 日本薬学会年会要旨集, 130th, 4, 198, 05 Mar. 2010
Japanese

PC‐SOD搭載型リポソームの構築および細胞内動態・抗酸化効果の評価
古川亮, 山田勇磨, 武永美津子, 五十嵐理慧, 原島秀吉, 日本薬学会年会要旨集, 130th, 4, 198, 05 Mar. 2010
Japanese

生分解性ポリロタキサンを用いたpDNA,siRNAのナノ粒子の調製および核酸ナノ粒子形成機構に関する検討
橋田真裕, 山田勇磨, 原島秀吉, 山下敦, 上遠野亮, 由井信彦, 日本薬学会年会要旨集, 130th, 4, 198, 05 Mar. 2010
Japanese

北海道大学薬学部における実務実習事前実習の取り組みとその評価
武隈洋, 小林正紀, 山田勇磨, 板垣史郎, 吉田和幸, 井関健, 菅原満, 日本薬学会年会要旨集, 130年会, 4, 346, 346, Mar. 2010
(公社)日本薬学会, Japanese

生細胞ミトコンドリアDNAセンシング技術の開発
安崎友香理, 山田勇磨, 原島秀吉, 遺伝子・デリバリー研究会シンポジウム要旨集, 10th, P.38, 2010
Japanese

効率的な遺伝子送達を実現する非ウィルス性ベクターの創製
鈴木亮佑, 山田勇磨, 原島秀吉, 遺伝子・デリバリー研究会シンポジウム要旨集, 10th, P.33, 2010
Japanese

MgO,Al₂O₃/Mg₂Si複合材料の組織におけるSiO₂,Al₂O₃粉末粒径の影響
山田勇磨, 小橋眞, 金武直幸, 日本金属学会講演概要, 145th, 296, 15 Sep. 2009
Japanese

Mono‐cationic detergentによるsiRNAナノ粒子パッケージング法の構築
鈴木亮佑, 山田勇磨, 原島秀吉, Drug Deliv Syst, 24, 3, 326, 09 Jun. 2009
Japanese

細胞内分解性ポリロタキサン搭載多機能性エンベロープ型ナノ構造体の構築
野村卓, 山田勇磨, 原島秀吉, 山下敦, 上遠野亮, 由井伸彦, Drug Deliv Syst, 24, 3, 326, 09 Jun. 2009
Japanese

ミトコンドリア標的型ナノデバイス“多重型MITO‐Porter”の製剤化に関する検討
山田勇磨, 古川亮, 安崎友香理, 原島秀吉, Drug Deliv Syst, 24, 3, 302, 09 Jun. 2009
Japanese

核内pDNA放出制御によるトランスフェクション活性の向上
山田勇磨, 野村卓, 原島秀吉, 山下敦, 上遠野亮, 由井伸彦, 高分子学会予稿集(CD-ROM), 58, 1 Disk1, 1PG147, 12 May 2009
Japanese

Development of Small, Homogenous Non-Viral Gene Vector Using Mono-Cationic Detergents
Ryosuke Suzuki, Yuma Yamada, Hideyoshi Harashima, MOLECULAR THERAPY, 17, S52, S52, May 2009
English, Summary international conference

ミトコンドリア標的型ナノデバイス“多重型MITO‐Porter”の有用性の検証
山田勇磨, 古川亮, 安崎友香理, 原島秀吉, 薬剤学, 69, Supplement, 150, 30 Apr. 2009
Japanese

効率的な核内遺伝子放出を可能とする多機能性エンベロープ型ナノ構造体の創製
山田勇磨, 野村卓, 原島秀吉, 山下敦, 上遠野亮, 由井伸彦, 日本薬学会年会要旨集, 129th, 4, 177, 05 Mar. 2009
Japanese

細胞内分解性ポリロタキサンの構造因子が与える遺伝子送達過程への影響
山下敦, 上遠野亮, 由井伸彦, 山田勇磨, 秋田英万, 原島秀吉, 遺伝子・デリバリー研究会シンポジウム要旨集, 9th, P.24, 2009
Japanese

ミトコンドリア標的型ナノデバイス“多重型MITO‐Porter”の創製
山田勇磨, 古川亮, 安崎友香理, 原島秀吉, 日本ミトコンドリア学会年会要旨集, 9th, 86, 2009
Japanese

多重型MITO‐Porterによるミトコンドリアを標的とした薬物送達戦略
山田勇磨, 秋田英万, 紙谷浩之, 原島秀吉, Drug Deliv Syst, 23, 3, 374, 06 Jun. 2008
Japanese

Development of small, homogeneous pDNA particles condensed with mono-cationic detergents and encapsulated in a multifunctional envelope-type nano device
Ryosuke Suzuki, Yuma Yamada, Hideyoshi Harashima, BIOLOGICAL & PHARMACEUTICAL BULLETIN, 31, 6, 1237, 1243, Jun. 2008
English

効率的な遺伝子送達を目指した1遺伝子パッケージング法の構築
鈴木亮佑, 山田勇磨, 原島秀吉, 薬剤学, 68, Supplement, 194, 30 Apr. 2008
Japanese

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-BIOMEMBRANES, 1778, 2, 423, 432, Feb. 2008
English

Development of Multifunctional Envelope Type Artificial Viral like Gene Delivery System
HARASHIMA Hideyoshi, KOGURE Kentaro, YAMADA Yuma, AKITA Hidetaka, KAMIYA Hiroyuki, Journal of the Pharmaceutical Society of Japan, 127, 10, 1655, 1672, 01 Oct. 2007
This review introduces a new concept "Programmed Packaging" to develop a non-viral gene delivery system. Based on this concept, multifunctional envelope type nano devices (MEND) were developed for in vitro, in situ and in vivo conditions. A quantitative study to identify a rate limiting step in intracellular trafficking was also shown between viral and non-viral vectors, which indicated an important role of controlled intranuclear disposition for development a safe and efficient non-viral gene delivery system. This review will provide a future direction of non-viral gene delivery system., The Pharmaceutical Society of Japan, Japanese

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 第7回 日本DDS学会永井賞受賞によせて
原島秀吉, 小暮健太朗, 秋田英万, 山田勇磨, 紙谷浩之, Drug Deliv Syst, 22, 5, 569, 577, 26 Sep. 2007
Japanese

Development of in vivo gene delivery system by multifunctional envelope-type nano device : Awarded with Nagai Prize in 7th Annual Meeting of the Japan Society of Drug Delivery System
HARASHIMA Hideyoshi, KOGURE Kentaro, AKITA Hidetaka, YAMADA Yuma, KAMIYA Hiroyuki, Drug delivery system, 22, 5, 569, 577, 10 Jun. 2007
日本DDS学会, Japanese

細胞内FRET法を用いたMITO‐Porter(ミトコンドリア膜融性リポソーム)の膜融合能評価
山田勇磨, 秋田英万, 紙谷浩之, 小暮健太朗, 篠原康雄, 原島秀吉, Drug Deliv Syst, 22, 3, 315, 30 May 2007
Japanese

多機能性エンベロープ型ナノ構造体(MEND)を用いたタンパク質の効率的な細胞質送達
鈴木亮佑, 山田勇磨, 原島秀吉, 薬剤学, 67, Supplement, 280, 10 May 2007
Japanese

多機能性エンベロープ型ナノ構造体(MEND)を用いたタンパク質の効率的な細胞質送達
鈴木 亮佑, 山田 勇磨, 原島 秀吉, 薬剤学 = Journal of Pharmaceutical Science and Technology, Japan, 67, 0, 10 May 2007
Japanese

Efficient cytoplasmic protein delivery by means of a multifunctional envelope-type nano device
Ryosuke Suzuki, Yuma Yamada, Hdeyoshi Harashima, BIOLOGICAL & PHARMACEUTICAL BULLETIN, 30, 4, 758, 762, Apr. 2007
English

Efficient cytoplasmic protein delivery by means of a multifunctional envelope-type nano device
Ryosuke Suzuki, Yuma Yamada, Hdeyoshi Harashima, BIOLOGICAL & PHARMACEUTICAL BULLETIN, 30, 4, 758, 762, Apr. 2007
English

ミトコンドリア選択的な高分子薬物送達を可能とする薬物送達システムの開発
山田勇磨, 秋田英万, 紙谷浩之, 小暮健太朗, 篠原康雄, 小林英夫, 菊池寛, 原島秀吉, 日本薬学会年会要旨集, 127th, 1, 284, 05 Mar. 2007
Japanese

タンパク質を効率的に細胞質へ送達するナノ構造体の開発
鈴木亮佑, 山田勇磨, 原島秀吉, 日本薬学会年会要旨集, 127th, 3, 87, 05 Mar. 2007
Japanese

Mitochondrial drug delivery and mitochondrial disease therapy - An approach to liposome-based delivery targeted to mitochondria
Yuma Yamada, Hidetaka Akita, Kentaro Kogure, Hiroyuki Kamiya, Hideyoshi Harashima, Mitochondrion, 7, 1-2, 63, 71, Feb. 2007
English, Book review

Effective drug delivery toward mitochondria by combination of mitochondrial membrane fusion system and mitochondrial targeting signal peptide
YAMADA Yuma, AKITA Hidetaka, KAMIYA Hiroyuki, KOGURE Kentaro, SHINOHARA Yasuo, KOBAYASHI Hideo, KIKUCHI Hiroshi, HARASHIMA Hideyoshi, 日本ミトコンドリア学会年会要旨集, 6th, 31, 13 Dec. 2006
English

