研究者基本情報

• https://researchmap.jp/daikisetoyama

• https://jglobal.jst.go.jp/detail?JGLOBAL_ID=202601001817587131

• 30550850

• 202601001817587131

研究活動情報

• 2021年, 令和3年日本医療検査科学会優秀演題賞
• 2019年, 令和元年日本医療検査科学会優秀演題賞
• 2014年, 平成26年度がん研究助成金優秀賞

• An Orally Available Derivative of a Specific CK2 Inhibitor for Regulating Circadian Rhythms and Acute Myeloid Leukemia
  Oshima T.; Setoyama D.; co-first author; Sugiyama A.; Amaike K.; Yamanaka I.; Hosokawa K.; Nunomura K.; Kawamura K.; Srivastava A.; Iida M.; Gotoh K.; Lin B.; Morita M.; Ohmi M.; Uchiumi T.; Arai F.; Haruta J.; Sudo M.; Tama F.; Kunisaki Y.; Itami K.; Hirota T.
  Precision Cheistry., 2026年04月, ［査読有り］, ［筆頭著者］
• Fully Automated Serum LC-MS/MS Platform and Pediatric Reference Intervals for Organic Acids, Amino Acids, and Acylcarnitines in Children (Ages 0-6 Years): Toward Quantitative Diagnosis of Inborn Errors of Metabolism.
  Yasushi Ueyanagi; Daiki Setoyama; Tsuyoshi Nakanishi; Yuichi Mushimoto; Vlad Tocan; Hironori Kobayashi; Miki Matsui; Shinya Matsumoto; Akiyoshi Fujishima; Taeko Hotta; Ayumi Sakata; Yuya Kunisaki
  Diagnostics (Basel, Switzerland), 16, 6, 2026年03月19日, ［査読有り］, ［責任著者］, ［国際誌］
  英語, 研究論文（学術雑誌）, Background/Objectives: Conventional diagnosis of inborn errors of metabolism (IEMs) requires multiple specimen types-urine organic acids, plasma amino acids, and serum acylcarnitines-analyzed on distinct analytical platforms. This multi-assay approach is labor-intensive and limits timely clinical decision making. We aimed to develop a fully automated serum-based LC-MS/MS platform for integrated quantitative metabolite profiling and to establish pediatric reference intervals (RIs) to support diagnostic interpretation. Methods: A fully automated LC-MS/MS system integrated with the CLAM-2030 automated pretreatment module was developed to enable simultaneous quantification of 25 organic acids, 8 amino acids, and 21 acylcarnitines. Analytical performance was assessed for linearity, limits of detection and quantification, precision and accuracy. Serum samples from 296 non-IEM children aged 0-6 years were analyzed to establish pediatric RIs using Box-Cox transformation and Gaussian modeling. Clinical utility was evaluated in sera from 89 patients diagnosed with IEM using z-score-based logistic regression models. Results: The method demonstrated excellent performance, with linearity (r2 > 0.99) across calibration ranges, limits of detection and quantification defined by signal-to-noise ratios > 3 and >10, and intra- and inter-assay precision < 15% CV for all 54 analytes. Twenty-one analytes met the acceptance criterion of ±20% accuracy at all quality-control levels. Pediatric RIs provided a quantitative framework for interpreting the metabolic abnormalities. In IEM patients, disease-specific metabolites were consistently outside the established ranges, and z-score-based logistic regression models successfully distinguished major IEM categories, including organic acidemias and long-chain fatty acid oxidation disorders. Conclusions: This fully automated, serum-based LC-MS/MS platform provides a clinically practical and quantitative framework for integrated metabolic profiling using pediatric RIs, supporting diagnosis and monitoring of IEMs in pediatric settings.
• Differences of Plasma Metabolites in Sarcopenia Among Individuals with Type 2 Diabetes: A Cross-Sectional Study.
  Junglyun Kim; Jieun Lee; Daiki Setoyama; Hyon-Seung Yi
  Current medicinal chemistry, 2026年01月13日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, INTRODUCTION: The objective of this study is to characterize plasma metabolites and identify potential clinical indicators of sarcopenia in individuals with Type 2 Diabetes (T2D). METHODS: This cross-sectional, observational study included 122 individuals with T2D recruited from a university hospital clinic between October 2022 and December 2023. Demographic and health data were obtained through questionnaires and electronic health records. Plasma metabolites were analyzed using liquid chromatography-mass spectrometry. Predictive probability of the identified metabolite was assessed using the Receiver Operating Characteristic-Area Under the Curve (ROC-AUC). Statistical analyses were conducted with SPSS version 29 and R version 4.4.2. RESULTS: We identified 23 metabolites, including 21 that were unique to individuals with sarcopenia compared with the general population. Plasma 4-Hydroxyproline and homocysteine levels were significantly different between the sarcopenia and non-sarcopenia groups, as defined by the Asian Working Group for Sarcopenia criteria. A model combining 4-Hydroxyproline, age, body mass index, and educational levels achieved an ROC-AUC of 0.825 (95% CI: 0.726-0.923) for predicting sarcopenia. DISCUSSION: The metabolic features of sarcopenic status are distinctive among individuals with T2D compared to the general population. Plasma 4-Hydroxyproline and Homocysteine are clinical indicators in sarcopenia in T2D. CONCLUSIONS: Individuals with T2D and sarcopenia exhibit distinct metabolite profiles compared with the general population. Plasma 4-Hydroxyproline and homocysteine may serve as clinical indicators for sarcopenia in T2D. Further studies are needed to validate these findings and assess their potential as diagnostic and therapeutic targets.
• Non-canonical role of natural quinones in mitochondrial nucleoid organization for maintaining respiration and protecting cardiac function.
  Soumyadip Pal; Takaya Ishihara; Daiki Setoyama; Chang-Lin Chen; Kenta Onoue; Shigenobu Yonemura; Emi Ogasawara; Naotada Ishihara
  Journal of biochemistry, 179, 1, 5, 20, 2026年01月12日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Mitochondria contain their own DNA (mtDNA), which is essential for respiratory function. Multiple copies of mtDNA are assembled into dot-like structures called nucleoids. Nucleoids move dynamically within mitochondria, and their size and distribution are influenced by mitochondrial membrane fission and fusion. However, the molecular mechanisms and their pathophysiological significance, particularly in vivo, remain largely unknown. Here, we identify a novel role for ubiquinone, as well as natural quinones lacking electron-carrying capacity, in the organization of nucleoids and respiratory complexes, independent of their conventional roles. These quinones facilitate the association and packaging of mtDNA on the cardiolipin-enriched mitochondrial inner membrane. This quinone-dependent maintenance of nucleoids protects against mitochondrial dysfunction and heart failure induced by the anticancer drug doxorubicin. Our RNAi screen identifies a set of genes involved in mitochondrial diseases that exhibit nucleoid deformation, suggesting a novel therapeutic approach targeting mitochondrial nucleoids for various pathological conditions associated with mitochondrial dysfunction.
• Leveraging machine learning to uncover the hidden links between trusting behavior and biological markers.
  Zimu Cao; Daiki Setoyama; Monica Natsumi Daudelin; Toshio Matsushima; Yuichiro Yada; Motoki Watabe; Takatoshi Hikida; Takahiro A Kato; Honda Naoki
  Dialogues in clinical neuroscience, 27, 1, 201, 215, 2025年12月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Understanding the decision-making mechanisms underlying trust is essential, particularly for individuals with mental disorders who often experience difficulties in forming interpersonal trust. In this study, we aimed to explore biomarkers associated with trust-based decision-making through quantitative analysis. However, quantifying internal decision-making processes is challenging, as they are not directly observable. To address this, we developed a machine learning method based on a Bayesian hierarchical model to quantitatively infer latent decision-making parameters from behavioural data collected during a trust game. Applying this method to data from patients with major depressive disorder (MDD) and healthy controls (HCs), we estimated individualised model parameters that regulate trust-related decisions. The model successfully predicted participants' behaviours in the task. Although no significant group-level differences were observed in the estimated parameters between the MDD and HC groups, we uncovered hidden links between trust-related decision-making processes and specific blood biomarkers. Notably, metabolites such as 5-aminolevulinic acid, acetylcarnitine, and 2-aminobutyric acid were significantly associated with individual differences in trusting behaviour. These findings provide valuable insight into the biological basis of trust-based decision-making. They also offer a novel framework for integrating behavioural modelling with biomarker discovery, potentially informing the development of targeted interventions to enhance social functioning and overall well-being.
• Acute and long-term effects of repeated ketamine infusions in treatment-resistant depression and associated metabolite changes.
  Hitoshi Sakurai; Daiki Setoyama; Takahiro A Kato; Hisateru Tachimori; Masami Murao; Yasuyuki Matsumoto; Teruo Tada; Yayoi Imamura; Hiroyuki Seki; Takashi Tsuboi; Hiroyuki Uchida; Koichiro Watanabe
  Psychiatry and clinical neurosciences, 79, 10, 645, 652, 2025年10月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, AIM: This study aims to investigate the acute and sustained antidepressant effects of repeated ketamine infusions in patients with treatment-resistant depression (TRD), and to identify early metabolomic changes predictive of treatment outcomes using metabolome analyses. METHODS: This open-label study investigated the effects of four intravenous ketamine infusions (0.5 mg/kg) administered over 2 weeks in 30 patients with TRD. Clinical outcomes, including the Montgomery-Åsberg Depression Rating Scale (MADRS), were evaluated at baseline, 1-2 h after each infusion, and monthly during a 12-month follow-up. Plasma levels of 11 depression-associated metabolites were measured at baseline and 1.5-2 h after the first infusion. A general linear model was employed to analyze the association between metabolite changes after the first infusion and the MADRS score percent improvements after the fourth infusion and at 12 months. Remission was defined as a MADRS score of ≤10. RESULTS: All participants completed the four infusions. The MADRS score decreased from 30.6 ± 6.1 at baseline to 20.3 ± 11.5 after the fourth infusion, with a remission rate of 26.7%. At 12 months, 13.3% of participants remained in remission. Changes in 3-hydroxybutyrate levels after the first infusion predicted the MADRS score percent improvements after the fourth infusion (β = 1.35, 95% CI: 0.41-2.30, P = 0.005) and at 12 months (β = 1.38, 95% CI: 0.37-2.39, P = 0.007). CONCLUSION: While repeated ketamine infusions demonstrated rapid antidepressant effects, sustained remission was achieved in a minority of patients with TRD. 3-Hydroxybutyrate may serve as a biomarker for predicting treatment response. These findings underscore the potential for individualized strategies using ketamine infusions. CLINICAL TRIAL REGISTRATION: jRCTs031210648 (Japan Registry of Clinical Trials).
• Glutamate in cerebrospinal fluid as a diagnostic marker for acute encephalopathy in childhood.
  Kenta Kajiwara; Daiki Setoyama; Kanako Higashi; Tomoko Nomiyama; Yuko Ichimiya; Daichi Kumamoto; Satoshi Akamine; Yuri Sonoda; Pin Fee Chong; Ryuichi Takemoto; Wakato Matsuoka; Soichi Mizuguchi; Noriyuki Kaku; Takahiro A Kato; Tomohiko Akahoshi; Yuya Kunisaki; Yasunari Sakai; Shouichi Ohga
  Brain & development, 47, 5, 104448, 104448, 2025年10月, ［査読有り］, ［筆頭著者］, ［国際誌］
  英語, 研究論文（学術雑誌）, BACKGROUNDS: Acute encephalopathy with biphasic seizures and late reduced diffusion (AESD) is the most frequent form of acute encephalopathy in early childhood in Japan. Magnetic resonance imaging provides useful hallmarks of diagnosing AESD. However, metabolomic profiles for AESD remain elusive. This study investigates whether measurement of amino acids in the cerebrospinal fluid (CSF) is useful for the diagnosis of AESD before onset. METHODS: In the first study, CSF samples were collected from patients (11 AESD and 17 controls) admitted to Kyushu University Hospital during 2011-2016. Amino acids in the CSF were analyzed using mass spectrometry. Cytometric bead arrays were used to measure cytokine and chemokine levels in the CSF. The second study was performed by recruiting patients (8 AESD patients and 10 controls) admitted during 2011-2024. CSF samples were stored at -20 °C for 1 month to 12 years. RESULTS: In the first study, glutamate levels in the CSF from AESD patients were higher than in controls and correlated with methionine, threonine, and tyrosine levels. A correlation map of cytokines and amino acids revealed that glutamate formed a cluster with IL-1β, IL-10, and IL-12 p70. In the second study, no difference in glutamate levels was observed between AESD and control groups. CONCLUSIONS: CSF glutamate potentially serves as a useful marker for diagnosing AESD. The long-term storage of CSF samples was likely to cause a decay of glutamate in the CSF. Prospective studies using fresh CSF samples are necessary to validate the results in this study.
• AVID mouse: A versatile platform for real-time, multiscale ATP imaging and spatial systems metabolism analysis in living mice.
  Yuichiro Ohnishi; Daiki Setoyama; Hideki Miwa; Norimichi Koitabashi; Riki Ogasawara; Seri Kitada; Naoki Matoba; Takahito Ayano; Kaoru Hiramoto; Ryuto Yasui; Yuki Sugiura; Takahisa Anada; Kosuke Ino; Hinako Matsuda; Takahiro Noma; Shigenori Nonaka; Takashi Izumi; Masahiko Kurabayashi; Makoto Suematsu; Yuya Kunisaki; Motoko Yanagita; Hiromi Imamura; Masamichi Yamamoto
  Cell reports, 44, 9, 116246, 116246, 2025年09月23日, ［査読有り］, ［筆頭著者］, ［国際誌］
  英語, 研究論文（学術雑誌）, We developed the AVID (ATP visualization in vivo directly) mouse, a genetically encoded biosensor mouse enabling real-time, multiscale imaging of ATP dynamics across the whole body, organs, and cellular compartments in living animals. AVID revealed previously undetectable localized ATP depletion near the central vein of the liver after myocardial infarction, spatially linked to kynurenic acid accumulation-a phenomenon invisible to conventional bulk metabolomics. By seamlessly integrating macroscopic organ-level imaging with microscopic spatial metabolomics, AVID establishes a new framework for spatial systems metabolism. Beyond myocardial infarction, this platform offers broad applicability to study organ-organ metabolic communication, spatial metabolic heterogeneity, and localized metabolic shifts across diverse physiological and pathological contexts, providing a transformative resource for metabolic research.
