Senso-immunology: the hidden relationship between sensory system and immune system.
Satoshi Miyamoto, Yasunori Takayama, Takeshi Kondo, Kenta Maruyama
Journal of bone and mineral metabolism, 2024年07月26日, ［国内誌］
英語, 研究論文（学術雑誌）, The primary sensory neurons involved in pain perception express various types of receptor-type ion channels at their nerve endings. These molecules are responsible for triggering neuronal excitation, translating environmental stimuli into pain signals. Recent studies have shown that acute nociception, induced by neuronal excitation, not only serves as a sensor for signaling life-threatening situations but also modulates our pathophysiological conditions. This modulation occurs through the release of neuropeptides by primary sensory neurons excited by nociceptive stimuli, which directly or indirectly affect peripheral systems, including immune function. Senso-immunology, an emerging research field, integrates interdisciplinary studies of pain and immunology and has garnered increasing attention in recent years. This review provides an overview of the systemic pathophysiological functions regulated by receptor-type ion channels, such as transient receptor potential (TRP) channels in primary sensory neurons, from the perspective of senso-immunology.

Neuroimmune modulation by tryptophan derivatives in neurological and inflammatory disorders.
Takeshi Kondo, Yuka Okada, Saika Shizuya, Naoko Yamaguchi, Shigetsugu Hatakeyama, Kenta Maruyama
European journal of cell biology, 103, 2, 151418, 151418, 2024年05月08日, ［筆頭著者］, ［国際誌］
英語, 研究論文（学術雑誌）, The nervous and immune systems are highly developed, and each performs specialized physiological functions. However, they work together, and their dysfunction is associated with various diseases. Specialized molecules, such as neurotransmitters, cytokines, and more general metabolites, are essential for the appropriate regulation of both systems. Tryptophan, an essential amino acid, is converted into functional molecules such as serotonin and kynurenine, both of which play important roles in the nervous and immune systems. The role of kynurenine metabolites in neurodegenerative and psychiatric diseases has recently received particular attention. Recently, we found that hyperactivity of the kynurenine pathway is a critical risk factor for septic shock. In this review, we first outline neuroimmune interactions and tryptophan derivatives and then summarized the changes in tryptophan metabolism in neurological disorders. Finally, we discuss the potential of tryptophan derivatives as therapeutic targets for neuroimmune disorders.

Sez6l2 autoimmunity in a large cohort study.
Megumi Abe, Hiroaki Yaguchi, Akihiko Kudo, Azusa Nagai, Shinichi Shirai, Ikuko Takahashi-Iwata, Masaaki Matsushima, Naoko Nakamura, Kenji Isahaya, Yoshihisa Yamano, Shinji Ashida, Takashi Kasai, Keiko Tanaka, Masashi Watanabe, Takeshi Kondo, Hidehisa Takahashi, Shigetsugu Hatakeyama, Akira Takekoshi, Akio Kimura, Takayoshi Shimohata, Ichiro Yabe
Journal of neurology, neurosurgery, and psychiatry, 94, 8, 667, 668, 2023年08月, ［国際誌］
英語

Lipid‐correlated alterations in the transcriptome are enriched in several specific pathways in the postmortem prefrontal cortex of Japanese patients with schizophrenia
Wataru Arihisa, Takeshi Kondo, Katsushi Yamaguchi, Junya Matsumoto, Hiroki Nakanishi, Yasuto Kunii, Hiroyasu Akatsu, Mizuki Hino, Yoshio Hashizume, Shumpei Sato, Shinji Sato, Shin‐Ichi Niwa, Hirooki Yabe, Takehiko Sasaki, Shuji Shigenobu, Mitsutoshi Setou
Neuropsychopharmacology Reports, Wiley, 2023年07月27日
研究論文（学術雑誌）, Abstract

Aims

Schizophrenia is a chronic relapsing psychiatric disorder that is characterized by many symptoms and has a high heritability. There were studies showing that the phospholipid abnormalities in subjects with schizophrenia (Front Biosci, S3, 2011, 153; Schizophr Bull, 48, 2022, 1125; Sci Rep, 7, 2017, 6; Anal Bioanal Chem, 400, 2011, 1933). Disturbances in prefrontal cortex phospholipid and fatty acid composition have been reported in subjects with schizophrenia (Sci Rep, 7, 2017, 6; Anal Bioanal Chem, 400, 2011, 1933; Schizophr Res, 215, 2020, 493; J Psychiatr Res, 47, 2013, 636; Int J Mol Sci, 22, 2021). For exploring the signaling pathways contributing to the lipid changes in previous study (Sci Rep, 7, 2017, 6), we performed two types of transcriptome analyses in subjects with schizophrenia: an unbiased transcriptome analysis solely based on RNA‐seq data and a correlation analysis between levels of gene expression and lipids.

Methods

RNA‐Seq analysis was performed in the postmortem prefrontal cortex from 10 subjects with schizophrenia and 5 controls. Correlation analysis between the transcriptome and lipidome from 9 subjects, which are the same samples in the previous lipidomics study (Sci Rep, 7, 2017, 6).

Results

Extraction of differentially expressed genes (DEGs) and further sequence and functional group analysis revealed changes in gene expression levels in phosphoinositide 3‐kinase (PI3K)‐Akt signaling and the complement system. In addition, a correlation analysis clarified alterations in ether lipid metabolism pathway, which is not found as DEGs in transcriptome analysis alone.

Conclusions

This study provided results of the integrated analysis of the schizophrenia‐associated transcriptome and lipidome within the PFC and revealed that lipid‐correlated alterations in the transcriptome are enriched in specific pathways including ether lipid metabolism pathway.

Senso-immunology: the past, present, and future.
Satoshi Miyamoto, Takeshi Kondo, Kenta Maruyama
Journal of biochemistry, 2023年07月18日, ［国際誌］
英語, 研究論文（学術雑誌）, Pain and mechanical stimulation are thought to be alarm systems that alert the brain to physical abnormalities. When we experience unpleasant feelings in infected or traumatized tissues, our awareness is directed to the afflicted region, prompting activities such as resting or licking the tissue. Despite extensive research into the molecular biology of nociceptors, it was unclear whether their role was limited to the generation and transmission of unpleasant feelings or whether they actively modulate the pathogenesis of infected or traumatized tissues. Recently, it has become clear how the sensory and immune systems interact with one another and share similar receptors and ligands to modify the pathogenesis of various diseases. In this paper, we summarize the mechanisms of crosstalk between the sensory and immune systems and the impact of this new interdisciplinary field, which should be dubbed "senso-immunology," on medical science.

TRIM22 negatively regulates MHC-II expression.
Ayano Inoue, Masashi Watanabe, Takeshi Kondo, Satoshi Hirano, Shigetsugu Hatakeyama
Biochimica et biophysica acta. Molecular cell research, 1869, 10, 119318, 119318, 2022年06月28日, ［国際誌］
英語, 研究論文（学術雑誌）, The development of cancer treatment has recently achieved a remarkable breakthrough, and checkpoint blockade immunotherapy has received much attention. To enhance the therapeutic efficacy of checkpoint blockade immunotherapy, recent studies have revealed the importance of activation of CD4+ T cells via an increase in major histocompatibility complex (MHC) class II molecules in cancer cells. Here, we demonstrate that tripartite motif-containing (TRIM) 22, negatively regulates MHC-II expression. Gene knockout of TRIM22 using Cas9-sgRNAs led to an increase of MHC-II proteins, while TRIM22 overexpression remarkably decreased MHC-II proteins. mRNA levels of MHC-II and class II transactivator (CIITA), which plays an essential role in the regulation of MHC-II transcription, were not affected by TRIM22. Furthermore, TRIM22 knockout did not suppress the degradation of MHC-II protein but rather promoted it. These results suggest that TRIM22 decreases MHC-II protein levels through a combination of multiple mechanisms other than transcription or degradation. We showed that inhibition of TRIM22 can increase the amount of MHC-II expression in cancer cells, suggesting a possibility of providing the biological basis for a possible therapeutic target to potentiate checkpoint blockade immunotherapy.