Report on The First FIP-APSTJ Joint Workshop on Gene Delivery
YAMADA Yuma, HATAKEYAMA Hiroto, 薬剤学 = Journal of Pharmaceutical Science and Technology, Japan, 66, 6, 451, 453, 01 Nov. 2006
Japanese

膜融合を介してミトコンドリアへの高分子薬物送達を可能とする,“MITO‐Porter”の細胞内での膜融合能評価
山田勇磨, 秋田英万, 紙谷浩之, 小暮健太朗, 山本武範, 篠原康雄, 山下菊治, 小林英夫, 菊池寛, 原島秀吉, Drug Deliv Syst, 21, 3, 298, 20 Jun. 2006
Japanese

オリゴデオキシヌクレオチドの機能向上を目指した多機能性エンベロープ型ナノ構造体への封入
小暮健太朗, 小暮健太朗, 中村宣央, 中村宣央, 山田勇磨, 山田勇磨, 原島秀吉, 原島秀吉, 日本薬学会年会要旨集, 126th, 2, 138, 06 Mar. 2006
Japanese

核膜融合に基づく新規核内送達デバイスの構築
工藤亜沙子, 秋田英万, 増田智也, 山田勇磨, 小暮健太朗, 箕浦ありさ, 原島秀吉, 薬剤学, 66, Supplement, 113, 24 Feb. 2006
Japanese

オリゴDNAの多機能性エンベロープ型ナノ構造体へのパッケージングの確立とその機能評価
中村宜央, 小暮健太朗, 山田勇磨, 原島秀吉, 薬剤学, 66, Supplement, 114, 24 Feb. 2006
Japanese

ミトコンドリアを標的とする膜融合性リポソーム,MITO‐Porterを用いた生細胞ミトコンドリアへの高分子薬物送達―共焦点レーザースキャン顕微鏡を用いた細胞内動態観察―
山田勇磨, 秋田英万, 紙谷浩之, 小暮健太朗, 篠原康雄, 小林英夫, 菊池寛, 原島秀吉, 薬剤学, 66, Supplement, 109, 24 Feb. 2006
Japanese

Development of efficient packaging method of oligodeoxynucleotides by a condensed nano particle in lipid envelope structure
Y Yamada, K Kogure, Y Nakamura, K Inoue, H Akita, F Nagatsugi, S Sasaki, T Suhara, H Harashima, BIOLOGICAL & PHARMACEUTICAL BULLETIN, 28, 10, 1939, 1942, Oct. 2005
English

膜融合を介してミトコンドリアへの送達を可能とするリポソーム,“Mitosome”の開発
山田勇磨, 秋田英万, 小暮健太朗, 紙谷浩之, 篠原康雄, 小林英夫, 菊池寛, 原島秀吉, Drug Deliv Syst, 20, 3, 312, 06 Jul. 2005
Japanese

外来DNAによる転写・翻訳効率の新規定量的評価法
山田勇磨, 紙谷浩之, 原島秀吉, 日本薬学会年会要旨集, 125th, 2, 147, 05 Mar. 2005
Japanese

MITO-Porter, a new-type carrier for mitochondrial macromolecules delivery system via a membrane fusion mechanism
YAMADA Yuma, AKITA Hidetaka, KAMIYA Hiroyuki, KOGURE Kentaro, SHINOHARA Yasuo, KOBAYASHI Hideo, KIKUCHI Hiroshi, HARASHIMA Hideyoshi, 日本ミトコンドリア学会年会要旨集, 5th, 69, 2005
English

直接導入法とDNAトランスフェクション法による蛋白質送達の定量的評価
山田勇磨, 紙谷浩之, 秋田英万, 原島秀吉, Drug Deliv Syst, 19, 3, 290, 10 May 2004
Japanese

オルガネラDDSを基盤とする次世代医薬品の開発　　　　　　　　　　　　　　　
山田勇磨
和歌山医科大学 病態解析学研究室 特別講演, Invited oral presentation
2025, [Invited]

Development of mitochondria-targeted nano capsule to transport genome editing devices.　　　　　　　　　　　　　　　
Yuma Yamada
The 27th Plant Organelle Workshop / Grant-in-Aid Transformative Research Areas (A) Cytoplasmic Genome Regulation JOINT SYMPOSIUM, 2025, Nominated symposium

ミトコンドリア標的型DDSを基盤とした創剤研究　　　　　　　　　　　　　　　
山田勇磨
令和６年度 AMED村山班・小坂分担班 集中TR会議, 2025, Invited oral presentation
[Invited]

ミトコンドリア標的型 DDS 技術の社会実装を目指して　　　　　　　　　　　　　　　
山田勇磨
第29回 創剤フォーラム 若手研究会, 2024, Invited oral presentation
[Invited]

オルガネラ標的型DDSが拓く新時代Nano medicine　　　　　　　　　　　　　　　
山田勇磨
第40回日本DDS学会学術集会（若手ワークショップ）, 2024, Nominated symposium
[Invited]

脂質ナノ粒子調製のイロハ　　　　　　　　　　　　　　　
山田勇磨
第40回日本DDS学会学術集会（教育セッション）, 2024, Public discourse
[Invited]

オルガネラ制御を基盤とする細胞操作の実践 学術シンポジウム2: 細胞材料を素材とする創剤研究　　　　　　　　　　　　　　　
山田勇磨
日本薬剤学会第39年会(学術シンポジウム2), 2024, Public symposium

光を届けるDDSの創製を目指して～オルガネラ標的型ＤＤＳを基盤とする癌光治療への挑戦～　　　　　　　　　　　　　　　
山田勇磨
日本薬学会第144年会, 2024, Public symposium

Targeting Mitochondria Based on Mitochondrial Drug Delivery Systems (DDS)　　　　　　　　　　　　　　　
Yuma Yamada
14th edition of the World Congress on Targeting Mitochondria 2023, Oct. 2023
[Invited]

細胞を操るオルガネラ制御学の創出を目指して　　　　　　　　　　　　　　　
山田勇磨
第39回日本DDS学会学術集会（若手ワークショップ）, Jul. 2023
[Invited]

ミトコンドリアを標的とした癌光治療を実践するナノカプセルの創製　　　　　　　　　　　　　　　
山田勇磨
日本薬剤学会第38年会(旭化成創剤開発技術賞受賞講演), May 2023
[Invited]

ミトコンドリア標的型 DDS を基盤としたナノ医療の創出を目指して　　　　　　　　　　　　　　　
山田勇磨, 原島秀吉
第43回 生体膜と薬物の相互作用シンポジウム, 06 Oct. 2022
[Invited]

Validation of therapeutic strategies using mitochondrial drug delivery systems　　　　　　　　　　　　　　　
Yamada Y, Harashima H
Joint Symposium of the Faculty of Pharmaceutical Sciences & WPI-ICReDD in Hokkaido University, 15 Sep. 2022
[Invited]

治療用rRNA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
日本核酸医薬学会第7回年会, Jul. 2022, Japanese, Invited oral presentation
[Invited]

ミトコンドリアDDSを基盤とした遺伝子細胞治療への挑戦　　　　　　　　　　　　　　　
山田勇磨
第28回日本遺伝子細胞治療学会学術集会, Jul. 2022, Japanese, Invited oral presentation
[Invited]

ミトコンドリアDDSの夢： ミトコンドリア標的型ナノ医薬品の創出　　　　　　　　　　　　　　　
山田勇磨
日本薬剤学会第37年会, May 2022, Japanese, Invited oral presentation
[Invited]

ミトコンドリアを標的とする核酸ナノ医薬品の創製を目指して　　　　　　　　　　　　　　　
山田勇磨
ゲノム創薬・創発フォーラム第9回シンポジウム, Feb. 2022, Japanese, Invited oral presentation
[Invited]

ミトコンドリアを標的とする遺伝子治療戦略の検証　　　　　　　　　　　　　　　
山田勇磨
第20回日本ミトコンドリア学会年会, Dec. 2021, Japanese, Invited oral presentation
[Invited]

未来型DDSが拓くミトコンドリアを標的とする新しいナノ医療　　　　　　　　　　　　　　　
山田勇磨
京都大学呼吸器内科セミナー, Nov. 2021, Japanese, Public discourse
[Invited]

Targeting mitochondria: innovation of mitochondrial drug delivery system (DDS) to mitochondrial medicine.　　　　　　　　　　　　　　　
山田勇磨
Mitochondrial Medicine -Therapeutic Development., Nov. 2021, English, Invited oral presentation
[Invited]

オルガネラ標的型DDSを基盤とした核酸医薬開発研究の最前線　　　　　　　　　　　　　　　
山田勇磨
第15回Wakoウェブ受託セミナー, Nov. 2021, Japanese, Public discourse
[Invited]

The way to develop an innovative nanomedicine targeting mitochondria.　　　　　　　　　　　　　　　
山田勇磨
Host Madsen Medal winner 2021 ceremony., Sep. 2021, English, Invited oral presentation
[Invited]

オルガネラ標的型DDSを基盤とした創剤研究の最前線　　　　　　　　　　　　　　　
山田勇磨
第72回北日本小児科学会, Sep. 2021, Japanese, Invited oral presentation
[Invited]

ミトコンドリア機能を操作するDrug Delivery System　　　　　　　　　　　　　　　
山田勇磨
第30回日本バイオイメージング学会, Sep. 2021, Japanese, Invited oral presentation
[Invited]