• A Nucleic Acid Prodrug That Activates Mitochondrial Respiration, Promotes Stress Resilience, and Prolongs Lifespan.
  Takahisa Anada; Michiharu Kawahara; Taisei Shimada; Ryotaro Kuroda; Hidenori Okamura; Daiki Setoyama; Fumi Nagatsugi; Yuya Kunisaki; Eriko Kage-Nakadai; Shingo Kobayashi; Masaru Tanaka
  Journal of the American Chemical Society, 147, 25, 22161, 22175, 2025年06月25日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Mitochondrial dysfunction caused by aging leads to decreased energy metabolism, resulting in functional decline and increased frailty in multiple tissues. Strategies for protecting and activating mitochondria under stressful conditions are required to suppress aging and age-related diseases. However, it is challenging to develop drugs capable of boosting mitochondrial respiration and compensating for the reduced intracellular adenosine triphosphate (ATP) levels. In this study, we developed a prodrug that stimulates the metabolism of intracellular adenine nucleotides (AXP: adenosine monophosphate (AMP), adenosine diphosphate (ADP), and ATP). It enhances AMP-activated protein kinase activity, fatty acid oxidation, oxidative stress resistance, and mitochondrial respiration, thereby increasing the intracellular ATP levels. Furthermore, this prodrug markedly extended the lifespan of Caenorhabditis elegans. AXP-driven stimulation of cellular energy metabolism proposed herein represents a novel geroprotective strategy and paves the way for the development of bioenergetic-molecule therapeutics.
• Cerebrospinal fluid proteomics exerts predictive potential for immune effector cell-associated neurotoxicity syndrome (ICANS) in CAR-T cell therapy.
  Tomoko Nomiyama; Daiki Setoyama; Ikumi Yamanaka; Masatoshi Shimo; Kohta Miyawaki; Takuji Yamauchi; Fumiaki Jinnouchi; Teppei Sakoda; Kensuke Sasaki; Takahiro Shima; Yoshikane Kikushige; Yasuo Mori; Koichi Akashi; Koji Kato; Yuya Kunisaki
  Leukemia, 39, 4, 983, 987, 2025年04月, ［査読有り］, ［筆頭著者］, ［国際誌］
  英語, 研究論文（学術雑誌）
• Comparative Analysis of Primary Sarcopenia and End-Stage Renal Disease-Related Muscle Wasting Using Multi-Omics Approaches.
  Daiki Setoyama; Dohyun Han; Jingwen Tian; Ho Yeop Lee; Hyun Suk Shin; Ha Thi Nga; Thi Linh Nguyen; Ji Sun Moon; Hyo Ju Jang; Evonne Kim; Seong-Kyu Choe; Sang Hyeon Ju; Dae Eun Choi; Obin Kwon; Hyon-Seung Yi
  Journal of cachexia, sarcopenia and muscle, 16, 2, e13749, 2025年04月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, BACKGROUND: Age-related primary sarcopenia and end-stage renal disease (ESRD)-related muscle wasting are discrete entities; however, both manifest as a decline in skeletal muscle mass and strength. The etiological pathways differ, with aging factors implicated in sarcopenia and a combination of uremic factors, including haemodialysis, contributing to ESRD-related muscle wasting. Understanding these molecular nuances is imperative for targeted interventions, and the integration of proteomic and metabolomic data elucidate these intricate processes. METHODS: We generated detailed clinical data and multi-omics data (plasma proteomics and metabolomics) for 78 participants to characterise sarcopenia (n = 28; mean age, 72.6 ± 7.0 years) or ESRD (n = 22; 61.6 ± 5.5 years) compared with controls (n = 28; 69.3 ± 5.7 years). Muscle mass was measured using bioelectrical impedance analysis and handgrip strength. Five-times sit-to-stand test performance was measured for all participants. Sarcopenia was diagnosed in accordance with the 2019 Consensus Guidelines from the Asian Working Group for Sarcopenia. An abundance of 234 metabolites and 722 protein groups was quantified in all plasma samples using liquid chromatography with tandem mass spectrometry. RESULTS: Muscle mass, handgrip strength and lower limb muscle function significantly lower in the sarcopenia group and the ESRD group compared with those in the control group. Metabolomics revealed altered metabolites, highlighting exclusive differences in ESRD-related muscle wasting. Metabolite set enrichment analysis revealed the involvement of numerous metabolic intermediates associated with urea cycle, amino acid metabolism and nucleic acid metabolism. Catecholamines, including epinephrine, dopamine and serotonin, are significantly elevated in the plasma of patients within the ESRD group. Proteomics data exhibited a clearer distinction among the three groups compared with the metabolomics data, particularly in distinguishing the control group from the sarcopenia group. The ciliary neurotrophic factor receptor was top-ranked in terms of the variable importance of projection scores. Plasma AHNAK protein levels was higher in the sarcopenia group but was lower in the ESRD group. Proteomic set enrichment analysis revealed enrichment of several pathways related to sarcopenia, such as hemopexin, defence response and cell differentiation, in sarcopenia group. Multi-omic integration analysis revealed associations between relevant metabolites, including catecholamines, and a group of annotated proteins in extracellular exosomes. CONCLUSIONS: We identified distinct multi-omic signatures in individuals with ESRD or sarcopenia, providing new insights into the mechanisms underlying ESRD-related muscle wasting, which differ from primary sarcopenia. These findings may support interventions for context-dependent muscle loss and contribute to the development of targeted treatments and preventive strategies for muscle wasting.
• Glutaminolysis is associated with mitochondrial pathway activation and can be therapeutically targeted in glioblastoma.
  Kenji Miki; Mikako Yagi; Ryusuke Hatae; Ryosuke Otsuji; Takahiro Miyazaki; Katsuhiro Goto; Daiki Setoyama; Yutaka Fujioka; Yuhei Sangatsuda; Daisuke Kuga; Nayuta Higa; Tomoko Takajo; Yonezawa Hajime; Toshiaki Akahane; Akihide Tanimoto; Ryosuke Hanaya; Yuya Kunisaki; Takeshi Uchiumi; Koji Yoshimoto
  Cancer & metabolism, 12, 1, 35, 35, 2024年11月19日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, BACKGROUND: Glioblastoma is an aggressive cancer that originates from abnormal cell growth in the brain and requires metabolic reprogramming to support tumor growth. Metabolic reprogramming involves the upregulation of various metabolic pathways. Although the activation of specific metabolic pathways in glioblastoma cell lines has been documented, the comprehensive profile of metabolic reprogramming and the role of each pathway in glioblastoma tissues in patients remain elusive. METHODS: We analyzed 38 glioblastoma tissues. As a test set, we examined 20 tissues from Kyushu University Hospital, focusing on proteins related to several metabolic pathways, including glycolysis, the one-carbon cycle, glutaminolysis, and the mitochondrial tricarboxylic acid cycle. Subsequently, we analyzed an additional 18 glioblastoma tissues from Kagoshima University Hospital as a validation set. We also validated our findings using six cell lines, including U87, LN229, U373, T98G, and two patient-derived cells. RESULTS: The levels of mitochondria-related proteins (COX1, COX2, and DRP1) were correlated with each other and with glutaminolysis-related proteins (GLDH and GLS1). Conversely, their expression was inversely correlated with that of glycolytic proteins. Notably, inhibiting the glutaminolysis pathway in cell lines with high GLDH and GLS1 expression proved effective in suppressing tumor growth. CONCLUSIONS: Our findings confirm that glioblastoma tissues can be categorized into glycolytic-dominant and mitochondrial-dominant types, as previously reported. The mitochondrial-dominant type is also glutaminolysis-dominant. Therefore, inhibiting the glutaminolysis pathway may be an effective treatment for mitochondrial-dominant glioblastoma.
• An N-terminal and ankyrin repeat domain interactome of Shank3 identifies the protein complex with the splicing regulator Nono in mice.
  Sayaka Okuzono; Fumihiko Fujii; Daiki Setoyama; Ryoji Taira; Yohei Shinmyo; Hiroki Kato; Keiji Masuda; Kousuke Yonemoto; Satoshi Akamine; Yuki Matsushita; Yoshitomo Motomura; Takeshi Sakurai; Hiroshi Kawasaki; Kihoon Han; Takahiro A Kato; Hiroyuki Torisu; Dongchon Kang; Yusaku Nakabeppu; Shouichi Ohga; Yasunari Sakai
  Genes to cells : devoted to molecular & cellular mechanisms, 29, 9, 746, 756, 2024年09月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, An autism-associated gene Shank3 encodes multiple splicing isoforms, Shank3a-f. We have recently reported that Shank3a/b-knockout mice were more susceptible to kainic acid-induced seizures than wild-type mice at 4 weeks of age. Little is known, however, about how the N-terminal and ankyrin repeat domains (NT-Ank) of Shank3a/b regulate multiple molecular signals in the developing brain. To explore the functional roles of Shank3a/b, we performed a mass spectrometry-based proteomic search for proteins interacting with GFP-tagged NT-Ank. In this study, NT-Ank was predicted to form a variety of complexes with a total of 348 proteins, in which RNA-binding (n = 102), spliceosome (n = 22), and ribosome-associated molecules (n = 9) were significantly enriched. Among them, an X-linked intellectual disability-associated protein, Nono, was identified as a NT-Ank-binding protein. Coimmunoprecipitation assays validated the interaction of Shank3 with Nono in the mouse brain. In agreement with these data, the thalamus of Shank3a/b-knockout mice aberrantly expressed splicing isoforms of autism-associated genes, Nrxn1 and Eif4G1, before and after seizures with kainic acid treatment. These data indicate that Shank3 interacts with multiple RNA-binding proteins in the postnatal brain, thereby regulating the homeostatic expression of splicing isoforms for autism-associated genes after birth.
• Brown fat-specific mitoribosomal function is crucial for preventing cold exposure-induced bone loss.
  Jingwen Tian; Ji Sun Moon; Ha Thi Nga; Ho Yeop Lee; Thi Linh Nguyen; Hyo Ju Jang; Daiki Setoyama; Minho Shong; Ju Hee Lee; Hyon-Seung Yi
  Cellular and molecular life sciences : CMLS, 81, 1, 314, 314, 2024年07月27日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, This study examines the interplay between ambient temperature, brown adipose tissue (BAT) function, and bone metabolism, emphasizing the effects of cold exposure and BAT mitochondrial activity on bone health. Utilizing ovariectomized (OVX) mice to model primary osteoporosis and BAT-specific mitochondrial dysfunction (BKO) mice, we evaluated the impact of housing temperature on bone density, immune modulation in bone marrow, and the protective role of BAT against bone loss. Cold exposure was found to universally reduce bone mass, enhance osteoclastogenesis, and alter bone marrow T-cell populations, implicating the immune system in bone remodeling under cold stress. The thermogenic function of BAT, driven by mitochondrial oxidative phosphorylation, was crucial in protecting against bone loss. Impaired BAT function, through surgical removal or mitochondrial dysfunction, exacerbated bone loss in cold environments, highlighting BAT's metabolic role in maintaining bone health. Furthermore, cold-induced changes in BAT function led to systemic metabolic shifts, including elevated long-chain fatty acids, which influenced osteoclast differentiation and activity. These findings suggest a systemic mechanism connecting environmental temperature and BAT metabolism with bone physiology, providing new insights into the metabolic and environmental determinants of bone health. Future research could lead to novel bone disease therapies targeting these pathways.
• ATP1A3 regulates protein synthesis for mitochondrial stability under heat stress.
  Fumihiko Fujii; Hikaru Kanemasa; Sayaka Okuzono; Daiki Setoyama; Ryoji Taira; Kousuke Yonemoto; Yoshitomo Motomura; Hiroki Kato; Keiji Masuda; Takahiro A Kato; Shouichi Ohga; Yasunari Sakai
  Disease models & mechanisms, 17, 6, 2024年06月01日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Pathogenic variants in ATP1A3, the gene encoding the α3 subunit of the Na+/K+-ATPase, cause alternating hemiplegia of childhood (AHC) and related disorders. Impairments in Na+/K+-ATPase activity are associated with the clinical phenotype. However, it remains unclear whether additional mechanisms are involved in the exaggerated symptoms under stressed conditions in patients with AHC. We herein report that the intracellular loop (ICL) of ATP1A3 interacted with RNA-binding proteins, such as Eif4g (encoded by Eif4g1), Pabpc1 and Fmrp (encoded by Fmr1), in mouse Neuro2a cells. Both the siRNA-mediated depletion of Atp1a3 and ectopic expression of the p.R756C variant of human ATP1A3-ICL in Neuro2a cells resulted in excessive phosphorylation of ribosomal protein S6 (encoded by Rps6) and increased susceptibility to heat stress. In agreement with these findings, induced pluripotent stem cells (iPSCs) from a patient with the p.R756C variant were more vulnerable to heat stress than control iPSCs. Neurons established from the patient-derived iPSCs showed lower calcium influxes in responses to stimulation with ATP than those in control iPSCs. These data indicate that inefficient protein synthesis contributes to the progressive and deteriorating phenotypes in patients with the p.R756C variant among a variety of ATP1A3-related disorders.
• Cardiomyocyte-specific deletion of the mitochondrial transporter Abcb10 causes cardiac dysfunction via lysosomal-mediated ferroptosis.