Lipid-correlated alterations in the transcriptome are enriched in several specific pathways in the postmortem prefrontal cortex of Japanese patients with schizophrenia
Wataru Arihisa, Takeshi Kondo, Katsushi Yamaguchi, Junya Matsumoto, Hiroki Nakanishi, Yasuto Kunii, Hiroyasu Akatsu, Mizuki Hino, Yoshio Hashizume, Shumpei Sato, Shinji Sato, Shin-Ichi Niwa, Hirooki Yabe, Takehiko Sasaki, Shuji Shigenobu, Mitsutoshi Setou
bioRxiv, Cold Spring Harbor Laboratory, 2022年03月16日
Background: Schizophrenia is a chronic relapsing psychiatric disorder that is characterized by many symptoms and has a high heritability. A previous study showed that specific lipid molecules belong to phosphatidylinositol (PI) and phosphatidylserine (PS) was reduced in the postmortem prefrontal cortex of patients with schizophrenia. However, signaling pathways contributing to the lipid changes remain unknown. Here we performed two types of transcriptome analyses in patients with schizophrenia: an un-biased transcriptome analysis solely based on RNA-seq data and a correlation analysis between levels of gene expression and lipids.Methodology/Principal Findings: RNA-Seq analysis was performed in the postmortem prefrontal cortex from 10 subjects with schizophrenia and 5 controls. Correlation analysis between the transcriptome and lipidome from 9 subjects which are the same samples in the previous lipidomics study (Table 1). Extraction of differentially expressed genes (DEGs) and further sequence and functional group analysis revealed changes of gene expression levels in phosphoinositide 3-kinase (PI3K)-Akt signaling and the complement system. In addition, a correlation analysis clarified alterations in several signaling/metabolic pathways including lipid-correlated genes, most of which are not found as DEGs in transcriptome analysis alone.Conclusions: This study provided results of the first integrated analysis of the schizophrenia-associated transcriptome and lipidome within the PFC and revealed that lipid-correlated alterations in the transcriptome are enriched in specific pathways including PI3K-Akt signaling.

Nociceptor-derived Reg3γ prevents endotoxic death by targeting kynurenine pathway in microglia
Erika Sugisawa, Takeshi Kondo, Yutaro Kumagai, Hiroki Kato, Yasunori Takayama, Kayako Isohashi, Eku Shimosegawa, Naoki Takemura, Yoshinori Hayashi, Takuya Sasaki, Mikaël M. Martino, Makoto Tominaga, Kenta Maruyama
Cell Reports, 38, 10, 110462, 110462, Elsevier BV, 2022年03月, ［査読有り］, ［筆頭著者］, ［国際誌］
英語, 研究論文（学術雑誌）, Nociceptors can fine-tune local or systemic immunity, but the mechanisms of nociceptive modulation in endotoxic death remain largely unknown. Here, we identified C-type lectin Reg3γ as a nociceptor-enriched hormone that protects the host from endotoxic death. During endotoxemia, nociceptor-derived Reg3γ penetrates the brain and suppresses the expression of microglial indoleamine dioxygenase 1, a critical enzyme of the kynurenine pathway, via the Extl3-Bcl10 axis. Endotoxin-administered nociceptor-null mice and nociceptor-specific Reg3γ-deficient mice exhibit a high mortality rate accompanied by decreased brain HK1 phosphorylation and ATP production despite normal peripheral inflammation. Such metabolic arrest is only observed in the brain, and aberrant production of brain quinolinic acid, a neurotoxic metabolite of the kynurenine pathway, causes HK1 suppression. Strikingly, the central administration of Reg3γ protects mice from endotoxic death by enhancing brain ATP production. By identifying nociceptor-derived Reg3γ as a microglia-targeted hormone, this study provides insights into the understanding of tolerance to endotoxic death.

Tubulin/microtubules as novel clozapine targets
Mizuki Hino, Takeshi Kondo, Yasuto Kunii, Junya Matsumoto, Akira Wada, Shin‐ichi Niwa, Mitsutoshi Setou, Hirooki Yabe
Neuropsychopharmacology Reports, 42, 1, 32, 41, Wiley, 2022年03月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, AIM: Clozapine is currently the only effective drug for treatment-resistant schizophrenia; nonetheless, its pharmacological mechanism remains unclear, and its administration is limited because of severe adverse effects. By comparing the binding proteins of clozapine and its derivative olanzapine, which is safer but less effective than clozapine, we attempted to clarify the mechanism of action specific to clozapine. METHODS: First, using the polyproline rod conjugates attached with clozapine or olanzapine, clozapine-binding proteins in extracts from the cerebra of 7-week-old ICR mice were isolated and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify proteins. Second, the effect of clozapine on tubulin polymerization was determined turbidimetrically. Finally, the cellular effects of clozapine were observed in HeLa cells by immunofluorescence microscopy. RESULTS: Alpha and β tubulins were the most abundant clozapine-binding proteins. We also found that clozapine directly binds with α and β tubulin heterodimers to inhibit their polymerization to form microtubules and disturbs the microtubule network, causing mitotic arrest in HeLa cells. CONCLUSION: These results suggest that α and β tubulin heterodimers are targeted by the clozapine and the microtubules are involved in the etiology of schizophrenia.

Fluticasone Propionate Suppresses Poly(I:C)-Induced ACE2 in Primary Human Nasal Epithelial Cells
Akira Nakazono, Yuji Nakamaru, Mahnaz Ramezanpour, Takeshi Kondo, Masashi Watanabe, Shigetsugu Hatakeyama, Shogo Kimura, Aya Honma, P. J. Wormald, Sarah Vreugde, Masanobu Suzuki, Akihiro Homma
Frontiers in Cellular and Infection Microbiology, 11, 655666, 655666, Frontiers Media SA, 2021年04月26日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, From the first detection in 2019, SARS-CoV-2 infections have spread rapidly worldwide and have been proven to cause an urgent and important health problem. SARS-CoV-2 cell entry depends on two proteins present on the surface of host cells, angiotensin-converting enzyme 2 (ACE2) and transmembrane protease serine 2 (TMPRSS2). The nasal cavity is thought to be one of the initial sites of infection and a possible reservoir for dissemination within and between individuals. However, it is not known how the expression of these genes is regulated in the nasal mucosa.

In this study, we examined whether the expression of ACE2 and TMPRSS2 is affected by innate immune signals in the nasal mucosa. We also investigated how fluticasone propionate (FP), a corticosteroid used as an intranasal steroid spray, affects the gene expression.

Primary human nasal epithelial cells (HNECs) were collected from the nasal mucosa and incubated with Toll-like receptor (TLR) agonists and/or fluticasone propionate (FP), followed by quantitative PCR, immunofluorescence, and immunoblot analyses.

Among the TLR agonists, the TLR3 agonist Poly(I:C) significantly increased ACE2 and TMPRSS2 mRNA expression in HNECs (ACE2 36.212±11.600-fold change, p<0.0001; TMPRSS2 5.598±2.434-fold change, p=0.031). The ACE2 protein level was also increased with Poly(I:C) stimulation (2.884±0.505-fold change, p=0.003). The Poly(I:C)-induced ACE2 expression was suppressed by co-incubation with FP (0.405±0.312-fold change, p=0.044).