ミトコンドリア標的型 Drug Delivery System を基盤とした遺伝子治療戦略の検証　　　　　　　　　　　　　　　
山田勇磨
日本ゲノム編集学会第6回大会, Jun. 2021, Japanese, Invited oral presentation
[Invited]

希少疾患の未来について考える〜患者とともに挑むイノベーション　　　　　　　　　　　　　　　
山田勇磨
iPark Frontier Talk, Feb. 2021, Japanese, Invited oral presentation
[Invited]

The challenge of developing mitochondrial nano medicine based on MITO-Porter Technology　　　　　　　　　　　　　　　
山田勇磨
第56回日本小児循環器学会 (JSPCC-AHA 2020), Nov. 2020, English, Invited oral presentation
[Invited]

ミトコンドリア標的型ナノ医薬品の創製を目指して　　　　　　　　　　　　　　　
山田勇磨
第35回日本小児神経学会北海道地方会, Oct. 2020, Japanese, Invited oral presentation
[Invited]

MITO-Porter, a cutting-edge mitochondrial DDS　　　　　　　　　　　　　　　
山田勇磨
Pharmaceutical Society of Korea 2020, Oct. 2020, English, Invited oral presentation
[Invited]

新たな○△□治療を、ミトコンドリアに薬を運ぶ技術開発で!　　　　　　　　　　　　　　　
山田勇磨
TAKEDA Academic WEB Salon2020, Aug. 2020, Japanese, Invited oral presentation
[Invited]

Mito-porter, Investigation of Drug Delivery System Targeting Mitochondria.　　　　　　　　　　　　　　　
山田勇磨
The 84th annual scinetific meeting of the japanse circulation society (JCS2020), Jul. 2020, English, Invited oral presentation
[Invited]

A Mitochondrial Dds Towards An Innovative Therapy.　　　　　　　　　　　　　　　
山田勇磨
2020 Annual Meeting & Exposition of the Controlled Release Society, Jun. 2020, English, Invited oral presentation
[Invited]

光よ届け！ミトコンドリアへ　　　　　　　　　　　　　　　
山田 勇磨
第9回 生体物理化学セミナー, 24 Jan. 2020, Invited oral presentation
[Invited]

ミトコンドリアDDSが描くナノ医療の未来予想図～リポ化プロスタグランジン製剤の話題を含めて～　　　　　　　　　　　　　　　
山田勇磨
第12回 北海道小児オータムセミナー, 23 Nov. 2019, Invited oral presentation
[Invited]

ミトコンドリア標的型ナノＤＤＳを基盤とした遺伝子・細胞治療の実装　　　　　　　　　　　　　　　
山田勇磨
第12回 CHEMBIOハイブリッドレクチャー, 19 Oct. 2019, Invited oral presentation
[Invited]

MITO-Porter, liposomal mitochondrial delivery system: Toward Mitochondrial Nanomedicine. MITO-Porter therapy to open innovative therapies targeting mitochondria.　　　　　　　　　　　　　　　
Yuma Yamada
16th ASMRM & 19th J-mit., 05 Oct. 2019, Invited oral presentation
[Invited]

MITO-Porter, liposomal mitochondrial delivery system: Toward Mitochondrial Nanomedicine.　　　　　　　　　　　　　　　
Yuma Yamada
Liposome Research Days 2019., 18 Sep. 2019, Invited oral presentation
[Invited]

ﾐﾄｺﾝﾄﾞﾘｱDDSを用いた酸化ｽﾄﾚｽ応答制御および疾患治療への展開　　　　　　　　　　　　　　　
山田勇磨
第72回日本酸化ストレス学会, 28 Jun. 2019, Invited oral presentation
[Invited]

ミトコンドリア標的型ナノカプセルが創る未来医療　　　　　　　　　　　　　　　
山田勇磨
IVF JAPANセミナー, 04 Feb. 2019, Invited oral presentation
[Invited]

The MITO-Porter integrates a mitochondrial drug delivery system with a variety of other current scientific concepts with the goal of developing new innovative technologies and medicines.　　　　　　　　　　　　　　　
Yuma Yamada
The 19th RIES-HOKUDAI International Symposium., 11 Dec. 2018, Invited oral presentation
[Invited]

ミトコンドリア標的型DDSを基盤としたナノ医療の創出を目指して　　　　　　　　　　　　　　　
山田勇磨
第18回 日本ミトコンドリア学会年会, 08 Dec. 2018, Invited oral presentation
[Invited]

Mitochondrial DDS for Gene & Cell Therapy.　　　　　　　　　　　　　　　
Yuma Yamada
Cadiovascular and Metabolic Week 2018., 07 Dec. 2018, Invited oral presentation
[Invited]

Mitochondrial DDS opens emerging therapies and novel strategy of new drug development for mitochondrial disorders.　　　　　　　　　　　　　　　
Yuma Yamada
KSAP annual convention 2018., 12 Oct. 2018, Invited oral presentation
[Invited]

ミトコンドリア機能を制御するナノカプセルの構築および疾患治療に向けた試み　　　　　　　　　　　　　　　
山田勇磨
第91回 日本生化学大会, 25 Sep. 2018, Invited oral presentation
[Invited]

ミトコンドリアDDSの使い方　　　　　　　　　　　　　　　
山田勇磨
第9回 フリーラジカルスクール, 04 Aug. 2018, Invited oral presentation
[Invited]

ミトコンドリアを標的とする遺伝子治療用ナノカプセルの創製　　　　　　　　　　　　　　　
山田勇磨
日本核酸医薬学会第4回年会, 09 Jul. 2018, Invited oral presentation
[Invited]

遺伝子・細胞治療に用いるミトコンドリアDDS　　　　　　　　　　　　　　　
山田勇磨
22 Jun. 2018, Invited oral presentation
[Invited]

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
第60回日本小児神経学会学術集会, 31 May 2018, Invited oral presentation
[Invited]

Mitochondrial Nano DDS Toward Innovative Medicine and Therapy.　　　　　　　　　　　　　　　
Yuma Yamada
HIGO Program Cutting edge Seminar., 18 Apr. 2018, Invited oral presentation
[Invited]

膵島ミトコンドリアを標的とした分子送達技術の開発および糖尿病治療への展開　　　　　　　　　　　　　　　
山田勇磨
日本薬学会 第138年会, 28 Mar. 2018, Invited oral presentation
[Invited]

ミトコンドリア病の遺伝子改変治療　　　　　　　　　　　　　　　
山田勇磨
H29 年度 村山班ミトコンドリア病診断・診療マニュアル作成委員拡大合宿, 10 Feb. 2018, Invited oral presentation
[Invited]

ミトコンドリア機能を制御するナノカプセルの開発　　　　　　　　　　　　　　　
山田勇磨
第6回ミトコンドリア機能研究会, 03 Feb. 2018, Invited oral presentation
[Invited]

Nano-drug delivery system for mitochondrial gene therapy.　　　　　　　　　　　　　　　
Yuma Yamada
The 18th International Union of Materials Research Societies, International Conference in Asia (IUMRS-ICA) 2017, 08 Nov. 2017, Invited oral presentation
[Invited]

ミトコンドリアＤＤＳの開発と医療分野への展開　　　　　　　　　　　　　　　
山田勇磨
第11回 次世代を担う若手医療薬科学シンポジウム, 22 Oct. 2017, Invited oral presentation
[Invited]

ミトコンドリア標的型ナノカプセルMITO-Porterを基盤とした遺伝子治療への挑戦　　　　　　　　　　　　　　　
山田勇磨
第59回日本先天代謝異常学会, 12 Oct. 2017, Invited oral presentation
[Invited]

Validation of mitochondrial gene therapy using a MITO-Porter　　　　　　　　　　　　　　　
Yuma Yamada
8th SFRR-Asia & 14th ASMRM., 09 Sep. 2017, Invited oral presentation
[Invited]

ミトコンドリアDDSを基盤とした遺伝子・細胞治療への展開　　　　　　　　　　　　　　　
山田勇磨
第33回日本DDS学会, 06 Jul. 2017, Invited oral presentation
[Invited]

ミトコンドリアを診る・操る・治すDrug Delivery System　　　　　　　　　　　　　　　
山田勇磨
第2回先端ケミカルバイオロジー研究会, 06 Jun. 2017, Invited oral presentation
[Invited]

ミトコンドリア標的型ナノカプセル MITO-Porter　　　　　　　　　　　　　　　
山田勇磨
H28 年度 村山班ミトコンドリア病診断・診療マニュアル作成委員拡大合宿, 11 Feb. 2017, Invited oral presentation
[Invited]

ミトコンドリアDDSの創製と心疾患治療への展開　　　　　　　　　　　　　　　
山田勇磨
第3回iHFフォーラム, 07 Aug. 2016, Invited oral presentation
[Invited]

ミトコンドリア標的型ナノマシン“MITO-Porter”の開発　　　　　　　　　　　　　　　
山田勇磨
第24回日本Cell Death学会, 29 Aug. 2015, Invited oral presentation
[Invited]

MITO-Porter, an innovative technology for mitochondrial drug delivery.　　　　　　　　　　　　　　　
Yuma Yamada
The 1st HU-TMU-KU Joint Symposium for Pharm. Sci., 29 Aug. 2015, Invited oral presentation
[Invited]

ミトコンドリアDDSが拓く医療・ライフサイエンス革命　　　　　　　　　　　　　　　
山田勇磨
第31回日本DDS学会, 11 Jul. 2015, Invited oral presentation
[Invited]