  Yura Do; Mikako Yagi; Haruka Hirai; Kenji Miki; Yukina Fukahori; Daiki Setoyama; Masatatsu Yamamoto; Tatsuhiko Furukawa; Yuya Kunisaki; Dongchon Kang; Takeshi Uchiumi
  Bioscience reports, 44, 5, 2024年05月29日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Heart function is highly dependent on mitochondria, which not only produce energy but also regulate many cellular functions. Therefore, mitochondria are important therapeutic targets in heart failure. Abcb10 is a member of the ABC transporter superfamily located in the inner mitochondrial membrane and plays an important role in haemoglobin synthesis, biliverdin transport, antioxidant stress, and stabilization of the iron transporter mitoferrin-1. However, the mechanisms underlying the impairment of mitochondrial transporters in the heart remain poorly understood. Here, we generated mice with cardiomyocyte-specific loss of Abcb10. The Abcb10 knockouts exhibited progressive worsening of cardiac fibrosis, increased cardiovascular risk markers and mitochondrial structural abnormalities, suggesting that the pathology of heart failure is related to mitochondrial dysfunction. As the mitochondrial dysfunction was observed early but mildly, other factors were considered. We then observed increased Hif1α expression, decreased NAD synthase expression, and reduced NAD+ levels, leading to lysosomal dysfunction. Analysis of ABCB10 knockdown HeLa cells revealed accumulation of Fe2+ and lipid peroxides in lysosomes, leading to ferroptosis. Lipid peroxidation was suppressed by treatment with iron chelators, suggesting that lysosomal iron accumulation is involved in ferroptosis. We also observed that Abcb10 knockout cardiomyocytes exhibited increased ROS production, iron accumulation, and lysosomal hypertrophy. Our findings suggest that Abcb10 is required for the maintenance of cardiac function and reveal a novel pathophysiology of chronic heart failure related to lysosomal function and ferroptosis.
• Collagen Lattice Model, Populated with Heterogeneous Cancer-Associated Fibroblasts, Facilitates Advanced Reconstruction of Pancreatic Cancer Microenvironment.
  Xiaoyu Song; Yuma Nihashi; Yukiko Imai; Nobuhito Mori; Noritaka Kagaya; Hikaru Suenaga; Kazuo Shin-Ya; Masamichi Yamamoto; Daiki Setoyama; Yuya Kunisaki; Yasuyuki S Kida
  International journal of molecular sciences, 25, 7, 2024年03月27日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Pancreatic ductal adenocarcinoma (PDAC) is a solid-tumor malignancy. To enhance the treatment landscape of PDAC, a 3D model optimized for rigorous drug screening is essential. Within the PDAC tumor microenvironment, a dense stroma comprising a large extracellular matrix and cancer-associated fibroblasts (CAFs) is well-known for its vital role in modulating tumor growth, cellular heterogeneity, bidirectional paracrine signaling, and chemoresistance. In this study, we employed a fibroblast-populated collagen lattice (FPCL) modeling approach that has the ability to replicate fibroblast contractility in the collagenous matrix to build dense stroma. This FPCL model allows CAF differentiation by facilitating multifaceted cell-cell interactions between cancer cells and CAFs, with the differentiation further influenced by mechanical forces and hypoxia carried within the 3D structure. Our FPCL models displayed hallmark features, including ductal gland structures and differentiated CAFs with spindle shapes. Through morphological explorations alongside in-depth transcriptomic and metabolomic profiling, we identified substantial molecular shifts from the nascent to mature model stages and potential metabolic biomarkers, such as proline. The initial pharmacological assays highlighted the effectiveness of our FPCL model in screening for improved therapeutic strategies. In conclusion, our PDAC modeling platform mirrors complex tumor microenvironmental dynamics and offers an unparalleled perspective for therapeutic exploration.
• Exploring the Role of Desmoplastic Physical Stroma in Pancreatic Cancer Progression Using a Three-Dimensional Collagen Matrix Model.
  Xiaoyu Song; Yuma Nihashi; Masamichi Yamamoto; Daiki Setoyama; Yuya Kunisaki; Yasuyuki S Kida
  Bioengineering (Basel, Switzerland), 10, 12, 2023年12月18日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Pancreatic ductal adenocarcinoma (PDAC) is a refractory tumor with a poor prognosis, and its complex microenvironment is characterized by a fibrous interstitial matrix surrounding PDAC cells. Type I collagen is a major component of this interstitial matrix. Abundant type I collagen promotes its deposition and cross-linking to form a rigid and dense physical barrier, which limits drug penetration and immune cell infiltration and provides drug resistance and metabolic adaptations. In this study, to identify the physical effect of the stroma, type I collagen was used as a 3D matrix to culture Capan-1 cells and generate a 3D PDAC model. Using transcriptome analysis, a link between type I collagen-induced physical effects and the promotion of Capan-1 cell proliferation and migration was determined. Moreover, metabolomic analysis revealed that the physical effect caused a shift in metabolism toward a glycolytic phenotype. In particular, the high expression of proline in the metabolites suggests the ability to maintain Capan-1 cell proliferation under hypoxic and nutrient-depleted conditions. In conclusion, we identified type I collagen-induced physical effects in promoting Capan-1 cells, which cause PDAC progression, providing support for the role of dense stroma in the PDAC microenvironment and identifying a fundamental method for modeling the complex PDAC microenvironment.
• Improving lysosomal ferroptosis with NMN administration protects against heart failure.
  Mikako Yagi; Yura Do; Haruka Hirai; Kenji Miki; Takahiro Toshima; Yukina Fukahori; Daiki Setoyama; Chiaki Abe; Yo-Ichi Nabeshima; Dongchon Kang; Takeshi Uchiumi
  Life science alliance, 6, 12, 2023年12月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Myocardial mitochondria are primary sites of myocardial energy metabolism. Mitochondrial disorders are associated with various cardiac diseases. We previously showed that mice with cardiomyocyte-specific knockout of the mitochondrial translation factor p32 developed heart failure from dilated cardiomyopathy. Mitochondrial translation defects cause not only mitochondrial dysfunction but also decreased nicotinamide adenine dinucleotide (NAD+) levels, leading to impaired lysosomal acidification and autophagy. In this study, we investigated whether nicotinamide mononucleotide (NMN) administration, which compensates for decreased NAD+ levels, improves heart failure because of mitochondrial dysfunction. NMN administration reduced damaged lysosomes and improved autophagy, thereby reducing heart failure and extending the lifespan in p32cKO mice. We found that lysosomal damage due to mitochondrial dysfunction induced ferroptosis, involving the accumulation of iron in lysosomes and lipid peroxide. The ameliorative effects of NMN supplementation were found to strongly affect lysosomal function rather than mitochondrial function, particularly lysosome-mediated ferroptosis. NMN supplementation can improve lysosomal, rather than mitochondrial, function and prevent chronic heart failure.
• Mitochondrial haplotype mutation alleviates respiratory defect of MELAS by restoring taurine modification in tRNA with 3243A > G mutation.
  Saori Ueda; Mikako Yagi; Ena Tomoda; Shinya Matsumoto; Yasushi Ueyanagi; Yura Do; Daiki Setoyama; Yuichi Matsushima; Asuteka Nagao; Tsutomu Suzuki; Tomomi Ide; Yusuke Mori; Noriko Oyama; Dongchon Kang; Takeshi Uchiumi
  Nucleic acids research, 51, 14, 7480, 7495, 2023年08月11日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, The 3243A > G in mtDNA is a representative mutation in mitochondrial diseases. Mitochondrial protein synthesis is impaired due to decoding disorder caused by severe reduction of 5-taurinomethyluridine (τm5U) modification of the mutant mt-tRNALeu(UUR) bearing 3243A > G mutation. The 3243A > G heteroplasmy in peripheral blood reportedly decreases exponentially with age. Here, we found three cases with mild respiratory symptoms despite bearing high rate of 3243A > G mutation (>90%) in blood mtDNA. These patients had the 3290T > C haplotypic mutation in addition to 3243A > G pathogenic mutation in mt-tRNALeu(UUR) gene. We generated cybrid cells of these cases to examine the effects of the 3290T > C mutation on mitochondrial function and found that 3290T > C mutation improved mitochondrial translation, formation of respiratory chain complex, and oxygen consumption rate of pathogenic cells associated with 3243A > G mutation. We measured τm5U frequency of mt-tRNALeu(UUR) with 3243A > G mutation in the cybrids by a primer extension method assisted with chemical derivatization of τm5U, showing that hypomodification of τm5U was significantly restored by the 3290T > C haplotypic mutation. We concluded that the 3290T > C is a haplotypic mutation that suppresses respiratory deficiency of mitochondrial disease by restoring hypomodified τm5U in mt-tRNALeu(UUR) with 3243A > G mutation, implying a potential therapeutic measure for mitochondrial disease associated with pathogenic mutations in mt-tRNAs.
• Shank3a/b isoforms regulate the susceptibility to seizures and thalamocortical development in the early postnatal period of mice.
  Sayaka Okuzono; Fumihiko Fujii; Yuki Matsushita; Daiki Setoyama; Yohei Shinmyo; Ryoji Taira; Kousuke Yonemoto; Satoshi Akamine; Yoshitomo Motomura; Masafumi Sanefuji; Takeshi Sakurai; Hiroshi Kawasaki; Kihoon Han; Takahiro A Kato; Hiroyuki Torisu; Dongchon Kang; Yusaku Nakabeppu; Yasunari Sakai; Shouichi Ohga
  Neuroscience research, 193, 13, 19, 2023年08月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Epileptic seizures are distinct but frequent comorbidities in children with autism spectrum disorder (ASD). The hyperexcitability of cortical and subcortical neurons appears to be involved in both phenotypes. However, little information is available concerning which genes are involved and how they regulate the excitability of the thalamocortical network. In this study, we investigate whether an ASD-associated gene, SH3 and multiple ankyrin repeat domains 3 (Shank3), plays a unique role in the postnatal development of thalamocortical neurons. We herein report that Shank3a/b, the splicing isoforms of mouse Shank3, were uniquely expressed in the thalamic nuclei, peaking from two to four weeks after birth. Shank3a/b-knockout mice showed lower parvalbumin signals in the thalamic nuclei. Consistently, Shank3a/b-knockout mice were more susceptible to generalized seizures than wild-type mice after kainic acid treatments. Together, these data indicate that NT-Ank domain of Shank3a/b regulates molecular pathways that protect thalamocortical neurons from hyperexcitability during the early postnatal period of mice.
• IFN-γ-STAT1-ERK Pathway Mediates Protective Effects of Invariant Natural Killer T Cells Against Doxorubicin-Induced Cardiomyocyte Death.
  Masashi Sada; Shouji Matsushima; Masataka Ikeda; Soichiro Ikeda; Kosuke Okabe; Akihito Ishikita; Tomonori Tadokoro; Nobuyuki Enzan; Taishi Yamamoto; Hiroko Deguchi Miyamoto; Yoshitomo Tsutsui; Ryo Miyake; Daiki Setoyama; Dongchon Kang; Tomomi Ide; Hiroyuki Tsutsui
  JACC. Basic to translational science, 8, 8, 992, 1007, 2023年08月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Doxorubicin (DOX)-induced cardiomyopathy has poor prognosis, and myocardial inflammation is intimately involved in its pathophysiology. The role of invariant natural killer T (iNKT) cells has not been fully determined in this disease. We here demonstrated that activation of iNKT cells by α-galactosylceramide (GC) attenuated DOX-induced cardiomyocyte death and cardiac dysfunction. αGC increased interferon (IFN)-γ and phosphorylation of signal transducers and activators of transcription 1 (STAT1) and extracellular signal-regulated kinase (ERK). Administration of anti-IFN-γ neutralizing antibody abrogated the beneficial effects of αGC on DOX-induced cardiac dysfunction. These findings emphasize the protective role of iNKT cells in DOX-induced cardiomyopathy via the IFN-γ-STAT1-ERK pathway.
• Decoding Metabolic Symbiosis between Pancreatic Cancer Cells and Cancer-Associated Fibroblasts Using Cultured Tumor Microenvironment.
  Yuma Nihashi; Xiaoyu Song; Masamichi Yamamoto; Daiki Setoyama; Yasuyuki S Kida
  International journal of molecular sciences, 24, 13, 2023年07月03日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive cancer with a poor prognosis, largely due to its unique tumor microenvironment (TME) and dense fibrotic stroma. Cancer-associated fibroblasts (CAFs) play a crucial role in promoting tumor growth and metastasis, contributing to the metabolic adaptation of PDAC cells. However, the metabolic interactions between PDAC cells and CAFs are not well-understood. In this study, an in vitro co-culture model was used to investigate these metabolic interactions. Metabolomic analysis was performed under monoculture conditions of Capan-1 PDAC cells and CAF precursor cells, as well as co-culture conditions of PDAC cells and differentiated inflammatory CAF (iCAF). Co-cultured Capan-1 cells displayed significant metabolic changes, such as increased 2-oxoglutaric acid and lauric acid and decreased amino acids. The metabolic profiles of co-cultured Capan-1 and CAFs revealed differences in intracellular metabolites. Analysis of extracellular metabolites in the culture supernatant showed distinct differences between Capan-1 and CAF precursors, with the co-culture supernatant exhibiting the most significant changes. A comparison of the culture supernatants of Capan-1 and CAF precursors revealed different metabolic processes while co-culturing the two cell types demonstrated potential metabolic interactions. In conclusion, this study emphasizes the importance of metabolic interactions between cancer cells and CAFs in tumor progression and highlights the role of TME in metabolic reprogramming.
• The SGLT2 inhibitor empagliflozin improves cardiac energy status via mitochondrial ATP production in diabetic mice.
  Jungmi Choi; Naoki Matoba; Daiki Setoyama; Daiki Watanabe; Yuichiro Ohnishi; Ryuto Yasui; Yuichirou Kitai; Aki Oomachi; Yutaro Kotobuki; Yoichi Nishiya; Michael Paul Pieper; Hiromi Imamura; Motoko Yanagita; Masamichi Yamamoto
  Communications biology, 6, 1, 278, 278, 2023年03月17日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Empagliflozin, a sodium-glucose co-transporter 2 inhibitor developed, has been shown to reduce cardiovascular events in patients with type 2 diabetes and established cardiovascular disease. Several studies have suggested that empagliflozin improves the cardiac energy state which is a partial cause of its potency. However, the detailed mechanism remains unclear. To address this issue, we used a mouse model that enabled direct measurement of cytosolic and mitochondrial ATP levels. Empagliflozin treatment significantly increased cytosolic and mitochondrial ATP levels in the hearts of db/db mice. Empagliflozin also enhanced cardiac robustness by maintaining intracellular ATP levels and the recovery capacity in the infarcted area during ischemic-reperfusion. Our findings suggest that empagliflozin enters cardiac mitochondria and directly causes these effects by increasing mitochondrial ATP via inhibition of NHE1 and Nav1.5 or their common downstream sites. These cardioprotective effects may be involved in the beneficial effects on heart failure seen in clinical trials.