The activation of innate immune signals via TLR3 promotes the expression of genes related to SARS-CoV2 cell entry in the nasal mucosa, although this expression is suppressed in the presence of FP. Further studies are required to evaluate whether FP suppresses SARS-CoV-2 viral cell entry.

A substrate-trapping strategy to find E3 ubiquitin ligase substrates identifies Parkin and TRIM28 targets
Masashi Watanabe, Yasushi Saeki, Hidehisa Takahashi, Fumiaki Ohtake, Yukiko Yoshida, Yusuke Kasuga, Takeshi Kondo, Hiroaki Yaguchi, Masanobu Suzuki, Hiroki Ishida, Keiji Tanaka, Shigetsugu Hatakeyama
Communications Biology, 3, 1, 592, 592, Springer Science and Business Media LLC, 2020年12月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）,
The identification of true substrates of an E3 ligase is biologically important but biochemically difficult. In recent years, several techniques for identifying substrates have been developed, but these approaches cannot exclude indirect ubiquitination or have other limitations. Here we develop an E3 ligase substrate-trapping strategy by fusing a tandem ubiquitin-binding entity (TUBE) with an anti-ubiquitin remnant antibody to effectively identify ubiquitinated substrates. We apply this method to one of the RBR-type ligases, Parkin, and to one of the RING-type ligases, TRIM28, and identify previously unknown substrates for TRIM28 including cyclin A2 and TFIIB. Furthermore, we find that TRIM28 promotes cyclin A2 ubiquitination and degradation at the G1/S phase and suppresses premature entry into S phase. Taken together, the results indicate that this method is a powerful tool for comprehensively identifying substrates of E3 ligases.

Zinc Finger Protein St18 Protects against Septic Death by Inhibiting VEGF-A from Macrophages
Kenta Maruyama, Hiroyasu Kidoya, Naoki Takemura, Erika Sugisawa, Osamu Takeuchi, Takeshi Kondo, Mohammed Mansour Abbas Eid, Hiroki Tanaka, Mikaël M. Martino, Nobuyuki Takakura, Yasunori Takayama, Shizuo Akira, Alexis Vandenbon, Yutaro Kumagai
Cell Reports, 32, 2, 107906, 107906, Elsevier BV, 2020年07月, ［査読有り］
研究論文（学術雑誌）

RNA Sensing by Gut Piezo1 Is Essential for Systemic Serotonin Synthesis
Erika Sugisawa, Yasunori Takayama, Naoki Takemura, Takeshi Kondo, Shigetsugu Hatakeyama, Yutaro Kumagai, Masataka Sunagawa, Makoto Tominaga, Kenta Maruyama
Cell, 182, 3, 609, 624.e21, Elsevier BV, 2020年07月, ［査読有り］, ［筆頭著者］
研究論文（学術雑誌）

Preferential Incorporation of Administered Eicosapentaenoic Acid Into Thin-Cap Atherosclerotic Plaques.
Tomohito Sato, Makoto Horikawa, Shiro Takei, Fumiyoshi Yamazaki, Takashi K Ito, Takeshi Kondo, Takanobu Sakurai, Tomoaki Kahyo, Koji Ikegami, Shumpei Sato, Ryota Sato, Yasutaka Jinno, Hiroyuki Kawano, Satoko Naoe, Makoto Arita, Yukiyasu Kashiwagi, Mitsutoshi Setou
Arteriosclerosis, thrombosis, and vascular biology, 39, 9, 1802, 1816, 2019年09月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, OBJECTIVE: n-3 polyunsaturated fatty acids, especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), have beneficial effects on atherosclerosis. Although specific salutary actions have been reported, the detailed distribution of n-3 polyunsaturated fatty acids in plaque and their relevance in disease progression are unclear. Our aim was to assess the pharmacodynamics of EPA and DHA and their metabolites in atherosclerotic plaques. Approach and Results: Apolipoprotein E-deficient (Apoe-/-) mice were fed a Western diet supplemented with EPA (1%, w/w) or DHA (1%, w/w) for 3 weeks. Imaging mass spectrometry analyses were performed in the aortic root and arch of the Apoe-/- mice to evaluate the distribution of EPA, DHA, their metabolites and the lipids containing EPA or DHA in the plaques. Liquid chromatography-mass spectrometry and histological analysis were also performed. The intima-media thickness of atherosclerotic plaque decreased in plaques containing free EPA and EPAs attached with several lipids. EPA was distributed more densely in the thin-cap plaques than in the thick-cap plaques, while DHA was more evenly distributed. In the aortic root, the distribution of total EPA level and cholesteryl esters containing EPA followed a concentration gradient from the vascular endothelium to the media. In the aortic arch, free EPA and 12-hydroxy-EPA colocalized with M2 macrophage. CONCLUSIONS: Administered EPA tends to be incorporated from the vascular lumen side and preferentially taken into the thin-cap plaque.

The ATP Transporter VNUT Mediates Induction of Dectin-1-Triggered Candida Nociception.
Kenta Maruyama, Yasunori Takayama, Erika Sugisawa, Yu Yamanoi, Takashi Yokawa, Takeshi Kondo, Ken-Ichi Ishibashi, Bikash Ranjan Sahoo, Naoki Takemura, Yuki Mori, Hisashi Kanemaru, Yutaro Kumagai, Mikaël M Martino, Yoshichika Yoshioka, Hisao Nishijo, Hiroki Tanaka, Atsushi Sasaki, Naohito Ohno, Yoichiro Iwakura, Yoshinori Moriyama, Masatoshi Nomura, Shizuo Akira, Makoto Tominaga
iScience, 6, 306, 318, 2018年08月31日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Candida albicans infection can cause skin, vulvar, or oral pain. Despite the obvious algesic activity of C. albicans, the molecular mechanisms of fungal nociception remain largely unknown. Here we show that the C. albicans-specific signaling pathway led to severe mechanical allodynia. We discovered that C. albicans-derived β-glucan stimulated nociceptors depending on Dectin-1, and two pathways in inflammatory pain. The major pathway operates via the Dectin-1-mediated ATP-P2X3/P2X2/3 axis through intercellular relationships between keratinocytes and primary sensory neurons, which depends on the ATP transporter vesicular nucleotide transporter (VNUT). The other pathway operates via the Dectin-1-mediated PLC-TRPV1/TRPA1 axis in primary sensory neurons. Intriguingly, C. albicans-derived β-glucan has the ability to enhance histamine-independent pruritus, and VNUT inhibitor clodronate can be used to treat unpleasant feelings induced by β-glucan. Collectively, this is the first report to indicate that Dectin-1 and VNUT mediated innate sensory mechanisms that detect fungal infection.

A power law distribution of metabolite abundance levels in mice regardless of the time and spatial scale of analysis.
Shumpei Sato, Makoto Horikawa, Takeshi Kondo, Tomohito Sato, Mitsutoshi Setou
Scientific reports, 8, 1, 10315, 10315, 2018年07月09日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Biomolecule abundance levels change with the environment and enable a living system to adapt to the new conditions. Although, the living system maintains at least some characteristics, e.g. homeostasis. One of the characteristics maintained by a living system is a power law distribution of biomolecule abundance levels. Previous studies have pointed to a universal characteristic of biochemical reaction networks, with data obtained from lysates of multiple cells. As a result, the spatial scale of the data related to the power law distribution of biomolecule abundance levels is not clear. In this study, we researched the scaling law of metabolites in mouse tissue with a spatial scale of quantification that was changed stepwise between a whole-tissue section and a single-point analysis (25 μm). As a result, metabolites in mouse tissues were found to follow the power law distribution independently of the spatial scale of analysis. Additionally, we tested the temporal changes by comparing data from younger and older mice. Both followed similar power law distributions, indicating that metabolite composition is not diversified by aging to disrupt the power law distribution. The power law distribution of metabolite abundance is thus a robust characteristic of a living system regardless of time and space.