ミトコンドリアDDSの創製とナノ医療への展開　　　　　　　　　　　　　　　
山田勇磨
北海道大学病院・循環器内科主催・循環病態内科学リサーチセミナー, 26 May 2015, Invited oral presentation
[Invited]

ミトコンドリアを科学するナノデバイス MITO-Porter の創製　　　　　　　　　　　　　　　
山田勇磨
日本薬学会 第135年会, 27 Mar. 2015, Invited oral presentation
[Invited]

ミトコンドリア DDS が拓く新しい薬剤学　　　　　　　　　　　　　　　
山田勇磨
日本薬学会 第135年会, 26 Mar. 2015, Invited oral presentation
[Invited]

薬剤学I　　　　　　　　　　　　　　　
北海道大学
2024

-　　　　　　　　　　　　　　　
Hokkaido University

臨床薬学技術実習　　　　　　　　　　　　　　　
北海道大学

実践的薬物送達学コース　　　　　　　　　　　　　　　
北海道大学

医療薬学特論　　　　　　　　　　　　　　　
北海道大学

一般教育演習(ﾌﾚｯｼｭﾏﾝｾﾐﾅｰ)　　　　　　　　　　　　　　　
北海道大学

先端生物科学実験法 II　　　　　　　　　　　　　　　
北海道大学

OSCE対応演習　　　　　　　　　　　　　　　
北海道大学

実務実習事前実習　　　　　　　　　　　　　　　
北海道大学

薬剤学II　　　　　　　　　　　　　　　
北海道大学

オルガネラへの核酸および生理活性分子導入
科学研究費助成事業
01 Apr. 2024 - 31 Mar. 2029
沼田 圭司, 山田 勇磨, Law SimonSauYin
日本学術振興会, 学術変革領域研究(A), 京都大学, 24H02272

Genomic regulation of intracellular symbiotic organelles: from technological innovation to understanding and application of life phenomena
Grants-in-Aid for Scientific Research
01 Apr. 2024 - 31 Mar. 2029
有村 慎一, 石原 直忠, 竹中 瑞樹, 鳥山 欽哉, 松村 浩由, 風間 智彦, 庄司 佳祐, 沼田 圭司, 神吉 智丈, 山田 勇磨, 木内 隆史, 高梨 秀樹, 細川 正人, 佐藤 美由紀, 西村 芳樹, 小笠原 絵美, 矢守 航
Japan Society for the Promotion of Science, Grant-in-Aid for Transformative Research Areas (A), The University of Tokyo, 24H02270

Activated mRNA delivery system based on core-shell structured LNP with smart polymers
Grants-in-Aid for Scientific Research
12 Apr. 2023 - 31 Mar. 2028
原島 秀吉, 佐藤 悠介, 中村 孝司, 山田 勇磨, 西山 伸宏, 内田 智士
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (S), Hokkaido University, 23H05451

小児の心筋症におけるミトコンドリア障害と心筋エネルギー代謝に関する研究
科学研究費助成事業
01 Apr. 2023 - 31 Mar. 2026
武田 充人, 佐々木 大輔, 山田 勇磨
日本学術振興会, 基盤研究(C), 北海道大学, 23K07496

ミトコンドリア遺伝子治療を実現するゲノム編集技術の創出
科学研究費助成事業
01 Apr. 2023 - 31 Mar. 2026
山田 勇磨, 真栄城 正寿, 武田 充人, 佐々木 大輔, 日比野 光恵
日本学術振興会, 基盤研究(A), 北海道大学, 23H00541

脳疾患治療を実現するミトコンドリア標的型ナノカプセルの開発　　　　　　　　　　　　　　　
科学研究費助成事業 挑戦的研究(萌芽)
30 Jun. 2022 - 31 Mar. 2025
山田 勇磨
日本学術振興会, 挑戦的研究(萌芽), 北海道大学, 22K19928

Functional background of mitochondrial genome evolution unraveled from the atp8 gene of Polypedates leucomystax
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
01 Apr. 2022 - 31 Mar. 2025
安岡 有理, 山田 勇磨
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Institute of Physical and Chemical Research, 22K06348

Study on mechanisms of immune and tissue tolerance after allogeneic hematopoietic stem cell transplantation
Grants-in-Aid for Scientific Research
01 Apr. 2021 - 31 Mar. 2025
豊嶋 崇徳, 冨塚 一磨, 山田 勇磨
前年までに示した同種造血幹細胞移植マウスモデルにおけるsingle cell RNA seq(scRNAseq)解析によってカルシニューリン阻害薬(CNI)投与によって誘導されるtransitory exhausted T cell (t-Tex)の詳細を解析した。CNI投与によってドナーT細胞のC1、C10クラスターが増加した。C1クラスターは疲弊関連遺伝子（Tox, Pdcd1など）が低発現である一方エフェクター関連遺伝子(Gzmb, Cx3Cr1, Ly6cなど)が高発現であるCD8＋t-Texであり、C10クラスターはCD4＋t-Texと考えられた。scTCRseq解析ではこれらはoligoclonalであり、CNI投与によるアロ応答性T細胞のクローン性増殖によるものであると考えられた。これらCD8＋t-Tex、CD4＋t-Texに共通する細胞表面マーカーとしてLy6cを同定した。これらはCNI中止後も長期間残存し、stemnessの性格が認められた。またCNI投与後に抗PD-L1抗体を投与したところT細胞機能、殺細胞性が増強し、GVHDの増悪、GVL効果の増強がみられた。これらの結果から、GVHD予防のCNIによってドナーT細胞のアロ応答性が維持され、慢性GVHD、GVL効果を誘導することが示された。またCAR-T細胞の機能低下にフェロトーシスが関与していることを明らかにし、フェロトーシス阻害剤Fer-1をCAR-T製造培養系に添加することでTCF-1+ stem-like CAR-T細胞が増加し、殺細胞活性の増強を認めた。マウスへの投与実験ではより長期の生存が確認できた。臓器特異的GVHD治療開発のための組織特異的デリバリー創薬については、抗HER2ヒト化抗体4D5-scFv、抗マウス/ヒトGPA33ヒト抗体G9-scFv、抗マウス/ヒトEpCAMヒト抗体Ep203-scFvの作成が完了した。
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 23K21439

Development of Phototherapeutic Drugs and Elucidation of Single Molecule Dynamics Using Highly Bright Stable Quantum Dots and Innovative 3D cell system
Grants-in-Aid for Scientific Research
01 Apr. 2021 - 31 Mar. 2025
高野 勇太, 宮武 由甲子, 平田 恵理, 山田 勇磨, 繁富 香織
本年度は、当初計画通り「(II):マイクロ組織内の分子動態解明を利用した高効果光がん治療薬の開発」および「(III):マイクロ組織を活用した光治療薬開発プロセスの一般化」を中心に研究遂行した。また、本研究の遂行から発明された新規ナノ粒子作成方法について「(I):3D培養系で高い浸透性・発光性・安定性をもつ機能化量子ドットの合成と培養組織内動態の解明」に関連して新規展開し、その理解の深化と基盤構築を並行して行った。
(II)に関して、各種光増感剤の改良・新規開発を行った。我々がこれまでも用いているπ拡張型ポルフィリン分子(rTPA)を利用した系の開発に成功した【成果論文2報】。ここではrTPAについて包括的な開発を行い、その液中挙動及び光励起状態での光増感剤としての性能評価をおこない、用途に応じて至適なrTPA誘導体を選択する基盤を確立した。この知見をもとに、ナノ薬剤やナノカーボンとの複合化による光治療薬向け化合物開発が加速した。
(III)に関して、歯科医療応用に向けた近赤外光治療向けの化合物開発を行った。カーボンナノホーンを担体としてrTPAと複合化することで、効果的な近赤外光増感作用を発現して殺菌効果を示す材料開発を達成した【成果論文1報】。
新規展開の(I)について、量子ドット凝集体を利用することでより長時間露光や環境変化に対しても安定に発光をつづける発光ラベルが得られることを見出した（特願2023- 79183、論文準備中）。この知見は、長時間観察が必要になることの多い3次元培養細胞系において、有用な発光ラベル調製法となりため、これを利用した薬物輸送キャリアのプロトタイプ開発も行った。これらは、3次元細胞系を基軸にした展開を行っていた本研究ゆえに実現した物質開発といえる。
以上のように、研究は申請書記載の当初計画案にそっておおむね順調に進行している。
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 23K21067

Development of Phototherapeutic Drugs and Elucidation of Single Molecule Dynamics Using Highly Bright Stable Quantum Dots and Innovative 3D cell system
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
01 Apr. 2021 - 31 Mar. 2025
高野 勇太, 宮武 由甲子, 平田 恵理, 山田 勇磨, 繁富 香織
本年度は、当初計画通り「(I):3D培養系で高い浸透性・発光性・安定性をもつ機能化量子ドットの合成と培養組織内動態の解明（研究１～2年目計画）」のために細胞標的指向性分子-量子ドット複合体の開発に着手した。量子ドットに対してポリエチレングリコール鎖を介して水酸基（中性）およびカルボン酸（アニオン性）を接合した量子ドットを合成した。また、量子ドットに対して癌細胞マーカーとして良く知られているCD44抗体およびEpCAM抗体を接合した複合体も作成し、血液モデルのPBMC中から癌細胞を選択的に癌細胞認識できることを確認した。当該量子ドット-抗体複合体は現在、膵がん細胞における取り込み挙動を、2次元培養および3次元培養にて検討中である。また、量子ドット自体の光安定性についても検討を行い、光照射強度に応じて表面エッチングが起きる一方、光強度の制御や量子ドット表面組成によって細胞毒性を抑え得ることを見出した。
また「(II):マイクロ組織内の分子動態解明を利用した高効果光がん治療薬の開発（研究２～３年目計画）」に関して、その前段階として光増感剤の開発検討を行った。我々がこれまでも研究に用いているπ拡張型ポルフィリン分子(rTPA)について、量子ドット表面への担持やナノカーボン表面への担持を行い、その光増感性能（＝一重項酸素発生能）の検証を行った。その結果、量子ドット表面への直接不可は、分子凝集などが原因となり十分な一重項酸素発生能を発揮しない一方、ナノカーボンの1種であるカーボンナノホーンに担持したrTPAは十分な一重項酸素発生能と殺がん効果を発揮し得ることを見出した。
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 21H01753