• Glycolytic System in Axons Supplement Decreased ATP Levels after Axotomy of the Peripheral Nerve.
  Tomofumi Takenaka; Yuichiro Ohnishi; Masamichi Yamamoto; Daiki Setoyama; Haruhiko Kishima
  eNeuro, 10, 3, 2023年03月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Wallerian degeneration (WD) occurs in the early stages of numerous neurologic disorders, and clarifying WD pathology is crucial for the advancement of neurologic therapies. ATP is acknowledged as one of the key pathologic substances in WD. The ATP-related pathologic pathways that regulate WD have been defined. The elevation of ATP levels in axon contributes to delay WD and protects axons. However, ATP is necessary for the active processes to proceed WD, given that WD is stringently managed by auto-destruction programs. But little is known about the bioenergetics during WD. In this study, we made sciatic nerve transection models for GO-ATeam2 knock-in rats and mice. We presented the spatiotemporal ATP distribution in the injured axons with in vivo ATP imaging systems, and investigated the metabolic source of ATP in the distal nerve stump. A gradual decrease in ATP levels was observed before the progression of WD. In addition, the glycolytic system and monocarboxylate transporters (MCTs) were activated in Schwann cells following axotomy. Interestingly, in axons, we found the activation of glycolytic system and the inactivation of the tricarboxylic acid (TCA) cycle. Glycolytic inhibitors, 2-deoxyglucose (2-DG) and MCT inhibitors, a-cyano-4-hydroxycinnamic acid (4-CIN) decreased ATP and enhanced WD progression, whereas mitochondrial pyruvate carrier (MPC) inhibitors (MSDC-0160) did not change. Finally, ethyl pyruvate (EP) increased ATP levels and delayed WD. Together, our findings suggest that glycolytic system, both in Schwann cells and axons, is the main source of maintaining ATP levels in the distal nerve stump.
• Metabolite Changes during the Transition from Hyperthyroidism to Euthyroidism in Patients with Graves' Disease.
  Ho Yeop Lee; Byeong Chang Sim; Ha Thi Nga; Ji Sun Moon; Jingwen Tian; Nguyen Thi Linh; Sang Hyeon Ju; Dong Wook Choi; Daiki Setoyama; Hyon-Seung Yi
  Endocrinology and metabolism (Seoul, Korea), 37, 6, 891, 900, 2022年12月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, BACKGRUOUND: An excess of thyroid hormones in Graves' disease (GD) has profound effects on systemic energy metabolism that are currently partially understood. In this study, we aimed to provide a comprehensive understanding of the metabolite changes that occur when patients with GD transition from hyperthyroidism to euthyroidism with methimazole treatment. METHODS: Eighteen patients (mean age, 38.6±14.7 years; 66.7% female) with newly diagnosed or relapsed GD attending the endocrinology outpatient clinics in a single institution were recruited between January 2019 and July 2020. All subjects were treated with methimazole to achieve euthyroidism. We explored metabolomics by performing liquid chromatography-mass spectrometry analysis of plasma samples of these patients and then performed multivariate statistical analysis of the metabolomics data. RESULTS: Two hundred metabolites were measured before and after 12 weeks of methimazole treatment in patients with GD. The levels of 61 metabolites, including palmitic acid (C16:0) and oleic acid (C18:1), were elevated in methimazole-naïve patients with GD, and these levels were decreased by methimazole treatment. The levels of another 15 metabolites, including glycine and creatinine, were increased after recovery of euthyroidism upon methimazole treatment in patients with GD. Pathway analysis of metabolomics data showed that hyperthyroidism was closely related to aminoacyl-transfer ribonucleic acid biosynthesis and branched-chain amino acid biosynthesis pathways. CONCLUSION: In this study, significant variations of plasma metabolomic patterns that occur during the transition from hyperthyroidism to euthyroidism were detected in patients with GD via untargeted metabolomics analysis.
• Succinyl-CoA-based energy metabolism dysfunction in chronic heart failure.
  Shingo Takada; Satoshi Maekawa; Takaaki Furihata; Naoya Kakutani; Daiki Setoyama; Koji Ueda; Hideo Nambu; Hikaru Hagiwara; Haruka Handa; Yoshizuki Fumoto; Soichiro Hata; Tomoka Masunaga; Arata Fukushima; Takashi Yokota; Dongchon Kang; Shintaro Kinugawa; Hisataka Sabe
  Proceedings of the National Academy of Sciences of the United States of America, 119, 41, e2203628119, 2022年10月11日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Heart failure (HF) is a leading cause of death and repeated hospitalizations and often involves cardiac mitochondrial dysfunction. However, the underlying mechanisms largely remain elusive. Here, using a mouse model in which myocardial infarction (MI) was induced by coronary artery ligation, we show the metabolic basis of mitochondrial dysfunction in chronic HF. Four weeks after ligation, MI mice showed a significant decrease in myocardial succinyl-CoA levels, and this decrease impaired the mitochondrial oxidative phosphorylation (OXPHOS) capacity. Heme synthesis and ketolysis, and protein levels of several enzymes consuming succinyl-CoA in these events, were increased in MI mice, while enzymes synthesizing succinyl-CoA from α-ketoglutarate and glutamate were also increased. Furthermore, the ADP-specific subunit of succinyl-CoA synthase was reduced, while its GDP-specific subunit was almost unchanged. Administration of 5-aminolevulinic acid, an intermediate in the pathway from succinyl-CoA to heme synthesis, appreciably restored succinyl-CoA levels and OXPHOS capacity and prevented HF progression in MI mice. Previous reports also suggested the presence of succinyl-CoA metabolism abnormalities in cardiac muscles of HF patients. Our results identified that changes in succinyl-CoA usage in different metabolisms of the mitochondrial energy production system is characteristic to chronic HF, and although similar alterations are known to occur in healthy conditions, such as during strenuous exercise, they may often occur irreversibly in chronic HF leading to a decrease in succinyl-CoA. Consequently, nutritional interventions compensating the succinyl-CoA consumption are expected to be promising strategies to treat HF.
• TFAM expression in brown adipocytes confers obesity resistance by secreting extracellular vesicles that promote self-activation.
  Masakazu Fujii; Daiki Setoyama; Kazuhito Gotoh; Yushi Dozono; Mikako Yagi; Masataka Ikeda; Tomomi Ide; Takeshi Uchiumi; Dongchon Kang
  iScience, 25, 9, 104889, 104889, 2022年09月16日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, The occurrence of diet-induced obesity has been increasing worldwide and has become a major health concern. Mitochondria are densely distributed in brown adipose tissue and are involved in lipid consumption. Therefore, increasing energy expenditure through the activation of brown adipocytes may be a potential therapy for obesity. Our findings showed that mitochondrial transcription factor A (TFAM) homozygous transgenic (TgTg) mice had highly activated brown adipocytes and increased expression of oxidative phosphorylation, leading to resistance to obesity. Transplantation models of TFAM-expressing brown adipocytes could mimic the phenotype of TFAM TgTg mice, and proving their anti-obesity effect. We found that brown adipocytes secrete exosomes which enable self-activation in an autocrine and paracrine manner. The secretion was enhanced in TFAM TgTg brown adipocytes, resulting in a higher activation. These findings may lead to a promising treatment strategy for obesity through selective stimulation of exosome secretion.
• Excess intracellular ATP causes neuropathic pain following spinal cord injury.
  Nobuhiko Nakajima; Yuichiro Ohnishi; Masamichi Yamamoto; Daiki Setoyama; Hirohiko Imai; Tomofumi Takenaka; Mari Matsumoto; Koichi Hosomi; Yoichi Saitoh; Hidemasa Furue; Haruhiko Kishima
  Cellular and molecular life sciences : CMLS, 79, 9, 483, 483, 2022年08月16日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Intractable neuropathic pain following spinal cord injury (NP-SCI) reduces a patient's quality of life. Excessive release of ATP into the extracellular space evokes neuroinflammation via purinergic receptor. Neuroinflammation plays an important role in the initiation and maintenance of NP. However, little is known about whether or not extracellular ATP cause NP-SCI. We found in the present study that excess of intracellular ATP at the lesion site evokes at-level NP-SCI. No significant differences in the body weight, locomotor function, or motor behaviors were found in groups that were negative and positive for at-level allodynia. The intracellular ATP level at the lesion site was significantly higher in the allodynia-positive mice than in the allodynia-negative mice. A metabolome analysis revealed that there were no significant differences in the ATP production or degradation between allodynia-negative and allodynia-positive mice. Dorsal horn neurons in allodynia mice were found to be inactivated in the resting state, suggesting that decreased ATP consumption due to neural inactivity leads to a build-up of intracellular ATP. In contrast to the findings in the resting state, mechanical stimulation increased the neural activity of dorsal horn and extracellular ATP release at lesion site. The forced production of intracellular ATP at the lesion site in non-allodynia mice induced allodynia. The inhibition of P2X4 receptors in allodynia mice reduced allodynia. These results suggest that an excess buildup of intracellular ATP in the resting state causes at-level NP-SCI as a result of the extracellular release of ATP with mechanical stimulation.
• Changes in the metabolites of cerebrospinal fluid induced by rTMS in treatment-resistant depression: A pilot study.
  Hiroshi Tateishi; Daiki Setoyama; Takahiro A Kato; Dongchon Kang; Jun Matsushima; Kojiro Nogami; Seiji Mawatari; Ryohei Kojima; Yuka Fujii; Yuta Sakemura; Takumi Shiraishi; Yoshiomi Imamura; Toshihiko Maekawa; Toyoko Asami; Yoshito Mizoguchi; Akira Monji
  Psychiatry research, 313, 114636, 114636, 2022年07月, ［査読有り］, ［筆頭著者］, ［国際誌］
  英語, 研究論文（学術雑誌）, Repetitive transcranial magnetic stimulation (rTMS) improves depressive symptoms in treatment-resistant depression (TRD). This study aimed to analyze changes in cerebrospinal fluid (CSF) metabolites in patients with TRD after rTMS. Five patients with TRD were enrolled in a high frequency (10-Hz) rTMS study. The concentration of 72 CSF metabolites were measured at baseline and at the end of the 6-week rTMS treatment. rTMS significantly increased CSF niacinamide, kynurenine, and creatinine levels and significantly decreased CSF cystine levels, but not the levels of the other 68 CSF metabolites. This is the first CSF metabolomics study on patients with TRD who underwent rTMS.
• Cancer genomic profiling identified dihydropyrimidine dehydrogenase deficiency in bladder cancer promotes sensitivity to gemcitabine.
  Shigehiro Tsukahara; Masaki Shiota; Dai Takamatsu; Shohei Nagakawa; Takashi Matsumoto; Ryo Kiyokoba; Mikako Yagi; Daiki Setoyama; Nozomi Noda; Shinya Matsumoto; Tetsutaro Hayashi; Alberto Contreras-Sanz; Peter C Black; Junichi Inokuchi; Kenichi Kohashi; Yoshinao Oda; Takeshi Uchiumi; Masatoshi Eto; Dongchon Kang
  Scientific reports, 12, 1, 8535, 8535, 2022年05月20日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Chemotherapy is a standard therapy for muscle-invasive bladder cancer (MIBC). However, genomic alterations associated with chemotherapy sensitivity in MIBC have not been fully explored. This study aimed to investigate the genomic landscape of MIBC in association with the response to chemotherapy and to explore the biological role of genomic alterations. Genomic alterations in MIBC were sequenced by targeted exome sequencing of 409 genes. Gene expression in MIBC tissues was analyzed by western blotting, immunohistochemistry, and RNA microarray. Cellular sensitivity to gemcitabine and gemcitabine metabolite was examined in bladder cancer cells after modulation of candidate gene. Targeted exome sequencing in 20 cases with MIBC revealed various genomic alterations including pathogenic missense mutation of DPYD gene encoding dihydropyrimidine dehydrogenase (DPD). Conversely, high DPYD and DPD expression were associated with poor response to gemcitabine-containing chemotherapy among patients with MIBC, as well as gemcitabine resistance in bladder cancer cells. DPD suppression rendered cells sensitive to gemcitabine, while DPD overexpression made cells gemcitabine-resistant through reduced activity of the cytotoxic gemcitabine metabolite difluorodeoxycytidine diphosphate. This study revealed the novel role of DPD in gemcitabine metabolism. It has been suggested that DPYD genomic alterations and DPD expression are potential predictive biomarkers in gemcitabine treatment.
• Mitochondria metabolomics reveals a role of β-nicotinamide mononucleotide metabolism in mitochondrial DNA replication.
  Tomoko Nomiyama; Daiki Setoyama; Takehiro Yasukawa; Dongchon Kang
  Journal of biochemistry, 171, 3, 325, 338, 2022年03月03日, ［査読有り］, ［責任著者］, ［国際誌］
  英語, 研究論文（学術雑誌）, Mitochondrial DNA (mtDNA) replication is tightly regulated and necessary for cellular homeostasis; however, its relationship with mitochondrial metabolism remains unclear. Advances in metabolomics integrated with the rapid isolation of mitochondria will allow for remarkable progress in analyzing mitochondrial metabolism. Here, we propose a novel methodology for mitochondria-targeted metabolomics, which employs a quick isolation procedure using a hemolytic toxin from Streptococcus pyogenes streptolysin O (SLO). SLO isolation of mitochondria from cultured HEK293 cells is time- and labor-saving for simultaneous multi-sample processing and has been applied to various other cell lines in this study. Furthermore, our method can detect the time-dependent reduction in mitochondrial ATP in response to a glycolytic inhibitor 2-deoxyglucose, indicating the suitability to prepare metabolite analysis-competent mitochondria. Using this methodology, we searched for specific mitochondrial metabolites associated with mtDNA replication activation, and nucleotides and NAD+ were identified to be prominently altered. Most notably, treatment of β-nicotinamide mononucleotide (β-NMN), a precursor of NAD+, to HEK293 cells activated and improved the rate of mtDNA replication by increasing nucleotides in mitochondria and decreasing their degradation products: nucleosides. Our results suggest that β-NMN metabolism plays a role in supporting mtDNA replication by maintaining the nucleotide pool balance in the mitochondria.