Netrins as prophylactic targets in skeletal diseases: A double-edged sword?
Kenta Maruyama, Naoki Takemura, Mikael M. Martino, Takeshi Kondo, Shizuo Akira
PHARMACOLOGICAL RESEARCH, 122, 46, 52, ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD, 2017年08月, ［査読有り］
英語, 研究論文（学術雑誌）, The netrin family of proteins are involved in axon guidance during central nervous system development. In vertebrates, two membrane bound forms and five secreted forms of netrin have been reported. In addition to their critical role in neural morphogenesis, a growing number of reports suggest that netrin family proteins also play a role in inflammatory conditions, angiogenesis, and turnorigenesis. In these processes, Unc5 and DCC family proteins serve as receptors of netrin proteins. Recently, it was reported that some netrin family proteins may be involved in the pathogenesis of skeletal diseases including osteoporosis and arthritis. For example, administration of secreted netrin family proteins such as netrin 1 and netrin 4 has prophylactic potential in pathogenic bone degradation in mice. However, netrin 1 blocking antibody also protects mice from inflammatory bone destruction. Therefore, netrin family proteins are involved in the regulation of bone homeostasis, but their bona fide roles in the skeletal system remain controversial. In this review, we discuss the osteo-innate-immune functions of the netrin family of proteins, and summarize their therapeutic potential. (C) 2017 Elsevier Ltd. All rights reserved.

Nociceptors Boost the Resolution of Fungal Osteoinflammation via the TRP Channel-CGRP-Jdp2 Axis
Kenta Maruyama, Yasunori Takayama, Takeshi Kondo, Ken-ichi Ishibashi, Bikash Ranjan Sahoo, Hisashi Kanemaru, Yutaro Kumagai, Mikael M. Martino, Hiroki Tanaka, Naohito Ohno, Yoichiro Iwakura, Naoki Takemura, Makoto Tominaga, Shizuo Akira
CELL REPORTS, 19, 13, 2730, 2742, CELL PRESS, 2017年06月, ［査読有り］
英語, 研究論文（学術雑誌）, Candida albicans can enter skeletal tissue through a skin wound in an immunocompromised host or by contamination during orthopedic surgery. Such Candida osteomyelitis is accompanied by severe pain and bone destruction. It is established that nociceptor innervation occurs in skin and bone, but the mechanisms of nociceptive modulation in fungal inflammation remain unclear. In this study, we show that C. albicans stimulates Nav1.8-positive nociceptors via the beta-glucan receptor Dectin-1 to induce calcitonin gene-related peptide (CGRP). This induction of CGRP is independent of Bcl-10 or Malt-1 but dependent on transient receptor potential cation channel subfamily V member 1 (TRPV1)/transient receptor potential cation channel subfamily A member 1 (TRPA1) ion channels. Hindpaw beta-glucan injection after av1.8-positive nociceptor ablation or in TRPV1/TRPA1 deficiency showed dramatically increased osteoinflammation accompanied by impaired CGRP production. Strikingly, CGRP suppressed beta-glucan-induced inflammation and osteoclast multinucleation via direct suppression of nuclear factor-kappa B (NF-kappa B) p65 by the transcriptional repressor Jdp2 and inhibition of actin polymerization, respectively. These findings clearly suggest a role for Dectin-1-mediated sensocrine pathways in the resolution of fungal osteoinflammation.

Downregulation of GNA13-ERK network in prefrontal cortex of schizophrenia brain identified by combined focused and targeted quantitative proteomics
Mio Hirayama-Kurogi, Yohei Takizawa, Yasuto Kunii, Junya Matsumoto, Akira Wada, Mizuki Hino, Hiroyasu Akatsu, Yoshio Hashizume, Sakon Yamamoto, Takeshi Kondo, Shingo Ito, Masanori Tachikawa, Shin-Ichi Niwa, Hirooki Yabe, Tetsuya Terasaki, Mitsutoshi Setou, Sumio Ohtsuki
JOURNAL OF PROTEOMICS, 158, 31, 42, ELSEVIER SCIENCE BV, 2017年03月, ［査読有り］
英語, 研究論文（学術雑誌）, Schizophrenia is a disabling mental illness associated with dysfunction of the prefrontal cortex, which affects cognition and emotion.The purpose of the present study was to identify altered molecular networks in the prefrontal cortex of schizophrenia patients by comparing protein expression levels in autopsied brains of patients and controls, using a combination of targeted and focused quantitative proteomics. We selected 125 molecules possibly related to schizophrenia for quantification by knowledge-based targeted proteomics. Among the quantified molecules, GRIK4 and MAO-B were significantly decreased in plasma membrane and cytosolic fractions, respectively, of prefrontal cortex. Focused quantitative proteomics identified 15 increased and 39 decreased proteins. Network analysis identified "GNA13-ERK1-elF4G2 signaling" as a downregulated network, and proteins involved in this network were significantly decreased. Furthermore, searching downstream of eIF4G2 revealed that elF4A1/2 and CYFIP1 were decreased, suggesting that downregulation of the network suppresses expression of CYFIP1, which regulates actin remodeling and is involved in axon outgrowth and spine formation. Downregulation of this signaling seems likely to impair axon formation and synapse plasticity of neuronal cells, and could be associated with development of cognitive impairment in the pathology of schizophrenia.
Biological significance: The present study compared the proteome of the prefrontal cortex between schizophrenia patients and healthy controls by means of targeted proteomics and global quantitative proteomics. Targeted proteomics revealed that GRIK4 and MAOB were significantly decreased among 125 putatively schizophrenia-related proteins in prefrontal cortex of schizophrenia patients. Global quantitative proteomics identified 54 differentially expressed proteins in schizophrenia brains. The protein profile indicates attenuation of "GNA13ERK signaling" in schizophrenia brain. In particular, EIF4G2 and CYFIP1, which are located downstream of theGNA13-ERK network, were decreased, suggesting that the attenuation of this signal network may cause impairment of axon formation and synapse plasticity in the brain of schizophrenia patients. Our results provide a novel insight into schizophrenia pathology, and could be helpful for drug development. (C) 2017 Elsevier B.V. All rights reserved.

Decreased 16:0/20:4-phosphatidylinositol level in the post-mortem prefrontal cortex of elderly patients with schizophrenia
Junya Matsumoto, Hiroki Nakanishi, Yasuto Kunii, Yuki Sugiura, Dai Yuki, Akira Wada, Mizuki Hino, Shin-Ichi Niwa, Takeshi Kondo, Michihiko Waki, Takahiro Hayasaka, Noritaka Masaki, Hiroyasu Akatsu, Yoshio Hashizume, Sakon Yamamoto, Shinji Sato, Takehiko Sasaki, Mitsutoshi Setou, Hirooki Yabe
SCIENTIFIC REPORTS, 7, 45050, NATURE PUBLISHING GROUP, 2017年03月, ［査読有り］
英語, 研究論文（学術雑誌）, The etiology of schizophrenia includes phospholipid abnormalities. Phospholipids are bioactive substances essential for brain function. To analyze differences in the quantity and types of phospholipids present in the brain tissue of patients with schizophrenia, we performed a global analysis of phospholipids in multiple brain samples using liquid chromatography electrospray ionization mass/mass spectrometry (LC-ESI/MS/MS) and imaging mass spectrometry (IMS). We found significantly decreased 16:0/20:4-phosphatidylinositol ( PI) levels in the prefrontal cortex (PFC) in the brains from patients with schizophrenia in the LC-ESI/MS/MS, and that the 16:0/20: 4-PI in grey matter was most prominently diminished according to the IMS experiments. Previous reports investigating PI pathology of schizophrenia did not identify differences in the sn-1 and sn-2 fatty acyl chains. This study is the first to clear the fatty acid composition of PI in brains from patients with schizophrenia. Alteration in the characteristic fatty acid composition of PI may also affect neuronal function, and could play a role in the etiology of schizophrenia. Although further studies are necessary to understand the role of reduced