造血幹細胞移植後の免疫寛容と組織寛容の機序の解明とその制御法の開発研究
科学研究費助成事業 基盤研究(B)
01 Apr. 2021 - 31 Mar. 2025
豊嶋 崇徳, 冨塚 一磨, 山田 勇磨
マウス同種造血幹細胞移植後にcyclosporin A (CSP)を投与することによって, ドナーT細胞疲弊の進行が停止し, 疲弊T細胞 (Tex)の前駆細胞 (precursor Tex: pTex)が増加することを示した。pTexが保たれることによって, 移植後の免疫チェックポイント阻害剤の抗腫瘍効果が増強された。一方, 移植後のT細胞を回収して, 別のレシピエントに輸注したところ, 1回目の移植後にCSPを利用していると, ２回目の移植後に慢性移植片対宿主病(慢性GVHD)が発症することが示された。一方, 移植後大量シクロフォスファミド（Posttransplant cyclophosphamide: PTCY）法を行ってからCSPを移植後day5から投与しても, pTexは誘導されなかった。CSPによるGVHD予防は長期的には慢性GVHDの発症に繋がる可能性が示された。
移植後にFLT3阻害剤 (gilteritinib)投与によって, 移植後にFLT3変異陽性急性白血病細胞からIL-15の産生が促進され, ドナーT細胞のTexの分化が抑制され, 白血病に対する細胞傷害活性が増強されることを示した。移植後day5からday14まで短期間のgilteritinib投与で, GVHDの増悪無しでGVL効果が増強されて, マウスの生存が延長されることを示し, Bone marrow transplantation誌に発表した。
ミトコンドリアナノメディシンを利用した悪性腫瘍治療法の開発のため, ミトコンドリア外膜の活性を保った単離ミトコンドリア (Q)を利用した。新規キメラ抗原受容体T細胞（CAR-T細胞）作成時にQを加えることによって, CAR-T細胞内にQを取り込ませ, CAR-T細胞の作成効率を改善することが可能となった。Qを利用することによって臨床的に問題となる, CAR-T細胞の作成失敗を防ぐことが期待される。
日本学術振興会, 基盤研究(B), 北海道大学, 21H02944

Development of Trojan horse-type cancer therapeutics by elucidating and utilizing the predation mechanism of pancreatic cancer cells
Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Research (Exploratory)
09 Jul. 2021 - 31 Mar. 2023
高野 勇太, 宮武 由甲子, 山田 勇磨, 繁富 香織
本年度は当初計画通り、「[段階-I] 捕食誘引物質の粗分離、観察、構造決定による同定」にとりかかり、膵がん細胞の死細胞を調製した上で捕食誘引物質の分離を開始した。まず、死細胞について大きく「アポトーシス様死細胞」と「ネクローシス様死細胞」に分けて調製し、それぞれ膵がん細胞より再構築したミクロ組織3次元と共培養しながらリアルタイム顕微鏡観察を行い、取り込まれる挙動の観察を行った。この際、コントロール条件として1μm（大体アポトーシス死細胞と同サイズ）のCOOH修飾蛍光ビーズを用いた。結果、COOH修飾蛍光ビーズでは細胞への取り込み、特に細胞集団が触手を伸ばすなどの能動的な取り込みは起きないことを確認した。そして「アポトーシス様死細胞」と「ネクローシス様死細胞」は両者とも取り込まれることを確認したが、ネクローシス様死細胞では取り込みがやや少ない傾向にあった。これはネクローシスでは死細胞構造がランダムであるとともに、細胞内容物が維持できないことが一因であると考えられた。よって、今後の検討は主にアポトーシス様死細胞で行うこととした。現在は、アポトーシス様死細胞の大量調製と、粗分離を開始したところである。
また、「[段階-II]捕食誘引物質を複合化した高薬効性の光がん治療薬の合成開発」の準備段階として、量子ドットに近赤外光殺がん分子(rTPA)を接合した複合体や、ドラッグデリバリキャリアとrTPAの複合化も行い、その一重項酸素発生能および光殺がん効果を確認した。本研究の遂行によってドラッグデリバリキャリアとrTPAの相性により、内容分子の機能性（本研究の場合は光殺がん性能）が変化することが分かった。今後、最適化を進め最大効率で殺がん効果を示す複合体プロトタイプを作成し、本研究の目的とするトロイの木馬型殺がん化合物を完成させる。
Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Research (Exploratory), Hokkaido University, 21K19036

Production of novel drug derivery system with light-induced disruption of liposomes (LiDL)
Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Research (Exploratory)
30 Jul. 2020 - 31 Mar. 2023
須藤 雄気, 山田 勇磨
【目的】物理化学（代表者）と薬剤学（分担者・協力者）の融合による光誘起崩壊リポソーム：Light-induced Disruption of Liposomes（LiDL）の開発と、それに基づく新奇薬物送達手法の確立。【背景】狙った時間と場所に薬物を届け・働かせることは、薬学における大きな『夢』である。【計画】光受容タンパク質・ロドプシンとpH感受性ポリマーおよび任意の化合物（薬物）を内封させたリポソームを開発することで、時空間制御性に優れた『光』により、狙った時間・場所で薬物を放出させる新奇手法を確立する。【意義】薬学における『夢』の一つを叶える手法となり、大きな波及効果をもたらす。


具体的には、光受容タンパク質「(1) ロドプシン（H+ポンプ・チャネル）」と「(2) pH 感受性ポリマー」を含む「(3) リポソーム」を作成する。その際、「(4) 化合物 (薬物)」を内封させる。このリポソームに「(5) 光」を照射すると、ロドプシンが活性化され、リポソーム内外のpH が大きく（> 5 ユニット）変化する。これにより、pH 感受性ポリマーの物理的形状が変化し、リポソームが崩壊し、化合物が「(6) 放出」される。LiDL と命名するこの手法は、時空間分解能に優れた「光」により薬物を放出させるという、新奇かつ独創性・汎用性の高い薬物送達（DDS）手法になる。本年度は、ロドプシンを組み込んだリポソームの作成と、光によるpH変化を定量的に測定し、ロドプシン組み込みリポソームが狙い通りに機能することを明らかにした。今後は、このリポソームにpH感受性分子を組み込むとともに、光により崩壊するかを内部に導入する蛍光分子の蛍光変化により確認する。
Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Research (Exploratory), Okayama University, 20K21482

Development of a nano device for mitochondrial gene editing
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
01 Apr. 2020 - 31 Mar. 2023
山田 勇磨
ミトコンドリアのゲノム変異と種々の疾患との関連が報告されており、本オルガネラを標的とした遺伝子治療が期待されている。本申請研究では、ミトコンドリア標的型ナノカプセル (MITO-Porter)を用いて標的ミトコンドリアへゲノム編集装置を運搬し、ミトコンドリア機能を遺伝子レベルで治療する事を目的とする。2021年度は、1. ゲノム編集の検証および2. ミトコンドリア機能の評価について、下記に記載した計画で研究を進めた。
1. ゲノム編集の検証： 疾患細胞が保有するtRNA領域のmtDNA変異をゲノム編集可能な装置をパッケージングしたMITO-Porterを調製し、疾患細胞ミトコンドリア内部にゲノム編集装置を導入した。単離したミトコンドリアを用いて、定量的PCR法を利用してミトコンドリア内部のmtDNA変異の切断効率を測定し、ゲノム編集装置搭載MITO-Porterが変異mtDNAを効率的に切断することを確認した。さらに、疾患細胞での評価を行い、キャリアの最適化を図った。
2. ミトコンドリア機能の評価： 実験に用いる疾患細胞は、ミトコンドリア呼吸活性複合体の活性低下が観察されている。そのため、治療効果の指標としてミトコンドリア呼吸活性 （細胞外フラックスアナライザー）を評価する。2022年は、疾患細胞のミトコンドリア呼吸活性を測定する手法を確立した。現在、ゲノム編集装置搭載MITO-Porter投与時の疾患細胞ミトコンドリア呼吸活性を測定し、投与プロトコルの最適化を図っている。
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 20H04523