• Immunometabolic signatures predict recovery from thyrotoxic myopathy in patients with Graves' disease.
  Daiki Setoyama; Ho Yeop Lee; Ji Sun Moon; Jingwen Tian; Yea Eun Kang; Ju Hee Lee; Minho Shong; Dongchon Kang; Hyon-Seung Yi
  Journal of cachexia, sarcopenia and muscle, 13, 1, 355, 367, 2022年02月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, BACKGROUND: Thyroid hormone excess induces protein energy wasting, which in turn promotes muscle weakness and bone loss in patients with Graves' disease. Although most studies have confirmed a relationship between thyrotoxicosis and muscle dysfunction, few have measured changes in plasma metabolites and immune cells during the development and recovery from thyrotoxic myopathy. The aim of this study was to identify specific plasma metabolites and T-cell subsets that predict thyrotoxic myopathy recovery in patients with Graves' disease. METHODS: One hundred patients (mean age, 40.0 ± 14.2 years; 67.0% female), with newly diagnosed or relapsed Graves' disease were enrolled at the start of methimazole treatment. Handgrip strength and Five Times Sit to Stand Test performance time were measured at Weeks 0, 12, and 24. In an additional 35 patients (mean age, 38.9 ± 13.5 years; 65.7% female), plasma metabolites and immunophenotypes of peripheral blood were evaluated at Weeks 0 and 12, and the results of a short physical performance battery assessment were recorded at the same time. RESULTS: In both patient groups, methimazole-induced euthyroidism was associated with improved handgrip strength and lower limb muscle function at 12 weeks. Elevated plasma metabolites including acylcarnitines were restored to normal levels at Week 12 regardless of gender, body mass index, or age (P trend <0.01). Senescent CD8+ CD28- CD57+ T-cell levels in peripheral blood were positively correlated with acylcarnitine levels (P < 0.05) and decreased during thyrotoxicosis recovery (P < 0.05). High levels of senescent CD8+ T cells at Week 0 were significantly associated with small increases in handgrip strength after 12 weeks of methimazole treatment (P < 0.05), but not statistically associated with Five Times Sit to Stand Test performance. CONCLUSIONS: Restoring euthyroidism in Graves' disease patients was associated with improved skeletal muscle function and performance, while thyroid hormone-associated changes in plasma acylcarnitines levels correlated with muscle dysfunction recovery. T-cell senescence-related systemic inflammation correlated with plasma acylcarnitine levels and was also associated with small increases in handgrip strength.
• Fully Automated Quantitative Measurement of Serum Organic Acids via LC-MS/MS for the Diagnosis of Organic Acidemias: Establishment of an Automation System and a Proof-of-Concept Validation.
  Yasushi Ueyanagi; Daiki Setoyama; Daisuke Kawakami; Yuichi Mushimoto; Shinya Matsumoto; Taeko Hotta; Dongchon Kang
  Diagnostics (Basel, Switzerland), 11, 12, 2021年11月25日, ［査読有り］, ［責任著者］, ［国際誌］
  英語, 研究論文（学術雑誌）, Gas chromatography-mass spectrometry has been widely used to analyze hundreds of organic acids in urine to provide a diagnostic basis for organic acidemia. However, it is difficult to operate in clinical laboratories on a daily basis due to sample pretreatment processing. Therefore, we aimed to develop a fully automated system for quantifying serum organic acids using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The pretreatment CLAM-2030 device was connected to an LC-MS/MS system for processing serum under optimized conditions, which included derivatizing serum organic acids using 3-Nitrophenylhydrazine. The derivatized organic acids were separated on a reverse-phase Sceptor HD-C column and detected using negative-ion electrospray ionization multiple reaction monitoring MS. The automated pretreatment-LC-MS/MS system processed serum in less than 1 h and analyzed 19 serum organic acids, which are used to detect organic acidemias. The system exhibited high quantitative sensitivity ranging from approximately 2 to 100 µM with a measurement reproducibility of 10.4% CV. Moreover, a proof-of-concept validation of the system was performed using sera from patients with propionic acidemia (n = 5), methylmalonic acidemia (n = 2), and 3-methylcrotonylglycinuria (n = 1). The levels of marker organic acids specific to each disease were significantly elevated in the sera of the patients compared to those in control samples. The automated pretreatment-LC-MS/MS system can be used as a rapid in-hospital system to measure organic acid levels in serum for the diagnosis of organic acidemias.
• Mitochondrial Lon protease is a gatekeeper for proteins newly imported into the matrix.
  Yuichi Matsushima; Kazuya Takahashi; Song Yue; Yuki Fujiyoshi; Hideaki Yoshioka; Masamune Aihara; Daiki Setoyama; Takeshi Uchiumi; Satoshi Fukuchi; Dongchon Kang
  Communications biology, 4, 1, 974, 974, 2021年08月16日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Human ATP-dependent Lon protease (LONP1) forms homohexameric, ring-shaped complexes. Depletion of LONP1 causes aggregation of a broad range of proteins in the mitochondrial matrix and decreases the levels of their soluble forms. The ATP hydrolysis activity, but not protease activity, of LONP1 is critical for its chaperone-like anti-aggregation activity. LONP1 forms a complex with the import machinery and an incoming protein, and protein aggregation is linked with matrix protein import. LONP1 also contributes to the degradation of imported, aberrant, unprocessed proteins using its protease activity. Taken together, our results show that LONP1 functions as a gatekeeper for specific proteins imported into the mitochondrial matrix.
• The changes in kynurenine metabolites induced by rTMS in treatment-resistant depression: A pilot study.
  Hiroshi Tateishi; Daiki Setoyama; Dongchon Kang; Jun Matsushima; Ryohei Kojima; Yuka Fujii; Seiji Mawatari; Jun Kikuchi; Yuta Sakemura; Junko Fukuchi; Takumi Shiraishi; Toshihiko Maekawa; Takahiro A Kato; Toyoko Asami; Yoshito Mizoguchi; Akira Monji
  Journal of psychiatric research, 138, 194, 199, 2021年06月, ［査読有り］, ［筆頭著者］, ［国際誌］
  英語, 研究論文（学術雑誌）, BACKGROUND: Repetitive transcranial magnetic stimulation (rTMS) is a noninvasive brain stimulation technique that is considered a valuable and promising technique for improving depressive symptoms in treatment-resistant depression (TRD). However, the exact mechanism by which rTMS ameliorates depressive symptoms remains to be clarified. OBJECTIVE: The aim of the present study was to analyzed the changes in metabolites of patients with TRD in the rTMS treatment, especially focusing on the kynurenine (KYN) pathway. METHODS: Thirteen participants with TRD were enrolled in a high-frequency (10 Hz) rTMS study. Cognitive function, depressive symptoms and the concentration of plasma tryptophan (TRP) metabolites were measured at baseline and at the endpoint of rTMS treatment. RESULTS: rTMS treatment significantly improved depressive symptom scores and some subscales of cognitive dysfunction. The present study has demonstrated that rTMS treatment significantly increased plasma TRP levels and significantly decreased plasma serotonin levels, while plasma KYN and kynurenic acid level as well as KYN/TRP ratio remained unchanged. CONCLUSIONS: This is the first metabolomic study of patients with TRD undergoing rTMS treatment. To validate the present results, it is necessary to increase the number of cases including controls, use a sample of cerebrospinal fluid, and measure blood concentration over time in the course of rTMS treatment.
• Mitochondrial translation deficiency impairs NAD+ -mediated lysosomal acidification.
  Mikako Yagi; Takahiro Toshima; Rie Amamoto; Yura Do; Haruka Hirai; Daiki Setoyama; Dongchon Kang; Takeshi Uchiumi
  The EMBO journal, 40, 8, e105268, 2021年04月15日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Mitochondrial translation dysfunction is associated with neurodegenerative and cardiovascular diseases. Cells eliminate defective mitochondria by the lysosomal machinery via autophagy. The relationship between mitochondrial translation and lysosomal function is unknown. In this study, mitochondrial translation-deficient hearts from p32-knockout mice were found to exhibit enlarged lysosomes containing lipofuscin, suggesting impaired lysosome and autolysosome function. These mice also displayed autophagic abnormalities, such as p62 accumulation and LC3 localization around broken mitochondria. The expression of genes encoding for nicotinamide adenine dinucleotide (NAD+ ) biosynthetic enzymes-Nmnat3 and Nampt-and NAD+ levels were decreased, suggesting that NAD+ is essential for maintaining lysosomal acidification. Conversely, nicotinamide mononucleotide (NMN) administration or Nmnat3 overexpression rescued lysosomal acidification. Nmnat3 gene expression is suppressed by HIF1α, a transcription factor that is stabilized by mitochondrial translation dysfunction, suggesting that HIF1α-Nmnat3-mediated NAD+ production is important for lysosomal function. The glycolytic enzymes GAPDH and PGK1 were found associated with lysosomal vesicles, and NAD+ was required for ATP production around lysosomal vesicles. Thus, we conclude that NAD+ content affected by mitochondrial dysfunction is essential for lysosomal maintenance.
• Plasma acetylcholine and nicotinic acid are correlated with focused preference for photographed females in depressed males: an economic game study.
  Hiroaki Kubo; Daiki Setoyama; Motoki Watabe; Masahiro Ohgidani; Kohei Hayakawa; Nobuki Kuwano; Mina Sato-Kasai; Ryoko Katsuki; Shigenobu Kanba; Dongchon Kang; Takahiro A Kato
  Scientific reports, 11, 1, 2199, 2199, 2021年01月26日, ［査読有り］, ［筆頭著者］, ［国際誌］
  英語, 研究論文（学術雑誌）, Interpersonal difficulties are often observed in major depressive disorder (MDD), while the underlying psychological and biological mechanisms have not yet been elucidated. In the present case-control study, a PC-based trust game was conducted for 38 drug-free MDD patients and 38 healthy controls (HC). In the trust game, participants invested money in a partner (trusting behaviors), and also rated each partner's attractiveness (preference for others). In addition, blood biomarkers including metabolites were measured. Both MDD and HC males exhibited more trusting behaviors compared to females. MDD males' preference for ordinary-attractive partners (lay-person photographs) was lower than HC males, whereas their preference for high-attractive females (fashion-model photographs) was similar levels to HC males. This tendency in MDD males could reflect a "focused (narrowed) preference for females". As for blood biomarker analysis, the levels of 37 metabolites including acetylcholine, AMP, GMP, nicotinic acid and tryptophan were significantly different between two groups. Interestingly, among male participants, acetylcholine and nicotinic acid were negatively correlated with the level of focused preference for photographed females. In sum, we have revealed some behavioral, psychological and biological traits of trusting behaviors and preference for others especially in MDD males. Larger studies should be conducted to validate our preliminary findings.
• Personality classification enhances blood metabolome analysis and biotyping for major depressive disorders: two-species investigation.
  Daiki Setoyama; Atsuo Yoshino; Masahiro Takamura; Go Okada; Masaaki Iwata; Kyohei Tsunetomi; Masahiro Ohgidani; Nobuki Kuwano; Junichiro Yoshimoto; Yasumasa Okamoto; Shigeto Yamawaki; Shigenobu Kanba; Dongchon Kang; Takahiro A Kato
  Journal of affective disorders, 279, 20, 30, 2021年01月15日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, BACKGROUND: The relationship between depression and personality has long been suggested, however, biomarker investigations for depression have mostly overlooked this connection. METHODS: We collected personality traits from 100 drug-free patients with major depressive disorders (MDD) and 100 healthy controls based on the Five-Factor Model (FFM) such as Neuroticism (N) and Extraversion (E), and also obtained 63 plasma metabolites profiles by LCMS-based metabolome analysis. RESULTS: Partitional clustering analysis using the NEO-FFI data classified all subjects into three major clusters. Eighty-six subjects belonging to Cluster 1 (C1: less personality-biased group) constituted half of MDD patients and half of healthy controls. C2 constituted 50 subjects mainly MDD patients (N high + E low), and C3 constituted 64 subjects mainly healthy subjects (N low + E high). Using metabolome information, the machine learning model was optimized to discriminate MDD patients from healthy controls among all subjects and C1, respectively. The performance of the model for all subjects was moderate (AUC = 0. 715), while the performance was extremely improved when limited to C1 (AUC = 0. 907). Tryptophan-pathway plasma metabolites including tryptophan, serotonin and kynurenine were significantly lower in MDD patients especially among C1. We also validated metabolomic findings using a social-defeat mice model of stress-induced depression. LIMITATIONS: A case-control study design and sample size is not large. CONCLUSIONS: Our results suggest that personality classification enhances blood biomarker analysis for MDD patients and further translational investigations should be conducted to clarify the biological relationship between personality traits, stress and depression.
• Rostro-caudal different energy metabolism leading to differences in degeneration in spinal cord injury.
  Yuichiro Ohnishi; Masamichi Yamamoto; Yuki Sugiura; Daiki Setoyama; Haruhiko Kishima
  Brain communications, 3, 2, fcab058, 2021年, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Spinal cord injury gradually spreads away from the epicentre of injury. The rate of degeneration on the rostral side of the injury differs from that on the caudal side. Rostral degeneration is an immediate process, while caudal degeneration is delayed. In this study, we demonstrated that the rostro-caudal differences in energy metabolism led to differences in the spread of degeneration in early thoracic cord injury using in vivo imaging. The blood flow at the rostral side of the injury showed ischaemia-reperfusion, while the caudal side presented stable perfusion. The rostral side had an ATP shortage 20 min after spinal cord injury, while the ATP levels were maintained on the caudal side. Breakdown products of purine nucleotides were accumulated at both sides of injury 18 h after spinal cord injury, but the principal metabolites in the tricarboxylic acid cycle and glycolytic pathway were elevated on the caudal side. Although the low-ATP regions expanded at the rostral side of injury until 24 h after spinal cord injury, the caudal-side ATP levels were preserved. The low-ATP regions on the rostral side showed mitochondrial reactive oxygen species production. Administration of 2-deoxy-d-glucose as a glycolysis inhibitor decreased the caudal ATP levels and expanded the low-ATP regions to the caudal side until 24 h after spinal cord injury. These results suggest that deficits in the glycolytic pathway accelerate the caudal degeneration, while immediate rostral degeneration is exacerbated by oxidative stress in early thoracic cord injury.