Bone-protective Functions of Netrin 1 Protein
Kenta Maruyama, Takahiko Kawasaki, Masahide Hamaguchi, Motomu Hashimoto, Moritoshi Furu, Hiromu Ito, Takao Fujii, Naoki Takemura, Thangaraj Karuppuchamy, Takeshi Kondo, Takumi Kawasaki, Masahiro Fukasaka, Takuma Misawa, Tatsuya Saitoh, Yutaka Suzuki, Mikael M. Martino, Yutaro Kumagai, Shizuo Akira
JOURNAL OF BIOLOGICAL CHEMISTRY, 291, 46, 23854, 23868, AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, 2016年11月, ［査読有り］
英語, 研究論文（学術雑誌）, Netrin 1 was initially identified as an axon guidance factor, and recent studies indicate that it inhibits chemokine-directed monocyte migration. Despite its importance as a neuroimmune guidance cue, the role of netrin 1 in osteoclasts is largely unknown. Here we detected high netrin 1 levels in the synovial fluid of rheumatoid arthritis patients. Netrin 1 is potently expressed in osteoblasts and synovial fibroblasts, and IL-17 robustly enhances netrin 1 expression in these cells. The binding of netrin 1 to its receptor UNC5b on osteoclasts resulted in activation of SHP1, which inhibited VAV3 phosphorylation and RAC1 activation. This significantly impaired the actin polymerization and fusion, but not the differentiation of osteoclast. Strikingly, netrin 1 treatment prevented bone erosion in an autoimmune arthritis model and age-related bone destruction. Therefore, the netrin 1-UNC5b axis is a novel therapeutic target for bone-destructive diseases.

5-Azacytidine-induced Protein 2 (AZI2) Regulates Bone Mass by Fine-tuning Osteoclast Survival
Kenta Maruyama, Masahiro Fukasaka, Satoshi Uematsu, Osamu Takeuchi, Takeshi Kondo, Tatsuya Saitoh, Mikael M. Martino, Shizuo Akira
JOURNAL OF BIOLOGICAL CHEMISTRY, 290, 15, 9377, 9386, AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, 2015年04月, ［査読有り］
英語, 研究論文（学術雑誌）, 5-Azacytidine-induced protein 2 (AZI2) is a TNF receptor (TNFR)-associated factor family member-associated NF-kappa B activator-binding kinase 1-binding protein that regulates the production of IFNs. A previous in vitro study showed that AZI2 is involved in dendritic cell differentiation. However, the roles of AZI2 in immunity and its pleiotropic functions are unknown in vivo. Here we report that AZI2 knock-out mice exhibit normal dendritic cell differentiation in vivo. However, we found that adult AZI2 knock-out mice have severe osteoporosis due to increased osteoclast longevity. We revealed that the higher longevity of AZI2-deficient osteodasts is due to an augmented activation of proto-oncogene tyrosine-protein kinase Src (c-Src), which is a critical player in osteoclast survival. We found that AZI2 inhibits c-Src activity by regulating the activation of heat shock protein 90 (Hsp90), a chaperone involved in c-Src dephosphorylation. Furthermore, we demonstrated that AZI2 indirectly inhibits c-Src by interacting with the Hsp90 co-chaperone Cdc37. Strikingly, administration of a c-Src inhibitor markedly prevented bone loss in AZI2 knock-out mice. Together, these findings indicate that AZI2 regulates bone mass by fine-tuning osteoclast survival.

Single-cell time-of-flight secondary ion mass spectrometry reveals that human breast cancer stem cells have significantly lower content of palmitoleic acid compared to their counterpart non-stem cancer cells
Michihiko Waki, Yoshimi Ide, Itsuko Ishizaki, Yasuyuki Nagata, Noritaka Masaki, Eiji Sugiyama, Nobuya Kurabe, Dan Nicolaescu, Fumiyoshi Yamazaki, Takahiro Hayasaka, Koji Ikegami, Takeshi Kondo, Kiyoshi Shibata, Takanori Hiraide, Yumiko Taki, Hiroyuki Ogura, Norihiko Shiiya, Noriaki Sanada, Mitsutoshi Setou
BIOCHIMIE, 107, 73, 77, ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER, 2014年12月, ［査読有り］
英語, 研究論文（学術雑誌）, Lipids comprise the primary component of cell membranes. Imaging mass spectrometry is increasingly being used to visualize membranous lipids in clinical specimens, and it has revealed that abnormal lipid metabolism is related to the development of diseases. To characterize cell populations which are rare and sparsely localized in tissues, we conducted time-of-flight secondary ion mass spectrometry (TOF-SIMS) analyses of individual cells sorted by fluorescence activated cell sorting (FACS) and applied the method to analyze breast cancer stem cells (CSCs). TOF-SIMS analyses visualized phosphoric acids and four fatty acid (FA) species in the sorted CD45(-)/CD44(+)/CD24(-) CSCs, and these ions are suspected to have originated from membranous phospholipids as they were uniformly detected from the locus where the cells attached. Integrated ion intensity of palmitoleic acids [FA(16:1)] normalized by phosphoric acid signals were decreased significantly in CSCs as compared to that of CD45(-)/CD44(-)/CD24(+) non-stem cancer cells (NSCCs). This finding was supported by liquid chromatography coupled electrospray ionization-tandem mass spectrometry analysis, which revealed phosphatidylcholine (PC)(16:0/16:1) to be less abundant and PC(16:0/16:0) to be more abundant in CSCs as compared to NSCCs. Therefore, our novel method successfully provided lipid composition analysis of individual cells classified by the expression of a complex combination of cell-surface markers. The lipid compositions of CSCs originating from the heterogeneous cellular populations of clinical specimens were successfully characterized by this method. (C) 2014 Elsevier B.V. and Societe francaise de biochimie et biologie Moleculaire (SFBBM). All rights reserved.

Single cell lipidomics of SKBR-3 breast cancer cells by using time-of-flight secondary-ion mass spectrometry
Yoshimi Ide, Michihiko Waki, Itsuko Ishizaki, Yasuyuki Nagata, Fumiyoshi Yamazaki, Takahiro Hayasaka, Noritaka Masaki, Koji Ikegami, Takeshi Kondo, Kiyoshi Shibata, Hiroyuki Ogura, Noriaki Sanada, Mitsutoshi Setou
SURFACE AND INTERFACE ANALYSIS, 46, S1, 181, 184, WILEY-BLACKWELL, 2014年11月, ［査読有り］
英語, 研究論文（国際会議プロシーディングス）, Breast cancer is the most common cancer among women worldwide. The molecular characterization of breast tumor cells by using single-cell lipidomics remains relatively unexplored. Here, we introduce a time-of-flight secondary-ion mass spectrometry (TOF-SIMS) approach to visualize the lipids in individual breast cancer cells. The SKBR-3 breast cancer cell line was cultured and dispersed into individual cells. After attachment to a substrate, the cells were rinsed with ammonium acetate and were analyzed using TOF-SIMS. The instrument was operated with Bi-3(2+) as the primary ion. The distributions of ions, including positively charged phosphocholine, and negatively charged phosphates and fatty acids, were simultaneously visualized. These ions were distributed predominantly at the cell attachment sites. The signal intensities of fatty acid ions were determined from the mass spectra at the regions-of-interest. The results of fatty acid analyses on breast cancer cells were consistent with those of our previous study in which prominent expression of stearoyl-CoA desaturase 1 in breast cancer cells was demonstrated. Static TOF-SIMS was shown to be an effective method for determining the lipid molecular signature of the plasma membrane of individual breast cancer cells. Copyright (c) 2014 John Wiley & Sons, Ltd.