Innovative gene/nucleic acid delivery system based on optimized intracellular trafficking steps
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)
01 Apr. 2019 - 31 Mar. 2023
原島 秀吉, 佐藤 悠介, 中村 孝司, 山田 勇磨
１ siRNA送達（佐藤）：siRNAは細胞質中でリン酸化酵素Clp1により5’末端のリン酸化修飾を受け、RISCを形成し、標的mRNAのサイレンシングを引き起こす。我々の予備検討結果から、siRNAの多くが上述の一般的な経路をたどっていない可能性が強く示唆された。そこでブラックボックスとなっているsiRNAの細胞内運命を①5’リン酸化・RISC形成速度、②細胞質内・核内局在、の観点から解明する。
２ pDNA送達（原島）：転写(TC)/翻訳(TL)過程に大きな差が認められたナノDDSにおいて、TCとTLのどちらの寄与が大きいかを、mRNAを定量することにより識別する。 ①TLの寄与が大きい場合、ナノDDSのエンドソーム脱出時にエンドソーム内物質が漏出したり、あるいはpDNA導入に伴う各種細胞内センサーの刺激によって、導入細胞におけるTL活性が低下する、という仮説を検証する。
３ がん免疫ナノ療法（中村）：患者個人の腫瘍関連微小環境の免疫状態を把握することは治療戦略の決定に不可欠である。そこで、免疫状態が異なる担がんマウスモデルから腫瘍組織、所属リンパ節、脾臓を採取し、遺伝子発現解析を行うことで腫瘍関連微小環境の免疫状態情報を得る。文献情報からヒトで確認されている腫瘍関連微小環境の免疫状態をカテゴリー化し、それらを反映する担がんマウスモデルを5種類以上選定する。 ４ Mt送達（山田）：核酸・遺伝子をコアとするナノ粒子を形成し、細胞膜とMt膜 を突破するために、脂質膜コーティングを施す。脂質膜表面には細胞導入素子およびミトコンドリア移行性アプタマー(負電荷)を修飾したDual ligand MITO-Porterを基盤骨格とする。また、細胞内での流動性の向上を目指し、ポリエチレングリコール（PEG）などの親水性ポリマーの修飾や、微小な粒子径も検討する。
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (A), Hokkaido University, 19H01170

Development of artificial mitochondria with cell-symbiosis ability
Grants-in-Aid for Scientific Research
30 Jul. 2020 - 31 Mar. 2022
Yamada Yuma
Mitochondria transplantation therapy, in which mitochondria are transplanted into diseased cells, could be a very useful, but there is a problem in that optimal symbiosis (cellular internalization and mitochondrial communication) for therapeutic efficacy has not been achieved. The aim of this research was to develop artificial mitochondria with enhanced symbiotic ability for use in therapeutic applications. This research proposal includes (i) the formulation of artificial mitochondria and the optimization of their intracellular dynamics, (ii) validation of their therapeutic efficiency using a model of mitochondrial disease cells. A number of results regarding the development of artificial mitochondria have been achieved and evaluated in preliminary experiments.
Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Research (Exploratory), Hokkaido University, 20K21872

Establishment of the new diagnostic method in mitochondrial cardiomyopathy
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
01 Apr. 2019 - 31 Mar. 2022
Takeda Atsuhito
Mitochondrial cardiomyopathy (MCM), one of the secondary cardiomyopathies, is diagnosed definitively by pathology, tissue biochemistry, and genetic testing, but there are no clear criteria for myocardial pathology. Therefore, we measured mitochondrial volume density in electron microscopy images and performed immunohistochemistry using respiratory chain enzyme antibodies in MCMs diagnosed definitively by clinical symptoms, genetic testing, and tissue biochemistry, and compared them with other cardiomyopathies other than MCM. Mitochondrial volume density was significantly higher in the MCM group, and immunohistochemistry showed decreased staining for Complex I or IV in the MCM group. This study allowed the pathological diagnosis of MCM using electron microscopy and immunohistochemistry, which is very useful for the pathological criteria of MCM.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 19K08529

Drug discovery of nano-molechule mitochondria targeted agent and impact on post cardiac arrest brain injury
Grants-in-Aid for Scientific Research
01 Apr. 2017 - 31 Mar. 2021
Kohei Ikeda
Mitochondrial dysfunction following ischemia/reperfusion injury including cardiac arrest and CPR is characterized by an impairment of mitochondrial function which leads to pro-apoptotic signaling and cell death. No pharmacological agent has yet been found to improve clinical outcomes after cardiac arest. We developed a new mitochondria targeted nano-particle involving active form vitamin B1, VB1-MITO-Porter. This VB1-MITO-Porter statistically increased ATP turn over of SHSY5Y cell's mitochondria in-vitro. However, VB1-MITO-Porter did not improve 10-day survival and histological brain injury on murine cardiac arrest model.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Jikei University School of Medicine, 17K11063

異常ミトコンドリアを認識する遺伝子治療用ナノカプセルの構築　　　　　　　　　　　　　　　
科学研究費助成事業（基盤研究(B)）
Apr. 2017 - Mar. 2020
山田勇磨
文部科学省, Principal investigator, Competitive research funding

Investigation into unraveling the mechanism of drug delivery system to the inner ear
Grants-in-Aid for Scientific Research
01 Apr. 2016 - 31 Mar. 2019
Kita Tomoko
The final aim of this study was to achieve the effective drug delivery systems to the inner ear. First, we tried to establish the culture system of blood-labyrinth barrier but unfortunately failed. And second, to clarify the contribution of each drug transporter expressed in this inner ear barrier, we have established the preparation protocols of cDNA library from samples obtained by laser capture micro dissection. Third, using several model drugs for each drug transporter and their inhibitors, it was shown that several tissue type specific transporters are localized in the inner ear barrier, but there was no significant change in the distribution of each model drug.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Kyoto University, 16K11180

ミトコンドリアの品質管理を科学するナノデバイスの開発　　　　　　　　　　　　　　　
科学研究費補助金(挑戦的萌芽研究)
Apr. 2017 - Mar. 2019
山田勇磨
文部科学省, Principal investigator, Competitive research funding

Establishment of diagnostic procedure and the new diagnostic criteria of mitochondrial cardiomyopathy based on an Imaging technology
Grants-in-Aid for Scientific Research
01 Apr. 2015 - 31 Mar. 2018
Takeda Atsuhito, ABE JIRO
Mitochondrial disease, which is a designated intractable disease, has not yet been established as a treatment method, and it is urgent to establish a diagnostic method leading to early detection. We have aimed to establish mitochondrial cardiomyopathy diagnostic criteria including mitochondrial cardiomyopathy model and its evaluation system. In the basic experiments, the results were confirmed in the preparation and evaluation system of doxorubicin mitochondrial cardiomyopathy-induced model animals and the rescue experiment using mitochondrial DDS. Clinically, as diagnostic criteria for mitochondrial cardiomyopathy, we identified three points of (1) myocardial pathology tissue: mitochondrial abnormality in electron microscopic image, (2) tissue biochemistry: myocardial tissue respiratory chain enzyme activity reduction, and (3) gene mutation as necessary elements.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 15K09679

Development of optical control method of cell organelle function by photoindeuced charge separation molecule
Grants-in-Aid for Scientific Research
01 Apr. 2015 - 31 Mar. 2018
Takano Yuta, IMAHORI Hiroshi, MURAKAMI Tatsuya, YAMADA Yuma
In this study, novel functional molecules utilizing the photoinduced charge separation were developed for a fundamental technology to develope photocontrol methods of cellular activities. We investigated intracellular optical functionality mainly for amphipathic ferrocene-porphyrin-fullerene linked molecules and aryl-acridinium molecules.
As research outcomes, we succeeded in photoinduced depolarization of the cell membrane by high charge separation efficiency due to development of molecules with enhanced cell membrane fluidity. In addition, we succeeded in mitochondria selective molecular transport in living cells by utilization of drug transport carriers or optimization of molecular structure. The molecules demonstrated successful method for triggering photooxidation/reduction reactions in cells by irradiation with light.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), 15K05563

Elucidation of organelle network mechanism controlled by mitochondrial dynamics and its application.
Grants-in-Aid for Scientific Research
01 Apr. 2014 - 31 Mar. 2017
Nomura Masatoshi, ISHIHARA Naotada, YAMADA Yuma
Mitochondria are highly dynamic organelles that frequently fuse and divide in response to cellular energy demands. In an excessive energy state such as a high fat diet, the organelle network between Mt and ER is impaired, followed by the induction of ER stress. As a result, FGF21 is secreted from the liver by activation of PERK-eIF2a-ATF4 pathway, and energy expenditure is accelerated in the peripheral organs such as skeletal muscle and adipose tissue. In other words, mitochondrial dynamics-ER stress-FGF21 axis function as a biological defense system for energy excess. Therefore, mitochondrial dynamics is a therapeutic target for the obese diabetes.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Kyushu University, 26461383

New approach for the mechanism of ototoxic drugs against the mtDNA1555AG mutation - human iPSC and nanocarieer-
Grants-in-Aid for Scientific Research
01 Apr. 2014 - 31 Mar. 2017
Kitajiri Shinichiro, KITA Tomoko, IKI Takehiro, MIZUKOSHI Akifumi
To clarify the mechanism of ototoxicity with aminoglycoside antibiotice (AG), we have established the experiment systems of ototoxicity for MtDNA1555AG iPSC. In the recent technology, it is impossible to make transgenic mouse for this mitochondrial SNP mutation.
Compared with hair cells, most cells don't have any transport systems for taking AG into the cells. We therefore use nanocarrier encapsulated AG. When we use this nanocarrier for Hela cell line, only mitochondria specific carrier showed cell death dose-dependently. We established patient drived iPSC cell lines and characterized them. We further optimized the composition of nanocarrier for iPSC, and Ag encapsulated nanocarrier are now preparing.
Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Exploratory Research, Kyoto University, 26670741