• Mitochondrial Reactive Oxygen Species Are Essential for the Development of Psoriatic Inflammation.
  Soichi Mizuguchi; Kazuhito Gotoh; Yuya Nakashima; Daiki Setoyama; Yurie Takata; Shouichi Ohga; Dongchon Kang
  Frontiers in immunology, 12, 714897, 714897, 2021年, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Psoriasis is a common immune-mediated, chronic, inflammatory skin disease that affects approximately 2-3% of the population worldwide. Although there is increasing evidence regarding the essential roles of the interleukin (IL)-23/IL-17 axis and dendritic cell (DC)-T cell crosstalk in the development of skin inflammation, the contributions of mitochondrial function to psoriasis are unclear. In a mouse model of imiquimod (IMQ)-induced psoriasiform skin inflammation, we found that hematopoietic cell-specific genetic deletion of p32/C1qbp, a regulator of mitochondrial protein synthesis and metabolism, protects mice from IMQ-induced psoriatic inflammation. Additionally, we demonstrate that p32/C1qbp is an important regulator of IMQ-induced DC activation, both in vivo and in vitro. We also found that p32/C1qbp-deficient DCs exhibited impaired production of IL-1β, IL-23, and mitochondrial reactive oxygen species (mtROS) after IMQ stimulation. Because the inhibition of mtROS suppressed IMQ-induced DC activation and psoriatic inflammation, we presume that p32/C1qbp and mtROS can serve as therapeutic targets in psoriasis.
• Attenuating Effect of Chlorella Extract on NLRP3 Inflammasome Activation by Mitochondrial Reactive Oxygen Species.
  Yuya Nakashima; Kazuhito Gotoh; Soichi Mizuguchi; Daiki Setoyama; Yurie Takata; Toshihiro Kanno; Dongchon Kang
  Frontiers in nutrition, 8, 763492, 763492, 2021年, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, The NOD-like receptor family, pyrin domain-containing protein 3 (NLRP3) inflammasome has been linked to the pathogenesis of a wide variety of human diseases. Although many drugs and inhibitors have been developed to treat NLRP3-associated diseases, only limited clinical data support their efficacy and safety. Chlorella, a unicellular green alga that is widely and safely used as a food supplement, contains various antioxidants. In this study, we obtained a fat-soluble extract from Chlorella (CE) and demonstrated that it reduced NLRP3 inflammasome activation by inhibiting mitochondrial reactive oxygen species and caspase-1 activation. In addition, CE supplementation attenuated lipopolysaccharide-induced interleukin 1β transcription through activation of hypoxia-inducible factor 1α in vitro and in vivo. As Chlorella is a safe and useful food supplement, it may be a practical pharmacological approach for treating NLRP3-driven diseases.
• Blood metabolic signatures of hikikomori, pathological social withdrawal.
  Daiki Setoyama; Toshio Matsushima; Kohei Hayakawa; Tomohiro Nakao; Shigenobu Kanba; Dongchon Kang; Takahiro A Kato
  Dialogues in clinical neuroscience, 23, 1, 14, 28, 2021年, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, BACKGROUND: A severe form of pathological social withdrawal, 'hikikomori,' has been acknowledged in Japan, spreading worldwide, and becoming a global health issue. The pathophysiology of hikikomori has not been clarified, and its biological traits remain unexplored. METHODS: Drug-free patients with hikikomori (n = 42) and healthy controls (n = 41) were recruited. Psychological assessments for the severity of hikikomori and depression were conducted. Blood biochemical tests and plasma metabolome analysis were performed. Based on the integrated information, machine-learning models were created to discriminate cases of hikikomori from healthy controls, predict hikikomori severity, stratify the cases, and identify metabolic signatures that contribute to each model. RESULTS: Long-chain acylcarnitine levels were remarkably higher in patients with hikikomori; bilirubin, arginine, ornithine, and serum arginase were significantly different in male patients with hikikomori. The discriminative random forest model was highly performant, exhibiting an area under the ROC curve of 0.854 (confidential interval = 0.648-1.000). To predict hikikomori severity, a partial least squares PLS-regression model was successfully created with high linearity and practical accuracy. In addition, blood serum uric acid and plasma cholesterol esters contributed to the stratification of cases. CONCLUSIONS: These findings reveal the blood metabolic signatures of hikikomori, which are key to elucidating the pathophysiology of hikikomori and also useful as an index for monitoring the treatment course for rehabilitation.
• Beta-hydroxybutyrate, an endogenous NLRP3 inflammasome inhibitor, attenuates anxiety-related behavior in a rodent post-traumatic stress disorder model.
  Takehiko Yamanashi; Masaaki Iwata; Midori Shibushita; Kyohei Tsunetomi; Mayu Nagata; Naofumi Kajitani; Akihiko Miura; Ryoichi Matsuo; Tsuyoshi Nishiguchi; Takahiro A Kato; Daiki Setoyama; Yukihiko Shirayama; Ken Watanabe; Gen Shinozaki; Koichi Kaneko
  Scientific reports, 10, 1, 21629, 21629, 2020年12月10日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Accumulating evidence suggests that elevated inflammation contributes to the pathophysiology of post-traumatic stress disorder (PTSD) and that anti-inflammatory drugs might be a new treatment strategy for PTSD. It has been reported that beta-hydroxybutyrate (BHB), one of the main ketone bodies produced, can have an anti-inflammatory and antidepressant effect. Here, we investigated the potential anti-anxiety and anti-inflammatory effects of BHB using a rodent PTSD model, induced by single prolonged stress (SPS). Male, Sprague-Dawley rats were employed in this study. Repeated administration of BHB attenuated SPS-induced anxiety-related behaviors evaluated by the elevated plus maze test. SPS increased the serum levels of TNF-α and IL-1β. In contrast, BHB administration partially attenuated the increase of serum TNF-α. These findings demonstrate that BHB exerts its anxiolytic effects, possibly by inhibiting systemic TNF-α. Hence, BHB may be a novel therapeutic candidate for the treatment of PTSD.
• GNAO1 organizes the cytoskeletal remodeling and firing of developing neurons.
  Satoshi Akamine; Sayaka Okuzono; Hiroyuki Yamamoto; Daiki Setoyama; Noriaki Sagata; Masahiro Ohgidani; Takahiro A Kato; Tohru Ishitani; Hiroki Kato; Keiji Masuda; Yuki Matsushita; Hiroaki Ono; Yoshito Ishizaki; Masafumi Sanefuji; Hirotomo Saitsu; Naomichi Matsumoto; Dongchon Kang; Shigenobu Kanba; Yusaku Nakabeppu; Yasunari Sakai; Shouichi Ohga
  FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 34, 12, 16601, 16621, 2020年12月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Developmental and epileptic encephalopathy (DEE) represents a group of neurodevelopmental disorders characterized by infantile-onset intractable seizures and unfavorable prognosis of psychomotor development. To date, hundreds of genes have been linked to the onset of DEE. GNAO1 is a DEE-associated gene encoding the alpha-O1 subunit of guanine nucleotide-binding protein (GαO ). Despite the increasing number of reported children with GNAO1 encephalopathy, the molecular mechanisms underlying their neurodevelopmental phenotypes remain elusive. We herein present that co-immunoprecipitation and mass spectrometry analyses identified another DEE-associated protein, SPTAN1, as an interacting partner of GαO . Silencing of endogenous Gnao1 attenuated the neurite outgrowth and calcium-dependent signaling. Inactivation of GNAO1 in human-induced pluripotent stem cells gave rise to anomalous brain organoids that only weakly expressed SPTAN1 and Ankyrin-G. Furthermore, GNAO1-deficient organoids failed to conduct synchronized firing to adjacent neurons. These data indicate that GαO and other DEE-associated proteins organize the cytoskeletal remodeling and functional polarity of neurons in the developing brain.
• Mitochondrial Protein Synthesis Is Essential for Terminal Differentiation of CD45- TER119-Erythroid and Lymphoid Progenitors.
  Kazuhito Gotoh; Yuya Kunisaki; Soichi Mizuguchi; Daiki Setoyama; Kentaro Hosokawa; Hisayuki Yao; Yuya Nakashima; Mikako Yagi; Takeshi Uchiumi; Yuichiro Semba; Jumpei Nogami; Koichi Akashi; Fumio Arai; Dongchon Kang
  iScience, 23, 11, 101654, 101654, 2020年11月20日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, p32/C1qbp regulates mitochondrial protein synthesis and is essential for oxidative phosphorylation in mitochondria. Although dysfunction of p32/C1qbp impairs fetal development and immune responses, its role in hematopoietic differentiation remains unclear. Here, we found that mitochondrial dysfunction affected terminal differentiation of newly identified erythroid/B-lymphoid progenitors among CD45- Ter119- CD31- triple-negative cells (TNCs) in bone marrow. Hematopoietic cell-specific genetic deletion of p32/C1qbp (p32cKO) in mice caused anemia and B-lymphopenia without reduction of hematopoietic stem/progenitor cells. In addition, p32cKO mice were susceptible to hematopoietic stress with delayed recovery from anemia. p32/C1qbp-deficient CD51- TNCs exhibited impaired mitochondrial oxidation that consequently led to inactivation of mTORC1 signaling, which is essential for erythropoiesis. These findings uncover the importance of mitochondria, especially at the stage of TNCs during erythropoiesis, suggesting that dysregulation of mitochondrial protein synthesis is a cause of anemia and B-lymphopenia with an unknown pathology.
• Kynurenic acid is a potential overlapped biomarker between diagnosis and treatment response for depression from metabolome analysis.
  Hisayuki Erabi; Go Okada; Chiyo Shibasaki; Daiki Setoyama; Dongchon Kang; Masahiro Takamura; Atsuo Yoshino; Manabu Fuchikami; Akiko Kurata; Takahiro A Kato; Shigeto Yamawaki; Yasumasa Okamoto
  Scientific reports, 10, 1, 16822, 16822, 2020年10月08日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Since optimal treatment at an early stage leads to remission of symptoms and recovery of function, putative biomarkers leading to early diagnosis and prediction of therapeutic responses are desired. The current study aimed to use a metabolomic approach to extract metabolites involved in both the diagnosis of major depressive disorder (MDD) and the prediction of therapeutic response for escitalopram. We compared plasma metabolites of MDD patients (n = 88) with those in healthy participants (n = 88) and found significant differences in the concentrations of 20 metabolites. We measured the Hamilton Rating Scale for Depression (HRSD) on 62 patients who completed approximately six-week treatment with escitalopram before and after treatment and found that kynurenic acid and kynurenine were significantly and negatively associated with HRSD reduction. Only one metabolite, kynurenic acid, was detected among 73 metabolites for overlapped biomarkers. Kynurenic acid was lower in MDD, and lower levels showed a better therapeutic response to escitalopram. Kynurenic acid is a metabolite in the kynurenine pathway that has been widely accepted as being a major mechanism in MDD. Overlapping biomarkers that facilitate diagnosis and prediction of the treatment response may help to improve disease classification and reduce the exposure of patients to less effective treatments in MDD.
• Characterization and function of medium and large extracellular vesicles from plasma and urine by surface antigens and Annexin V
  Igami K.; Uchiumi T.; Ueda S.; Kamioka K.; Setoyama D.; Gotoh K.
  PeerJ Analytical Chemistry. 2020;2:e4., 2020年, ［査読有り］
• Igami K, Uchiumi T, Ueda S, Kamioka K, Setoyama D, Gotoh K,
  Characterization; function of medium; large extracellular vesicles from plasma; urine by surface antigens; Annexin V.
  PeerJ Analytical Chemistry., 2020年, ［査読有り］
• Linoleic acid improves assembly of the CII subunit and CIII2/CIV complex of the mitochondrial oxidative phosphorylation system in heart failure.
  Satoshi Maekawa; Shingo Takada; Hideo Nambu; Takaaki Furihata; Naoya Kakutani; Daiki Setoyama; Yasushi Ueyanagi; Dongchon Kang; Hisataka Sabe; Shintaro Kinugawa
  Cell communication and signaling : CCS, 17, 1, 128, 128, 2019年10月16日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, BACKGROUND: Linoleic acid is the major fatty acid moiety of cardiolipin, which is central to the assembly of components involved in mitochondrial oxidative phosphorylation (OXPHOS). Although linoleic acid is an essential nutrient, its excess intake is harmful to health. On the other hand, linoleic acid has been shown to prevent the reduction in cardiolipin content and to improve mitochondrial function in aged rats with spontaneous hypertensive heart failure (HF). In this study, we found that lower dietary intake of linoleic acid in HF patients statistically correlates with greater severity of HF, and we investigated the mechanisms therein involved. METHODS: HF patients, who were classified as New York Heart Association (NYHA) functional class I (n = 45), II (n = 93), and III (n = 15), were analyzed regarding their dietary intakes of different fatty acids during the one month prior to the study. Then, using a mouse model of HF, we confirmed reduced cardiolipin levels in their cardiac myocytes, and then analyzed the mechanisms by which dietary supplementation of linoleic acid improves cardiac malfunction of mitochondria. RESULTS: The dietary intake of linoleic acid was significantly lower in NYHA III patients, as compared to NYHA II patients. In HF model mice, both CI-based and CII-based OXPHOS activities were affected together with reduced cardiolipin levels. Silencing of CRLS1, which encodes cardiolipin synthetase, in cultured cardiomyocytes phenocopied these events. Feeding HF mice with linoleic acid improved both CI-based and CII-based respiration as well as left ventricular function, together with an increase in cardiolipin levels. However, although assembly of the respirasome (i.e., CI/CIII2/CIV complex), as well as assembly of CII subunits and the CIII2/CIV complex statistically correlated with cardiolipin levels in cultured cardiomyocytes, respirasome assembly was not notably restored by dietary linoleic acid in HF mice. Therefore, although linoleic acid may significantly improve both CI-based and CII-based respiration of cardiomyocytes, respirasomes impaired by HF were not easily repaired by the dietary intake of linoleic acid. CONCLUSIONS: Dietary supplement of linoleic acid is beneficial for improving cardiac malfunction in HF, but is unable to completely cure HF.