Nanoparticle-assisted laser desorption/ionization for metabolite imaging
Michihiko Waki, Eiji Sugiyama, Takeshi Kondo, Keigo Sano, Mitsutoshi Setou
Mass Spectrometry Imaging of Small Molecules, 1203, 159, 173, Springer Fachmedien, 2014年10月31日, ［査読有り］
英語, 論文集(書籍)内論文, Matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) has enabled the spatial analysis of various molecules, including peptides, nucleic acids, lipids, and drug molecules. To expand the capabilities of MALDI-IMS, we have established an imaging technique using metal nanoparticles (NPs) to visualize metabolites, termed nanoparticle-assisted laser desorption/ionization imaging mass spectrometry (nano-PALDI-IMS). By utilizing Ag-, Fe-, Au-, and TiO2 -derived NPs, we have succeeded in visualizing various metabolites, including fatty acid and glycosphingolipids, with higher sensitivity and spatial resolution than conventional techniques. Herein, we describe the practical experimental procedures and methods associated with nano-PALDI-IMS for the visualization of these molecules.

Strawberry notch homologue 2 regulates osteoclast fusion by enhancing the expression of DC-STAMP
Kenta Maruyama, Satoshi Uematsu, Takeshi Kondo, Osamu Takeuchi, Mikael M. Martino, Takumi Kawasaki, Shizuo Akira
JOURNAL OF EXPERIMENTAL MEDICINE, 210, 10, 1947, 1960, ROCKEFELLER UNIV PRESS, 2013年09月, ［査読有り］
英語, 研究論文（学術雑誌）, Osteoclasts are multinucleated cells formed by fusion of mononuclear precursors in response to receptor activator of nuclear factor kappa B (NF-kappa B) ligand (RANKL). We found that RANKL induced expression of the DExD/H helicase family corepressor strawberry notch homologue 2 (Sbno2). Previous in vitro studies showed that Sbno2 is induced by IL-10 and is involved in NF-kappa B repression. However, the role of Sbno2 in vivo and its pleiotropic functions are unknown. Sbno2 gene targeting resulted in normal NF-kappa B activation by TLR ligands. However, Sbno2-deficient mice exhibited increased bone mass due to impaired osteoclast fusion. Expression of dendritic cell-specific transmembrane protein (DC-STAMP), a critical player in osteoclast fusion, was significantly attenuated, and cell fusion of Sbno2-deficient osteoclasts was rescued by DC-STAMP. Sbno2 directly bound to T cell acute lymphocytic leukemia 1 (Tal1) and attenuated its inhibition of DC-STAMP expression, leading to activation of the DC-STAMP promoter by microphthalmia-associated transcription factor (MITF). Thus, Sbno2 plays a pivotal role in bone homeostasis in vivo by fine-tuning osteoclast fusion.

DNA damage sensor MRE11 recognizes cytosolic double-stranded DNA and induces type I interferon by regulating STING trafficking
Takeshi Kondo, Junya Kobayashi, Tatsuya Saitoh, Kenta Maruyama, Ken J. Ishii, Glen N. Barber, Kenshi Komatsu, Shizuo Akira, Taro Kawai
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 110, 8, 2969, 2974, NATL ACAD SCIENCES, 2013年02月, ［査読有り］, ［筆頭著者］
英語, 研究論文（学術雑誌）, Double-stranded DNA (dsDNA) derived from pathogen-or host-damaged cells triggers innate immune responses when exposed to cytoplasm. However, the machinery underlying the primary recognition of intracellular dsDNA is obscure. Here we show that the DNA damage sensor, meiotic recombination 11 homolog A (MRE11), serves as a cytosolic sensor for dsDNA. Cells with a mutation of MRE11 gene derived from a patient with ataxia-telangiectasia-like disorder, and cells in which Mre11 was knocked down, had defects in dsDNA-induced type I IFN production. MRE11 physically interacted with dsDNA in the cytoplasm and was required for activation of stimulator of IFN genes (STING) and IRF3. RAD50, a binding protein to MRE11, was also required for dsDNA responses, whereas NBS1, another binding protein to MRE11, was dispensable. Collectively, our results suggest that the MRE11-RAD50 complex plays important roles in recognition of dsDNA and initiation of STING-dependent signaling, in addition to its role in DNA-damage responses.

The Transcription Factor Jdp2 Controls Bone Homeostasis and Antibacterial Immunity by Regulating Osteoclast and Neutrophil Differentiation
Kenta Maruyama, Masahiro Fukasaka, Alexis Vandenbon, Tatsuya Saitoh, Takumi Kawasaki, Takeshi Kondo, Kazunari K. Yokoyama, Hiroyasu Kidoya, Nobuyuki Takakura, Daron Standley, Osamu Takeuchi, Shizuo Akira
IMMUNITY, 37, 6, 1024, 1036, CELL PRESS, 2012年12月, ［査読有り］
英語, 研究論文（学術雑誌）, Jdp2 is an AP-1 family transcription factor that regulates the epigenetic status of histones. Previous in vitro studies revealed that Jdp2 is involved in osteoclastogenesis. However, the roles of Jdp2 in vivo and its pleiotropic functions are largely unknown. Here we generated Jdp2(-/-) mice and discovered its crucial roles not only in bone metabolism but also in differentiation of neutrophils. Jdp2(-/-) mice exhibited osteopetrosis resulting from impaired osteoclastogenesis. Jdp2(-/-) neutrophils were morphologically normal but had impaired surface expression of Ly6G, bactericidal function, and apoptosis. We also found that ATF3 was an inhibitor of neutrophil differentiation and that Jdp2 directly suppresses its expression via inhibition of histone acetylation. Strikingly, Jdp2(-/-) mice were highly susceptible to Staphylococcus aureus and Candida albicans infection. Thus, Jdp2 plays pivotal roles in in vivo bone homeostasis and host defense by regulating osteoclast and neutrophil differentiation.

Dissecting negative regulation of Toll-like receptor signaling
Takeshi Kondo, Taro Kawai, Shizuo Akira
TRENDS IN IMMUNOLOGY, 33, 9, 449, 458, ELSEVIER SCI LTD, 2012年09月, ［査読有り］, ［筆頭著者］
英語, 研究論文（学術雑誌）, Toll-like receptors (TLRs) sense invading microbial pathogens and play crucial roles in the activation of innate and adaptive immunity. However, excessive TLR activation can disrupt immune homeostasis, and may be responsible for the development of autoimmune and inflammatory diseases. As such, the molecules and pathways that negatively control TLR signaling have been intensively investigated. Here, we discuss recent insights into the negative regulation of TLR signaling, with focus on three major mechanisms: (i) dissociation of adaptor complexes; (ii) degradation of signal proteins; and WO transcriptional regulation. We also highlight how pathogens negatively target TLR signaling as a strategy to evade the host immune response.