Development of a nanoparticle to release nucleic acids in a mitochondrial environment
Grants-in-Aid for Scientific Research
2015 - 2016
YAMADA Yuma, HYODO Mamoru
It was recently reported that mutations in mitochondrial DNA (mtDNA) form the basis of mitochondria (Mt) related diseases. Therefore, the delivery of nucleic acids targeting mutant mtDNA would be expected to be an innovative therapeutic strategy for such diseases. To date, we developed a MITO-Porter, a nano carrier for mitochondrial delivery. The aim of this study is to develop a nanoparticle that can release nucleic acids in response to the Mt environment following its delivery to the Mt. In this study, we evaluated nanoparticles with various Mt environment responsive peptides, and we succeeded in constructing a nanoparticle that was capable of releasing nucleic acids in response to conditions that mimic the Mt environment. In addition, we confirmed successful regulation of the mitochondrial gene expression by transfection of nanoparticles using the MITO-Porter. Our findings can contribute to development nanomedicine for mitochondrial gene therapy.
Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Exploratory Research, Hokkaido University, Principal investigator, Competitive research funding, 15K12532

Development of nanomachine for mitochondrial gene therapy targeted to cells derived from mitochondrial disease patients
Grants-in-Aid for Scientific Research
2014 - 2016
YAMADA Yuma, AKITA Hidetaka, HYODO Mamoru
In order to establish a therapeutic strategy targeting genes in mitochondria (Mt), we examined Mt gene delivery and mitochondrial exogenous gene expression in cells derived from Mt disease patients. To date, we have been able to develop a MITO-Porter, a nano carrier for Mt delivery. In this study, we report on the successful Mt gene delivery in disease cells that have a heteroplasmic mutation in mtDNA, using MITO-Porter system. In addition, we designed a pHSP-mtLuc (CGG) analog, an artificial Mt gene expression plasmid DNA vector that contains a promotor for Mt transcription and an artificial Mt genome with a reporter gene that records adjustments to the Mt codon system. Collectively, we succeeded in achieving Mt exogenous gene expression by the Mt delivery of a pHSP-mtLuc (CGG) analog using the developed MITO-Porter system. Our Mt exogenous gene expression carriers promises to be a useful approach to Mt gene therapy and research regarding Mt molecular biology.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, Principal investigator, Competitive research funding, 26282131

MENDで拓く遺伝子治療への道：遺伝子の運び屋からナノマシンへ
科学研究費助成事業
31 May 2010 - 31 Mar. 2014
原島 秀吉, 山田 勇磨, 中村 孝司, 兵藤 守, 馬場 嘉信, 篠原 康雄, 小暮 健太朗, 紙谷 浩之, 渡慶次 学, 松尾 保孝, 秋田 英万
パッチワーク法による新しいMEND構築方法を確立し一枚膜でコア粒子をパッケージする新しい方法を開発し、平均粒子径が25nm程度でゼータ電位が-40mV程度の構造体を得ることに成功し、P-MENDと命名した。D-MEND法と組み合わせることでPD- MEND(3枚膜構造)を構築したところ、市販の試薬より高い活性を示した。マイクロ流体デバイスを開発し、最小粒径30nmのMENDを作製することに成功した。一方でマイクロ流路作製についての検討し、ナノインプリント手法によりガラスマイクロ流路の量産が可能であることをしめした。また、シースフローと電場を利用したマイクロフリーフロー電気泳動デバイスの構造・分離条件の最適化を行い、MEND精製に適用することでMENDの定性的かつ定量的評価法を確立した。KALAペプチドを導入した脂質膜封入型ナノ粒子にがん抗原等をコードした遺伝子を封入することで、抗腫瘍活性を得ることに成功した。また、本粒子は細胞に対し免疫活性化効果を有することを示し、その機構に細胞質内DNAセンサーが関与することを示唆する結果を得た。酵素耐性RNAアプタマーの単離に世界で始めて成功した。Mt移行性ペプチドを搭載したMTS-MITO-Porterを構築し、生細胞内での選択的Mt送達システムを開発し、MTS-MITO-Porterは従来型MITO-Porterと比較してミトコンドリア移行能が飛躍的に上昇した。GAL4とVP16の融合蛋白質を用いる自己活性化システムを構築し、reporterプラスミドDNAの単独投与に比べて数十倍にルシフェラーゼ活性が上昇した。アンチセンスRNA (ASO)をMITO-Porterを用いて細胞Mtに送達し、ミトコンドリアmRNAを40%抑制し、さらに標的mRNAがコードする内因性Mtタンパク質の発現量が低下した。
日本学術振興会, 基盤研究(S), 北海道大学, 22229001

Development of nano device for regulating mitochondrial gene expression
Grants-in-Aid for Scientific Research
2013 - 2014
YAMADA Yuma
Mitochondria have attracted interest as an organelle that can be used as a target in medical therapy, in the maintenance of beauty & health and the development of the life sciences. The objectives of the research supported by this grant is to develop a nano device that can be used for regulating mitochondrial gene expression including mitochondrial gene silencing and mitochondrial transgene expression. In this study, we report on the successful mitochondrial delivery of nucleic acids in human cells using a MITO-Porter, which is a liposome-based carrier that is capable of introducing macromolecular cargos into mitochondria via membrane fusion. Moreover, we achieved mitochondrial RNA knockdown and mitochondrial transgene expression using such a MITO-Porter system. Our system for regulating mitochondrial gene expression system promises to have a significant impact on the medical and life sciences.
Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Exploratory Research, Hokkaido University, Principal investigator, Competitive research funding, 25560219

Development of in vivo mitochondrial gene delivery system and validation of mitochondrial exogenous gene expression in skeletal muscle
Grants-in-Aid for Scientific Research
2011 - 2012
YAMADA Yuma
Mitochondrial genetic disorders are a major cause of mitochondrial diseases including mitochondrial inherited diseases, neurodegenerative diseases and diabetes. It is therefore likely that mitochondrial gene therapy will be useful for the treatment of such diseases. To achieve such an innovative therapy, successful mitochondrial exogenous gene expression, two independent processes, i.e., "development of in vivo mitochondrial gene delivery system" and "construction of DNA vector to achieve mitochondrial gene expression" are required. In this study, we report on successful mitochondrial gene delivery in skeletal muscle using hydrodynamic limb vein (HLV) injection. Moreover, we could construct the DNA vector which can express on mitochondria through hydrodynamic gene delivery. Our mitochondrial exogenous gene expression system promises to be a useful technique for mitochondrial gene therapy and research regarding mitochondrial molecular biology.
Japan Society for the Promotion of Science, Grant-in-Aid for Young Scientists (A), Hokkaido University, Principal investigator, Competitive research funding, 23680053

Development of Multi-layered nano-device for mitochondrial drug delivery and an innovative approach for mitochondrial disease' therapy
Grants-in-Aid for Scientific Research(若手研究(B))
2009 - 2010
Yuma YAMADA
Mitochondrial dysfunction is associated with a variety of human diseases. Effective medical therapies for mitochondrial diseases will ultimately require an optimal drug delivery system, which will likely be achieved through innovations in the nanotechnology of intracellular trafficking. To achieve efficient mitochondrial drug delivery, two independent processes, i.e., "cytoplasmic delivery through the cell membrane" and "mitochondrial delivery through the mitochondrial membrane" are required. Here, we report the development of a Dual Function MITO-Porter (DF-MITO-Porter), a nano carrier int...
Ministry of Education, Culture, Sports, Science and Technology, 若手研究(B), 北海道大学, Principal investigator, Competitive research funding, 21700483

Multifunctional Envelope-type Nano Device as non-viral gene delivery system for cancer therapy
Grants-in-Aid for Scientific Research(特定領域研究)
2008 - 2009
Hideyoshi HARASHIMA, 秋田 英万, 紙谷 浩之, 山田 勇磨, 畠山 浩人, 馬場 嘉信
In this study, we developed a multifunctional envelope-type nano device (MEND) as a non-viral siRNA delivery system for cancer, which can induce the knockdown of tumor specific genes and anti-tumor effect with no toxicity. First, we measured antigen presentation of OVA encapsulated R8-MEND in dendritic cell (DC). As a result, R8-MEND showed specific MHC class I presentation, although showed low MHC class II presentation. Moreover, mice subcutaneously immunized by R8-MEND were showed significant antitumor effect compared with control mice. Next, we prepared GALA/DMEND encapsulating siRNA cor...
Ministry of Education, Culture, Sports, Science and Technology, 特定領域研究, 北海道大学, Coinvestigator not use grants, Competitive research funding, 20015003

抗原提示バランスとアジュバントの制御に基づいたオーダーメイド癌ワクチン
科学研究費補助金(萌芽研究, 挑戦的萌芽研究)
2008 - 2009
原島 秀吉, 小暮 健太朗, 秋田 英万, 山田 勇磨
本年度は、アジュバントのトポロジー制御の観点からアジュバント搭載型ナノ構造体の構築とその機能評価を行った。まず認識受容体の局在が異なる2種類のアジュバントを搭載したMENDの構築を行い、細胞性免疫誘導効率への影響を調べた。polyI:Cはエンドソーム内のTLR3及び細胞質中のMda5により認識され、CpG-ODNはエンドソーム内のTLR9によって認識される。それ故、細胞への取り込み後に認識されるように、MENDの内部に抗原と共に内封した。polyI:CもしくはCpG-ODNを内封したMENDをマウスに皮下免疫し、CTL活性を測定した結果、polyI:C内封MENDを免疫したマウス群では、CpG-ODN搭載MENDを免疫した群と比較して著しく高いCTL活性の誘導が認められた。次にpolyI:C搭載MENDとpolyI:C/R8複合体のCTL活性誘導能を比較した。その結果、polyI:C搭載MENDを免疫したマウス群の方が高いCTL活性及び抗腫瘍活性を示した。MEND内にpolyI:Cを内封することで、効率良くTLR3及びMda5に認識され、強力な細胞性免疫が誘導されたと考えられる。このことからアジュバントの機能を十分に発揮させるためには、そのトポロジーを考慮することが重要であることが示唆された。またpolyI:CをMEND内に内封することでアジュバント投与時の炎症等の副作用が軽...
文部科学省, 萌芽研究, 挑戦的萌芽研究, 北海道大学, Coinvestigator not use grants, Competitive research funding, 20659020