• Impaired plasmalogen synthesis dysregulates liver X receptor-dependent transcription in cerebellum.
  Masanori Honsho; Fabian Dorninger; Yuichi Abe; Daiki Setoyama; Ryohei Ohgi; Takeshi Uchiumi; Dongchon Kang; Johannes Berger; Yukio Fujiki
  Journal of biochemistry, 166, 4, 353, 361, 2019年10月01日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Synthesis of ethanolamine plasmalogen (PlsEtn) is regulated by modulating the stability of fatty acyl-CoA reductase 1 (Far1) on peroxisomal membrane, a rate-limiting enzyme in plasmalogen synthesis. Dysregulation of plasmalogen homeostasis impairs cholesterol biosynthesis in cultured cells by altering the stability of squalene epoxidase (SQLE). However, regulation of PlsEtn synthesis and physiological consequences of plasmalogen homeostasis in tissues remain unknown. In the present study, we found that the protein but not the transcription level of Far1 in the cerebellum of the Pex14 mutant mouse expressing Pex14p lacking its C-terminal region (Pex14ΔC/ΔC) is higher than that from wild-type mouse, suggesting that Far1 is stabilized by the lowered level of PlsEtn. The protein level of SQLE was increased, whereas the transcriptional activity of the liver X receptors (LXRs), ligand-activated transcription factors of the nuclear receptor superfamily, is lowered in the cerebellum of Pex14ΔC/ΔC and the mice deficient in dihydroxyacetonephosphate acyltransferase, the initial enzyme for the synthesis of PlsEtn. These results suggest that the reduction of plasmalogens in the cerebellum more likely compromises the cholesterol homeostasis, thereby reducing the transcriptional activities of LXRs, master regulators of cholesterol homeostasis.
• Development and validation of the 22-item Tarumi's Modern-Type Depression Trait Scale: Avoidance of Social Roles, Complaint, and Low Self-Esteem (TACS-22).
  Takahiro A Kato; Ryoko Katsuki; Hiroaki Kubo; Norihiro Shimokawa; Mina Sato-Kasai; Kohei Hayakawa; Nobuki Kuwano; Wakako Umene-Nakano; Masaru Tateno; Daiki Setoyama; Dongchon Kang; Motoki Watabe; Shinji Sakamoto; Alan R Teo; Shigenobu Kanba
  Psychiatry and clinical neurosciences, 73, 8, 448, 457, 2019年08月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, AIM: Understanding premorbid personality is important, especially when considering treatment selection. Historically, the premorbid personality of patients with major depression in Japan was described as Shuchaku-kishitsu [similar to Typus melancholicus], as proposed by Shimoda in the 1930s. Since around 2000, there have been increased reports in Japan of young adults with depression who have had premorbid personality differing from the traditional type. In 2005, Tarumi termed this novel condition 'dysthymic-type depression,' and more recently the condition has been called Shin-gata/Gendai-gata Utsu-byo [modern-type depression (MTD)]. We recently developed a semi-structured diagnostic interview to evaluate MTD. Development of a tool that enables understanding of premorbid personality in a short time, especially at the early stage of treatment, is desirable. The object of this study was to develop a self-report scale to evaluate the traits of MTD, and to assess the scale's psychometric properties, diagnostic accuracy, and biological validity. METHODS: A sample of 340 participants from clinical and community settings completed measures. Psychometric properties were assessed with factor analysis. Diagnostic accuracy of the MTD traits was compared against a semi-structured interview. RESULTS: The questionnaire contained 22 items across three subscales, thus we termed it the 22-item Tarumi's Modern-Type Depression Trait Scale: Avoidance of Social Roles, Complaint, and Low Self-Esteem (TACS-22). Internal consistency, test-retest reliability, and convergent validity were all satisfactory. Among patients with major depression, the area under the curve was 0.757 (sensitivity of 63.1% and specificity of 82.9%) and the score was positively correlated with plasma tryptophan. CONCLUSION: The TACS-22 possessed adequate psychometric properties and diagnostic accuracy in an initial sample of Japanese adults. Additional research on its ability to support clinical assessment of MTD is warranted.
• Suicide and Microglia: Recent Findings and Future Perspectives Based on Human Studies.
  Hisaomi Suzuki; Masahiro Ohgidani; Nobuki Kuwano; Fabrice Chrétien; Geoffroy Lorin de la Grandmaison; Mitsumoto Onaya; Itaru Tominaga; Daiki Setoyama; Dongchon Kang; Masaru Mimura; Shigenobu Kanba; Takahiro A Kato
  Frontiers in cellular neuroscience, 13, 31, 31, 2019年, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Suicide is one of the most disastrous outcomes for psychiatric disorders. Recent advances in biological psychiatry have suggested a positive relationship between some specific brain abnormalities and specific symptoms in psychiatric disorders whose organic bases were previously completely unknown. Microglia, immune cells in the brain, are regarded to play crucial roles in brain inflammation by releasing inflammatory mediators and are suggested to contribute to various psychiatric disorders such as depression and schizophrenia. Recently, activated microglia have been suggested to be one of the possible contributing cells to suicide and suicidal behaviors via various mechanisms especially including the tryptophan-kynurenine pathway. Animal model research focusing on psychiatric disorders has a long history, however, there are only limited animal models that can properly express psychiatric symptoms. In particular, to our knowledge, animal models of human suicidal behaviors have not been established. Suicide is believed to be limited to humans, therefore human subjects should be the targets of research despite various ethical and technical limitations. From this perspective, we introduce human biological studies focusing on suicide and microglia. We first present neuropathological studies using the human postmortem brain of suicide victims. Second, we show recent findings based on positron emission tomography (PET) imaging and peripheral blood biomarker analysis on living subjects with suicidal ideation and/or suicide-related behaviors especially focusing on the tryptophan-kynurenine pathway. Finally, we propose future perspectives and tasks to clarify the role of microglia in suicide using multi-dimensional analytical methods focusing on human subjects with suicidal ideation, suicide-related behaviors and suicide victims.
• Mitochondrial p32/C1qbp Is a Critical Regulator of Dendritic Cell Metabolism and Maturation.
  Kazuhito Gotoh; Takafumi Morisaki; Daiki Setoyama; Katsuhiko Sasaki; Mikako Yagi; Ko Igami; Soichi Mizuguchi; Takeshi Uchiumi; Yoshinori Fukui; Dongchon Kang
  Cell reports, 25, 7, 1800, 1815, 2018年11月13日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Dendritic cell (DC) maturation induced by Toll-like receptor agonists requires activation of downstream signal transduction and metabolic changes. The endogenous metabolite citrate has recently emerged as a modulator of DC activation. However, the metabolic requirements that support citrate production remain poorly defined. Here, we demonstrate that p32/C1qbp, which functions as a multifunctional chaperone protein in mitochondria, supports mitochondrial metabolism and DC maturation. Metabolic analysis revealed that the citrate increase induced by lipopolysaccharide (LPS) is impaired in p32-deficient DCs. We also found that p32 interacts with dihydrolipoamide S-acetyltransferase (E2 component of pyruvate dehydrogenase [PDH] complex) and positively regulates PDH activity in DCs. Therefore, we suggest that DC maturation is regulated by citrate production via p32-dependent PDH activity. p32-null mice administered a PDH inhibitor show decreased DC maturation and ovalbumin-specific IgG production in vivo, suggesting that p32 may serve as a therapeutic target for DC-related autoimmune diseases.
• Tryptophan-kynurenine and lipid related metabolites as blood biomarkers for first-episode drug-naïve patients with major depressive disorder: An exploratory pilot case-control study.
  Nobuki Kuwano; Takahiro A Kato; Daiki Setoyama; Mina Sato-Kasai; Norihiro Shimokawa; Kohei Hayakawa; Masahiro Ohgidani; Noriaki Sagata; Hiroaki Kubo; Junji Kishimoto; Dongchon Kang; Shigenob Kanba
  Journal of affective disorders, 231, 74, 82, 2018年04月15日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, BACKGROUND: Early intervention in depression has been critical to prevent its negative impact including suicide. Recent blood biomarker studies for major depressive disorder (MDD) have suggested that tryptophan-kynurenine and lipid related metabolites are involved in the pathophysiology of MDD. However, there have been limited studies investigating these blood biomarkers in first-episode drug-naïve MDD, which are particularly important for early intervention in depression. METHODS: As an exploratory pilot case-control study, we examined the above blood biomarkers, and analyzed how these biomarkers are associated with clinical variables in first-episode drug-naïve MDD patients, based on metabolome/lipidome analysis. RESULTS: Plasma tryptophan and kynurenine levels were significantly lower in MDD group (N = 15) compared to healthy controls (HC) group (N = 19), and plasma tryptophan was the significant biomarker to identify MDD group (area under the curve = 0.740). Lower serum high density lipoprotein-cholesterol (HDL-C) was the predictive biomarker for severity of depression in MDD group (R2 = 0.444). Interestingly, depressive symptoms were variously correlated with plasma tryptophan-kynurenine and lipid related metabolites. Moreover, plasma tryptophan-kynurenine metabolites and cholesteryl esters (CEs) were significantly correlated in MDD group, but not in HC group. LIMITATIONS: This study had small sample size, and we did not use the multiple test correction. CONCLUSIONS: This is the first study to suggest that not only tryptophan-kynurenine metabolites but also HDL-C and CEs are important blood biomarkers for first-episode drug-naïve MDD patients. The present study sheds new light on early intervention in clinical practice in depression, and further clinical studies especially large-scale prospective studies are warranted.
• Analysis of spatiotemporal metabolomic dynamics for sensitively monitoring biological alterations in cisplatin-induced acute kidney injury.
  Miho Irie; Eisuke Hayakawa; Yoshinori Fujimura; Youhei Honda; Daiki Setoyama; Hiroyuki Wariishi; Fuminori Hyodo; Daisuke Miura
  Biochemical and biophysical research communications, 496, 1, 140, 146, 2018年01月29日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Clinical application of the major anticancer drug, cisplatin, is limited by severe side effects, especially acute kidney injury (AKI) caused by nephrotoxicity. The detailed metabolic mechanism is still largely unknown. Here, we used an integrated technique combining mass spectrometry imaging (MSI) and liquid chromatography-mass spectrometry (LC-MS) to visualize the diverse spatiotemporal metabolic dynamics in the mouse kidney after cisplatin dosing. Biological responses to cisplatin was more sensitively detected within 24 h as a metabolic alteration, which is much earlier than possible with the conventional clinical chemistry method of blood urea nitrogen (BUN) measurement. Region-specific changes (e.g., medulla and cortex) in metabolites related to DNA damage and energy generation were observed over the 72-h exposure period. Therefore, this metabolomics approach may become a novel strategy for elucidating early renal responses to cisplatin, prior to the detection of kidney damage evaluated by conventional method.
• Doxycycline induces apoptosis via ER stress selectively to cells with a cancer stem cell-like properties: importance of stem cell plasticity.
  Takashi Matsumoto; Takeshi Uchiumi; Keisuke Monji; Mikako Yagi; Daiki Setoyama; Rie Amamoto; Yuichi Matsushima; Masaki Shiota; Masatoshi Eto; Dongchon Kang
  Oncogenesis, 6, 11, 397, 397, 2017年11月29日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Tumor heterogeneity can be traced back to a small subset of cancer stem cells (CSCs), which can be derived from a single stem cell and show chemoresistance. Recent studies showed that CSCs are sensitive to mitochondrial targeting antibiotics such as doxycycline. However, little is known about how cancer cells undergo sphere formation and how antibiotics inhibit CSC proliferation. Here we show that under sphere-forming assay conditions, prostate cancer cells acquired CSC-like properties: promoted mitochondrial respiratory chain activity, expression of characteristic CSC markers and resistance to anticancer agents. Furthermore, those CSC-like properties could reversibly change depending on the culture conditions, suggesting some kinds of CSCs have plasticity in tumor microenvironments. The sphere-forming cells (i.e. cancer stem-like cells) showed increased contact between mitochondria and mitochondrial associated-endoplasmic reticulum (ER) membranes (MAM). Mitochondrial targeting doxycycline induced activating transcription factor 4 (ATF4) mediated expression of ER stress response and led to p53-upregulated modulator of apoptosis (PUMA)-dependent apoptosis only in the cancer stem-like cells. We also found that doxycycline effectively suppressed the sphere formation in vitro and blocked CD44v9-expressing tumor growth in vivo. In summary, these data provide new molecular findings that monolayer cancer cells acquire CSC-like properties in a reversible manner. These findings provide important insights into CSC biology and a potential new treatment of targeting mitochondria dependency.
• Neural-specific deletion of mitochondrial p32/C1qbp leads to leukoencephalopathy due to undifferentiated oligodendrocyte and axon degeneration.
  Mikako Yagi; Takeshi Uchiumi; Noriaki Sagata; Daiki Setoyama; Rie Amamoto; Yuichi Matsushima; Dongchon Kang
  Scientific reports, 7, 1, 15131, 15131, 2017年11月09日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Mitochondrial dysfunction is a critical step in the pathogenesis of many neurodegenerative diseases. The p32/ C1qbp gene functions as an essential RNA and protein chaperone in mitochondrial translation, and is indispensable for embryonic development. However, little is known about the consequences of mitochondrial dysfunction of p32 deletion in the brain development. Here, we found that mice lacking p32 in the central nervous system (p32cKO mice) showed white matter degeneration accompanied by progressive oligodendrocyte loss, axon degeneration and vacuolation in the mid brain and brain stem regions. Furthermore, p32cKO mice died within 8 weeks of birth. We also found that p32-deficient oligodendrocytes and neurons showed reduced oligodendrocyte differentiation and axon degeneration in primary culture. We show that mitochondrial disruption activates an adaptive program known as the integrated stress response (ISR). Mitochondrial respiratory chain function in oligodendrocytes and neurons is, therefore, essential for myelination and axon maintenance, respectively, suggesting that mitochondrial respiratory chain dysfunction in the central nervous system contributes to leukoencephalopathy.
• Dysregulated gene expressions of MEX3D, FOS and BCL2 in human induced-neuronal (iN) cells from NF1 patients: a pilot study.