TRAF family member-associated NF-κB activator (TANK) is a negative regulator of osteoclastogenesis and bone formation.
Maruyama K, Kawagoe T, Kondo T, Akira S, Takeuchi O
J Biol Chem., 287, 34, 29114, 29124, AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, 2012年08月, ［査読有り］
英語, 研究論文（学術雑誌）, The differentiation of bone-resorbing osteoclasts is induced by RANKL signaling, and leads to the activation of NF-kappa B via TRAF6 activation. TRAF family member-associated NF-kappa B activator (TANK) acts as a negative regulator of Toll-like receptors (TLRs) and B-cell receptor (BCR) signaling by inhibiting TRAF6 activation. Tank(-/-) mice spontaneously develop autoimmune glomerular nephritis in an IL-6-dependent manner. Despite its importance in the TCRs and BCR-activated TRAF6 inhibition, the involvement of TANK in RANKL signaling is poorly understood. Here, we report that TANK is a negative regulator of osteoclast differentiation. The expression levels of TANK mRNA and protein were up-regulated during RANKL-induced osteoclastogenesis, and overexpression of TANK in vitro led to a decrease in osteoclast formation. The in vitro osteoclastogenesis of Tank(-/-) cells was significantly increased, accompanied by increased ubiquitination of TRAF6 and enhanced canonical NF-kappa B activation in response to RANKL stimulation. Tank(-/-) mice showed severe trabecular bone loss, but increased cortical bone mineral density, because of enhanced bone erosion and formation. TANK mRNA expression was induced during osteoblast differentiation and Tank(-/-) osteoblasts exhibited enhaced NF-kappa B activation, IL-11 expression, and bone nodule formation than wild-type control cells. Finally, wild-type mice transplanted with bone marrow cells from Tank(-/-) mice showed trabecular bone loss analogous to that in Tank(-/-) mice. These findings demonstrate that TANK is critical for osteoclastogenesis by regulating NF-kappa B, and is also important for proper bone remodeling.

TRIM59 interacts with ECSIT and negatively regulates NF-κB and IRF-3/7-mediated signal pathways.
Kondo T, Watanabe M, Hatakeyama S
Biochem Biophys Res Commun., 422, 3, 501, 507, Elsevier, 2012年06月, ［査読有り］, ［筆頭著者］
英語, 研究論文（学術雑誌）, Innate immune responses are triggered by pathogen-associated molecular patterns (PAMPs) through pattern recognition receptors (PRRs) and then activate intracellular signaling pathways including NF-κB and interferon regulatory factors. Recently, it has been reported that tripartite motif (TRIM) proteins function as crucial regulators via ubiquitin-mediated modifications for these signaling pathways. In this study, we showed that one of the TRIM family ubiquitin ligases, TRIM59, interacts with ECSIT as an adaptor protein required for the TLR-mediated transduction pathway. Luciferase reporter assays using reporter plasmids including NF-κB responsive element, interferon β (IFN-β) promoter and interferon-sensitive response element (ISRE) showed that overexpression of TRIM59 repressed their transcriptional activities, whereas knockdown of TRIM59 enhanced their transcriptional activities. Furthermore, TRIM59 inhibited phosphorylation and dimerization of IRF3 and IRF7, suggesting that TRIM59 negatively regulates upstream kinases for IRFs. These findings indicate that TRIM59 may serve as a multifunctional regulator for innate immune signaling pathways.

Plasma gelsolin facilitates interaction between β2 glycoprotein I and α5β1 integrin.
Bohgaki M, Matsumoto M, Atsumi T, Kondo T, Yasuda S, Horita T, Nakayama KI, Okumura F, Hatakeyama S, Koike T
J Cell Mol Med., 15, 1, 141, 151, WILEY-BLACKWELL, 2011年01月, ［査読有り］
英語, 研究論文（学術雑誌）, Antiphospholipid syndrome (APS) is characterized by thrombosis and the presence of antiphospholipid antibodies (aPL) that directly recognizes plasma beta(2)-glycoprotein I (beta(2)GPI). Tissue factor (TF), the major initiator of the extrinsic coagulation system, is induced on monocytes by aPL in vitro, explaining in part the pathophysiology in APS. We previously reported that the mitogen-activated protein kinase (MAPK) pathway plays an important role in aPL-induced TF expression on monocytes. In this study, we identified plasma gelsolin as a protein associated with beta(2)GPI by using immunoaffinity chromatography and mass spectrometric analysis. An in vivo binding assay showed that endogenous beta(2)GPI interacts with plasma gelsolin, which binds to integrin a(5)beta(1) through fibronectin. The tethering of beta(2)GPI to monoclonal anti-beta(2)GPI autoantibody on the cell surface was enhanced in the presence of plasma gelsolin. Immunoblot analysis demonstrated that p38 MAPK protein was phosphorylated by monoclonal anti-beta(2)GPI antibody treatment, and its phosphorylation was attenuated in the presence of anti-integrin a(5)beta(1) antibody. Furthermore, focal adhesion kinase, a downstream molecule of the fibronectin-integrin signalling pathway, was phosphorylated by anti-beta(2)GPI antibody treatment. These results indicate that molecules including gelsolin and integrin are involved in the anti-beta(2)GPI antibody-induced MAPK pathway on monocytes and that integrin is a possible therapeutic target to modify a prothrombotic state in patients with APS.

鼻副鼻腔粘膜悪性黒色腫におけるTRIM27の検討　　　　　　　　　　　　　　　
木村 将吾, 中丸 裕爾, 鈴木 正宣, 中薗 彬, 本間 あや, 渡邉 良亮, 加納 里志, 対馬 那由多, 鈴木 崇祥, 井戸川 寛志, 本間 明宏, 渡部 昌, 近藤 豪, 畠山 鎮次, 日本鼻科学会会誌, 63, 3, 336, 336, 2024年09月
(一社)日本鼻科学会, 日本語

乳癌細胞株におけるubiqultin-like 3(UBL3)の増殖・浸潤への寄与検証　　　　　　　　　　　　　　　
高塚 大輝, 高梨 裕典, 淺野 祐子, 綿引 麻耶, 山崎 宏和, 小泉 圭, 華表 友暁, 瀬藤 光利, 近藤 豪, 椎谷 紀彦, 日本乳癌学会総会プログラム抄録集, 31回, 232, 232, 2023年06月
(一社)日本乳癌学会, 日本語

乳癌細胞株におけるubiqultin-like 3(UBL3)の増殖・浸潤への寄与検証　　　　　　　　　　　　　　　
高塚 大輝, 高梨 裕典, 淺野 祐子, 綿引 麻耶, 山崎 宏和, 小泉 圭, 華表 友暁, 瀬藤 光利, 近藤 豪, 椎谷 紀彦, 日本乳癌学会総会プログラム抄録集, 31回, 232, 232, 2023年06月
(一社)日本乳癌学会, 日本語

効率的な遺伝子発現制御を可能にするCRISPRi法の開発　　　　　　　　　　　　　　　
大木 のどか, 近藤 豪, 畠山 鎮次, 日本生化学会大会プログラム・講演要旨集, 95回, 2P, 260, 2022年11月
(公社)日本生化学会, 日本語

新規クロザピン結合タンパク質の探索 α/βチューブリンに対する相互作用　　　　　　　　　　　　　　　
國井 泰人, 日野 瑞城, 近藤 豪, 和田 明, 松本 純弥, 丹羽 真一, 瀬藤 光利, 矢部 博興, 精神神経学雑誌, 2020特別号, S306, S306, 2020年09月
(公社)日本精神神経学会, 日本語

新規クロザピン結合タンパク質の探索 α/βチューブリンに対する相互作用　　　　　　　　　　　　　　　
國井 泰人, 日野 瑞城, 近藤 豪, 和田 明, 松本 純弥, 丹羽 真一, 瀬藤 光利, 矢部 博興, 精神神経学雑誌, 2020特別号, S306, S306, 2020年09月
(公社)日本精神神経学会, 日本語