Development of in vivo gene delivery system for siRNA and genome wide screening of type-two diabetes related genes in mice and rats
Grants-in-Aid for Scientific Research(基盤研究(A))
2006 - 2009
Hideyoshi HARASHIMA, 中谷 明弘, Hiroyuki KAMIYA, 小暮 健太朗, 秋田 英万, Yoshinobu BABA, Yuma YAMADA
We performed genome-wide screening of type-2 diabetes related genes which can be detectable in blood cells before disease state or after disease occurred. In parallel, we succeeded in developing in vivo gene delivery system for liver..In addition, we constructed an original assay system for nano carriers to evaluate transcytosis of endothelial cells and finally succeeded in finding a selective ligand for transcytosis of nano carriers.
Ministry of Education, Culture, Sports, Science and Technology, 基盤研究(A), 北海道大学, Coinvestigator not use grants, Competitive research funding, 18200032

Development of Multi-layered nano-device for mitochondrial drugdelivery and mitochondrial protein delivery for cancer therapy
Grants-in-Aid for Scientific Research(若手研究(スタートアップ))
2007 - 2008
Yuma YAMADA
ミトコンドリア(Mt)は様々な疾患と密接に関わっており、Mtを標的とした薬物治療が注目を集めている。これらの治療を実現するためには、Mtへ治療薬物を送達する必要があるが、有用なシステムは報告されていない。本研究では、Mtを標的とした新規送達システムを開発する事に成功し、癌細胞Mtへの薬物送達および薬理効果を確認した。本システムを用いる事で、今までは不可能であった難治性疾患の治療が期待できる。
Ministry of Education, Culture, Sports, Science and Technology, 若手研究(スタートアップ), 北海道大学, Principal investigator, Competitive research funding, 19890001

がん選択的多機能性エンベロープ型ナノ構造体の開発とがん遺伝子治療への応用
科学研究費補助金(特定領域研究)
2006 - 2007
原島 秀吉, 紙谷 浩之, 小暮 健太朗, 秋田 英万, 山田 勇磨
我々は、多機能性エンベロープ型ナノ構造体(MEND)の開発に成功し、細胞膜透過機能を有するオクタアル,ギニン(R8)修飾したR8-MENDは、培養細胞系においてアデノウイルスと同等の遺伝子発現を示すことを明らかにした(Khalil,et al.Gene Ther.2007)。さらに、in vivoでがん組織選択的な遺伝子デリバリーシステムを開発するために、がん組織内で特異的に発現するペプチダーゼ(MMP)により切断されてPEGを脱離し、がん細胞内へ侵入するという新しい戦略(PEG-peptide-DOPE(PPD)-MEND)を設計し,本コンセプトが機能するかをin vitro系で検証した。その結果、PPD-MENDはMMP依存的にPEGが解離し、高い遺伝子導入を誘起できることが明らかとなった。さらに、本PPD-MENDをマウスに静脈内投与すると、血中滞留性を示すと同時に、腫瘍組織へ有為な送達が可能であり、in vivoにおける遺伝子発現にも成功した(Hatakeyama,et al.Gene Ther.2007)。さらに、PPD-MENDにsiRNAを搭載し、in vivoで腫瘍組織へ標的化可能、かつ、がん細胞で発現している遺伝子(ルシフェラーゼ遺伝子)を選択的にノックダウンすることにも成功した。本、PPD-MENDに抗腫瘍効果を誘起するsiRNAを搭載することにより、がん...
文部科学省, 特定領域研究, 北海道大学, Coinvestigator not use grants, Competitive research funding, 18015004

ミトコンドリアのリンパ器官への移植およびそのための組成物
Patent right, 菅沼 正司, 原島 秀吉, 山田 勇磨, 佐々木 大輔, 横田 貴史, ルカ・サイエンス株式会社
特願2023-220867, 27 Dec. 2023
特開2024-041813, 27 Mar. 2024
202403012899987552

サイズがより小さい単離されたミトコンドリアおよび単離されたミトコンドリアを内包している脂質膜ベースの小胞
Patent right, 菅沼 正司, 山田 勇磨, 原島 秀吉, 佐々木 大輔, 日比野 光恵, 太田 善浩, ルカ・サイエンス株式会社
特願2022-565028, 25 Dec. 2020
特表2023-508779, 03 Mar. 2023
202303002180247864

心不全の治療及び/又は予防に用いるための心筋幹細胞の製造方法
Patent right, 原島 秀吉, 山田 勇磨, 阿部 二郎, 武田 充人, ルカ・サイエンス株式会社
JP2017041250, 16 Nov. 2017
特許第7218897号, 30 Jan. 2023
202303007576008740

ミトコンドリアリボソームRNA変異検出用プライマーDNAセットを含むキット、ミトコンドリアリボソームRNA発現用核酸、前記核酸を封入してなる脂質膜構造体及びそれらの利用
Patent right, 山田 勇磨, 原島 秀吉, 丸山 美菜子, ルカ・サイエンス株式会社
特願2019-089566, 10 May 2019
特開2022-105226, 13 Jul. 2022
202203010989256665

リポソーム様ナノカプセル及びその製造方法
Patent right, 山田 勇磨, 原島 秀吉, 日比野 光恵, 佐藤 悠介, 渡慶次 学, 真栄城 正寿, ルカ・サイエンス株式会社
特願2019-070124, 01 Apr. 2019
特開2022-084962, 08 Jun. 2022
202203010499832250

ミトコンドリアDNAの3243位のアデニンからグアニンへの変異による変異tRNA Leuを検出する方法
Patent right, 山田 勇磨, 原島 秀吉, 国立大学法人北海道大学
特願2020-073420, 16 Apr. 2020
特開2021-168625, 28 Oct. 2021
202103000075425377

組換え発現ベクター及び当該ベクターを封入した脂質膜構造体
Patent right, 原島 秀吉, 山田 勇磨, 石川 卓哉, 秋田 英万, ルカ・サイエンス株式会社
特願2017-552752, 28 Nov. 2016
特許第6966755号, 26 Oct. 2021
202103004447787780

水溶性化合物を内包した脂質ナノ粒子
Patent right, 山田 勇磨, 原島 秀吉, 国立大学法人北海道大学
特願2020-071309, 10 Apr. 2020
特開2021-167289, 21 Oct. 2021
202103014885420380

ミトコンドリアのリンパ器官への移植およびそのための組成物
Patent right, 菅沼 正司, 原島 秀吉, 山田 勇磨, 佐々木 大輔, 横田 貴史, ルカ・サイエンス株式会社
JP2019036011, 13 Sep. 2019
WO2020-054829, 19 Mar. 2020
202103020804951832

組換え発現ベクター及び当該ベクターを封入した脂質膜構造体
Patent right, 原島 秀吉, 山田 勇磨, 石川 卓哉, 秋田 英万, 国立大学法人北海道大学
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WO2017-090763, 01 Jun. 2017
201803005641031779

機能性タンパク質を細胞内に送達するためのキャリア
Patent right, 原島 秀吉, 山田 勇磨, サンドラ, ミレーナ, ベルガラ ペレッツ, 国立大学法人北海道大学
特願2012-063815, 21 Mar. 2012
特開2015-110522, 18 Jun. 2015
201503019802774309

脂質膜構造体、脂質膜構造体の製造方法および1の目的物質を1枚の脂質膜で封入する方法
Patent right, 原島 秀吉, 山田 勇磨, 鈴木 亮佑, 国立大学法人北海道大学
JP2012054602, 24 Feb. 2012
WO2012-117971, 07 Sep. 2012
201403021176716730

目的物質をミトコンドリア内に送達可能な脂質膜構造体
Patent right, 山田 勇磨, 秋田 英万, 小暮 健太朗, 紙谷 浩之, 原島 秀吉, 菊池 寛, 小林 英夫, 国立大学法人北海道大学
特願2007-507194, 09 Mar. 2006
特許第5067733号
24 Aug. 2012
201303043484260566

核酸の放出性に優れたリポソームベクター
Patent right, 原島 秀吉, 山田 勇磨, 野村 卓, 由井 伸彦, 上遠野 亮, 山下 敦, 国立大学法人北海道大学, 国立大学法人北陸先端科学技術大学院大学
特願2008-305165, 28 Nov. 2008
特開2010-126505, 10 Jun. 2010
201003008997969186

一遺伝子ナノ粒子のパッケージング法
Patent right, 山田 勇磨, 鈴木 亮佑, 原島 秀吉, 国立大学法人 北海道大学
特願2008-068259, 17 Mar. 2008
特開2009-221165, 01 Oct. 2009
200903046276417326

目的物質をミトコンドリア内に送達可能な脂質膜構造体
Patent right, 山田 勇磨, 秋田 英万, 小暮 健太朗, 紙谷 浩之, 原島 秀吉, 菊池 寛, 小林 英夫, 国立大学法人 北海道大学, 第一三共株式会社
JP2006304656, 09 Mar. 2006
WO2006-095837, 14 Sep. 2006
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