イメージング質量分析の最先端
近藤 豪, 瀬藤光利, 生体の科学, 68, 5, 418, 419, 2017年10月, ［招待有り］
日本語, 記事・総説・解説・論説等（商業誌、新聞、ウェブメディア）

統合失調症患者の前頭葉におけるタンパク質プロファイリングと変動する分子ネットワークの同定　　　　　　　　　　　　　　　
平山 未央, 國井 泰人, 松本 純弥, 和田 明, 日野 瑞城, 矢部 博興, 丹羽 真一, 近藤 豪, 瀬藤 光利, 大槻 純男, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 88回・38回, [2LBA087], [2LBA087], 2015年12月
(公社)日本生化学会, 日本語

最先端質量分析 マスプロファイリングからマスイメージングへ タウリンのイメージング質量分析
松下 祥子, 正木 紀隆, 近藤 豪, 武井 史郎, 赤津 裕康, 秋田 天平, 杉山 栄二, 橋詰 良夫, 福田 敦夫, 瀬藤 光利, 臨床化学, 44, Suppl.1, 167, 167, 2015年10月
(一社)日本臨床化学会, 日本語

前頭側頭葉変性症患者脳内におけるタウリンの濃度および分布解析　　　　　　　　　　　　　　　
松下 祥子, 正木 紀隆, 近藤 豪, 武井 史郎, 赤津 裕康, 秋田 天平, 杉山 栄二, 福田 敦夫, 矢尾 育子, 瀬藤 光利, JSBMS Letters, 40, Suppl., 101, 101, 2015年08月, ［査読有り］
(一社)日本医用マススペクトル学会, 日本語

定量プロテオミクスを用いた統合失調症患者脳で発現変動する細胞内ネットワークの同定
平山未央, 平山未央, 國井泰人, 松本純弥, 和田明, 日野瑞城, 矢部博興, 丹羽真一, 近藤豪, 瀬藤光利, 大槻純男, 大槻純男, 生体膜と薬物の相互作用シンポジウム講演要旨集, 37th, 2015年

医用質量分析ガイドブック　　　　　　　　　　　　　　　
近藤 豪, 瀬藤光利, 疾患のイメージング質量分析
診断と治療社, 2013年12月, 9784787820730, 日本語, ［分担執筆］

Direct regulation of lipid microdomains by clinical medicines.　　　　　　　　　　　　　　　
Takeshi Kondo, Mitsutoshi Setou
International Conference on Lipoquality 2017, 2017年09月22日, 英語, 口頭発表（一般）
［国際会議］

医遺伝学　　　　　　　　　　　　　　　
北海道大学
2021年10月 - 現在

医学総論　　　　　　　　　　　　　　　
北海道大学
2019年04月 - 現在

医学研究法I　　　　　　　　　　　　　　　
北海道大学
2019年04月 - 現在

基本医学総論　　　　　　　　　　　　　　　
北海道大学
2019年04月 - 現在

基本医学研究法I　　　　　　　　　　　　　　　
北海道大学
2019年04月 - 現在

生化学I　　　　　　　　　　　　　　　
北海道大学
2019年04月 - 現在

生化学実習　　　　　　　　　　　　　　　
北海道大学
2018年10月 - 現在

顕微鏡学・質量分析学　　　　　　　　　　　　　　　
浜松医科大学

脳実習　　　　　　　　　　　　　　　
浜松医科大学

系統解剖学　　　　　　　　　　　　　　　
浜松医科大学

組織学　　　　　　　　　　　　　　　
浜松医科大学

解剖学総論　　　　　　　　　　　　　　　
浜松医科大学

抗Sez6l2抗体関連脳炎の病態解明
科学研究費助成事業
2023年04月01日 - 2026年03月31日
矢口 裕章, 松島 理明, 近藤 豪, 高橋 秀尚
日本学術振興会, 基盤研究(C), 北海道大学, 23K06940

新しい脂質解析技術を用いた脳脂質異常の原因と意義の解明
科学研究費助成事業
2022年04月01日 - 2025年03月31日
近藤 豪
日本学術振興会, 基盤研究(C), 北海道大学, 22K06875

新規に開発した基質同定法を用いたがんドライバーユビキチンリガーゼの機能解析
科学研究費助成事業
2021年04月01日 - 2024年03月31日
渡部 昌, 近藤 豪
(1)がん関連ユビキチンリガーゼ基質同定プローブの作製と安定発現細胞株の樹立：解析対象の１８種類のユビキチンリガーゼ遺伝子を入手し、基質を捕獲するためのユビキチンリガーゼプローブをレトロウイルス発現ベクター上に組み込んだ。今年度は変異していない野生型のユビキチンリガーゼ遺伝子について作製を行った。作製したプローブベクターを用いてレトロウイルスを作製し、プローブを安定に発現する細胞株作製を試みたところ、１３種類のプローブについて作製に成功した。残り５種については細胞毒性のために作製できなかった。
(2) イオントラップ・オービトラップ型質量分析器による基質・結合分子同定：樹立した１３種類の細胞からユビキチン化タンパク質を精製し、高感度質量分析計にて網羅的な同定を行った。具体的には、それぞれの細胞を可溶化し、抗FLAG抗体で免疫沈降を行い、第一段階の精製を行った。トリクロロ酢酸を用いて沈殿後、アセトンによる洗浄、乾燥、ジチオスレイトールにて還元、ヨードアセタミドにてアルキル化を行った後にトリプシンにてペプチドへと分解後、脱塩処理を行った。さらに抗ユビキチンレムナント抗体で再度免疫沈降を行い、第二段階の精製を行い、再度脱塩処理を行った。得られたサンプルについて質量分析を行った。実験は全て３回繰り返した。得られた結果は、過去に我々が行い蓄積している同様の基質同定結果と比較してスコアを算出することで、個々のユビキチンリガーゼ特異的な基質候補を抽出した。抗ユビキチンレムナント抗体について、これまではアガロースビーズに結合したものを使用していたが、今年度はマグネットビーズに結合した抗体についても検討を行い、アガロースビーズと同等またはそれ以上の結果が得られることを確認した。
日本学術振興会, 基盤研究(B), 北海道大学, 21H02690

光による脂質の同定制御観察技術すなわちオプトリピドミクスの創生　　　　　　　　　　　　　　　
革新的先端研究開発支援事業ユニットタイプ（AMED-CREST）
2015年12月 - 2021年03月
瀬藤光利
国立研究開発法人 日本医療研究開発機構, 競争的資金

精神疾患の分子病態における脂質の役割の解明　　　　　　　　　　　　　　　
科学研究費補助金（若手研究）
2018年 - 2021年
近藤 豪
文部科学省, 研究代表者, 競争的資金

質量顕微鏡法を用いた新しい薬物動態解析及び創薬標的探索事業　　　　　　　　　　　　　　　
研究成果展開事業 大学発新産業創出プログラム
2017年10月 - 2019年03月
瀬藤光利
国立研究開発法人 科学技術振興機構, 競争的資金

細胞骨格制御によるミトコンドリア品質維持機構の解明　　　　　　　　　　　　　　　
科学研究費補助金（若手研究(B)）
2015年 - 2017年
近藤 豪
文部科学省, 研究代表者, 競争的資金

質量顕微鏡法による精神神経疾患脳における脂質‐炎症相互連関の解明　　　　　　　　　　　　　　　
科学研究費補助金（特別研究員奨励費）
2014年 - 2016年
近藤 豪
文部科学省, 研究代表者, 競争的資金

標的物質と親和性の高い物質をスクリーニングする方法　　　　　　　　　　　　　　　
特許権, 瀬藤光利, 近藤豪
特願2017-016609, 2017年02月01日