  Noriaki Sagata; Takahiro A Kato; Shin-Ichi Kano; Masahiro Ohgidani; Norihiro Shimokawa; Mina Sato-Kasai; Kohei Hayakawa; Nobuki Kuwano; Ashley M Wilson; Koko Ishizuka; Shiori Kato; Takeshi Nakahara; Makiko Nakahara-Kido; Daiki Setoyama; Yasunari Sakai; Shouichi Ohga; Masutaka Furue; Akira Sawa; Shigenobu Kanba
  Scientific reports, 7, 1, 13905, 13905, 2017年10月24日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Direct conversion technique to produce induced-neuronal (iN) cells from human fibroblasts within 2 weeks is expected to discover unknown neuronal phenotypes of neuropsychiatric disorders. Here, we present unique gene expression profiles in iN cells from patients with neurofibromatosis type 1 (NF1), a single-gene multifaceted disorder with comparatively high co-occurrence of autism spectrum disorder (ASD). Microarray-based transcriptomic analysis on iN cells from male healthy controls and male NF1 patients (NF1-iN cells) revealed that 149 genes expressions were significantly different (110 upregulated and 39 downregulated). We validated that mRNA of MEX3D (mex-3 RNA binding family member D) was lower in NF1-iN cells by real-time PCR with 12 sex-mixed samples. In NF1-iN cells on day 14, higher expression of FOS mRNA was observed with lower expression of MEX3D mRNA. Interestingly, BCL2 mRNA was higher in NF1-iN cells on day 5 (early-period) but not on day 14. Our data suggest that aberrant molecular signals due to NF1 mutations may disturb gene expressions, a subset of which defines continuum of the neuronal phenotypes of NF1 with ASD. Further translational studies using induced pluripotent stem (iPS) cell-derived neuronal cells are needed to validate our preliminary findings especially confirming meanings of analysis using early-period iN cells.
• Cardiomyocyte-specific loss of mitochondrial p32/C1qbp causes cardiomyopathy and activates stress responses.
  Toshiro Saito; Takeshi Uchiumi; Mikako Yagi; Rie Amamoto; Daiki Setoyama; Yuichi Matsushima; Dongchon Kang
  Cardiovascular research, 113, 10, 1173, 1185, 2017年08月01日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, AIMS: Mitochondria are important organelles, dedicated to energy production. Mitochondrial p32/C1qbp, which functions as an RNA and protein chaperone, interacts with mitochondrial mRNA and is indispensable for mitochondrial function through its regulation of mitochondrial translation in cultured cell lines. However, the precise role of p32/C1qbp in vivo is poorly understood because of embryonic lethality in the systemic p32-deficient mouse. The goal of this study was to examine the physiological function of mitochondrial p32/C1qbp in the heart. METHODS AND RESULTS: We investigated the role of p32 in regulating cardiac function in mice using a Cre-loxP recombinase technology against p32 with tamoxifen-inducible knockdown or genetic ablation during postnatal periods. Cardiomyocyte-specific deletion of p32 resulted in contractile dysfunction, cardiac dilatation and cardiac fibrosis, compared with hearts of control mice. We also found decreased COX1 expression, decreased rates of oxygen consumption and increased oxidative stress, indicating that these mice had cardiac mitochondrial dysfunction provoked by p32-deficiency at early stage. Next, we investigated lifespan in cardiac-specific p32-deficient mice. The mice died beginning at 12 months and their median lifespan was ∼14 months. Cardiac mitochondria in the p32-deficient mice showed disordered alignment, enlargement and abnormalities in their internal structure by electron microscopy. We observed that, in p32-deficient compared with control myocytes, AMPKɑ was constitutively phosphorylated and 4EBP-1 and ribosomal S6K were less phosphorylated, suggesting impairment of mammalian target of rapamycin signalling. Finally, we found that expression levels of mitokines such as FGF21 and of integrated stress response genes were significantly increased. Metabolic analysis demonstrated that the urea cycle was impaired in the p32-deficient hearts. CONCLUSION: These findings support a key role for mitochondrial p32 protein in cardiac myocytes modulating mitochondrial translation and function, and thereby survival.
• p32 is Required for Appropriate Interleukin-6 Production Upon LPS Stimulation and Protects Mice from Endotoxin Shock.
  Katsuhiko Sasaki; Kazuhito Gotoh; Sho Miake; Daiki Setoyama; Mikako Yagi; Ko Igami; Takeshi Uchiumi; Dongchon Kang
  EBioMedicine, 20, 161, 172, 2017年06月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Sepsis is a major cause of morbidity and mortality in seriously ill patients and mitochondrial dysfunction is associated with poor outcomes in septic patients. Although interleukin-6 (IL-6) is a good prognostic marker for sepsis, the relationship between mitochondrial dysfunction and IL-6 remains poorly understood. We identified p32/C1QBP/HABP1 as a regulator of IL-6 production in response to lipopolysaccharide (LPS). LPS induced IL-6 overproduction in p32 deficient mouse embryonic fibroblasts (MEFs) through NF-κB independent but activating transcription factor (ATF) 4 dependent pathways. Short hairpin RNA-based knockdown of ATF4 in p32 deficient MEFs markedly inhibited LPS-induced IL-6 production. Furthermore, MEFs treated with chloramphenicol, an inhibitor of mitochondrial translation, produced excessive IL-6 via ATF4 pathways. Using a LPS-induced endotoxin shock model, mice with p32 ablation in myeloid cells showed increased lethality and overproduction of IL-6. Thus, this study provides a molecular link how mitochondrial dysfunction leads to IL-6 overproduction and poor prognosis of sepsis.
• Targeting Ras-Driven Cancer Cell Survival and Invasion through Selective Inhibition of DOCK1.
  Hirotada Tajiri; Takehito Uruno; Takahiro Shirai; Daisuke Takaya; Shigeki Matsunaga; Daiki Setoyama; Mayuki Watanabe; Mutsuko Kukimoto-Niino; Kounosuke Oisaki; Miho Ushijima; Fumiyuki Sanematsu; Teruki Honma; Takaho Terada; Eiji Oki; Senji Shirasawa; Yoshihiko Maehara; Dongchon Kang; Jean-François Côté; Shigeyuki Yokoyama; Motomu Kanai; Yoshinori Fukui
  Cell reports, 19, 5, 969, 980, 2017年05月02日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Oncogenic Ras plays a key role in cancer initiation but also contributes to malignant phenotypes by stimulating nutrient uptake and promoting invasive migration. Because these latter cellular responses require Rac-mediated remodeling of the actin cytoskeleton, we hypothesized that molecules involved in Rac activation may be valuable targets for cancer therapy. We report that genetic inactivation of the Rac-specific guanine nucleotide exchange factor DOCK1 ablates both macropinocytosis-dependent nutrient uptake and cellular invasion in Ras-transformed cells. By screening chemical libraries, we have identified 1-(2-(3'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)-2-oxoethyl)-5-pyrrolidinylsulfonyl-2(1H)-pyridone (TBOPP) as a selective inhibitor of DOCK1. TBOPP dampened DOCK1-mediated invasion, macropinocytosis, and survival under the condition of glutamine deprivation without impairing the biological functions of the closely related DOCK2 and DOCK5 proteins. Furthermore, TBOPP treatment suppressed cancer metastasis and growth in vivo in mice. Our results demonstrate that selective pharmacological inhibition of DOCK1 could be a therapeutic approach to target cancer cell survival and invasion.
• Serum depletion induced cancer stem cell-like phenotype due to nitric oxide synthesis in oncogenic HRas transformed cells.
  Keisuke Monji; Takeshi Uchiumi; Saki Hoshizawa; Mikako Yagi; Takashi Matsumoto; Daiki Setoyama; Yuichi Matsushima; Kazuhito Gotoh; Rie Amamoto; Donchon Kang
  Oncotarget, 7, 46, 75221, 75234, 2016年11月15日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Cancer cells rewire their metabolism and mitochondrial oxidative phosphorylation (OXPHOS) to promote proliferation and maintenance. Cancer cells use multiple adaptive mechanisms in response to a hypo-nutrient environment. However, little is known about how cancer mitochondria are involved in the ability of these cells to adapt to a hypo-nutrient environment. Oncogenic HRas leads to suppression of the mitochondrial oxygen consumption rate (OCR), but oxygen consumption is essential for tumorigenesis. We found that in oncogenic HRas transformed cells, serum depletion reversibly increased the OCR and membrane potential. Serum depletion promoted a cancer stem cell (CSC)-like phenotype, indicated by an increase in CSC markers expression and resistance to anticancer agents. We also found that nitric oxide (NO) synthesis was significantly induced after serum depletion and that NO donors modified the OCR. An NOS inhibitor, SEITU, inhibited the OCR and CSC gene expression. It also reduced anchorage-independent growth by promoting apoptosis. In summary, our data provide new molecular findings that serum depletion induces NO synthesis and promotes mitochondrial OXPHOS, leading to tumor progression and a CSC phenotype. These results suggest that mitochondrial OCR inhibitors can be used as therapy against CSC.
• Plasma Metabolites Predict Severity of Depression and Suicidal Ideation in Psychiatric Patients-A Multicenter Pilot Analysis.
  Daiki Setoyama; Takahiro A Kato; Ryota Hashimoto; Hiroshi Kunugi; Kotaro Hattori; Kohei Hayakawa; Mina Sato-Kasai; Norihiro Shimokawa; Sachie Kaneko; Sumiko Yoshida; Yu-Ichi Goto; Yuka Yasuda; Hidenaga Yamamori; Masahiro Ohgidani; Noriaki Sagata; Daisuke Miura; Dongchon Kang; Shigenobu Kanba
  PloS one, 11, 12, e0165267, 2016年, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Evaluating the severity of depression (SOD), especially suicidal ideation (SI), is crucial in the treatment of not only patients with mood disorders but also psychiatric patients in general. SOD has been assessed on interviews such as the Hamilton Rating Scale for Depression (HAMD)-17, and/or self-administered questionnaires such as the Patient Health Questionnaire (PHQ)-9. However, these evaluation systems have relied on a person's subjective information, which sometimes lead to difficulties in clinical settings. To resolve this limitation, a more objective SOD evaluation system is needed. Herein, we collected clinical data including HAMD-17/PHQ-9 and blood plasma of psychiatric patients from three independent clinical centers. We performed metabolome analysis of blood plasma using liquid chromatography mass spectrometry (LC-MS), and 123 metabolites were detected. Interestingly, five plasma metabolites (3-hydroxybutyrate (3HB), betaine, citrate, creatinine, and gamma-aminobutyric acid (GABA)) are commonly associated with SOD in all three independent cohort sets regardless of the presence or absence of medication and diagnostic difference. In addition, we have shown several metabolites are independently associated with sub-symptoms of depression including SI. We successfully created a classification model to discriminate depressive patients with or without SI by machine learning technique. Finally, we produced a pilot algorithm to predict a grade of SI with citrate and kynurenine. The above metabolites may have strongly been associated with the underlying novel biological pathophysiology of SOD. We should explore the biological impact of these metabolites on depressive symptoms by utilizing a cross species study model with human and rodents. The present multicenter pilot study offers a potential utility for measuring blood metabolites as a novel objective tool for not only assessing SOD but also evaluating therapeutic efficacy in clinical practice. In addition, modification of these metabolites by diet and/or medications may be a novel therapeutic target for depression. To clarify these aspects, clinical trials measuring metabolites before/after interventions should be conducted. Larger cohort studies including non-clinical subjects are also warranted to clarify our pilot findings.
• Disruption of mitochondrial fission in the liver protects mice from diet-induced obesity and metabolic deterioration.
  Lixiang Wang; Takaya Ishihara; Yuta Ibayashi; Keita Tatsushima; Daiki Setoyama; Yuki Hanada; Yukina Takeichi; Shohei Sakamoto; Sadaki Yokota; Katsuyoshi Mihara; Dongchon Kang; Naotada Ishihara; Ryoichi Takayanagi; Masatoshi Nomura
  Diabetologia, 58, 10, 2371, 80, 2015年10月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, AIM/HYPOTHESIS: Mitochondria and the endoplasmic reticulum (ER) physically interact by close structural juxtaposition, via the mitochondria-associated ER membrane. Inter-organelle communication between the ER and mitochondria has been shown to regulate energy metabolism and to be central to the modulation of various key processes such as ER stress. We aimed to clarify the role of mitochondrial fission in this communication. METHODS: We generated mice lacking the mitochondrial fission protein dynamin-related protein 1 (DRP1) in the liver (Drp1LiKO mice). RESULTS: Drp1LiKO mice showed decreased fat mass and were protected from high-fat diet (HFD)-induced obesity. Analysis of liver gene expression profiles demonstrated marked elevation of ER stress markers. In addition, we observed increased expression of the fibroblast growth factor 21 (FGF21) gene through induction of activating transcription factor 4, master regulator of the integrated stress response. CONCLUSIONS/INTERPRETATION: Disruption of mitochondrial fission in the liver provoked ER stress, while inducing the expression of FGF21 to increase energy expenditure and protect against HFD-induced obesity.
• [Mass Spectrometry Based Metabolic Study; Application to Mitochondria Dysfunction Model for Biomarker Discovery].
  Daiki Setoyama; Dongchon Kang
  Fukuoka igaku zasshi = Hukuoka acta medica, 106, 10, 267, 72, 2015年10月, ［査読有り］, ［国内誌］
  日本語, 研究論文（学術雑誌）
• Identification of 3-methylbutanoyl glycosides in green Coffea arabica beans as causative determinants for the quality of coffee flavors.
  Keiko Iwasa; Daiki Setoyama; Hiroaki Shimizu; Harumichi Seta; Yoshinori Fujimura; Daisuke Miura; Hiroyuki Wariishi; Chifumi Nagai; Koichi Nakahara
  Journal of agricultural and food chemistry, 63, 14, 3742, 51, 2015年04月15日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, The quality of coffee green beans is generally evaluated by the sensory cupping test, rather than by chemical compound-based criteria. In this study, we examined the relationship between metabolites and cupping scores for 36 varieties of beans, using a nontargeted LC-MS-based metabolic profiling technique. The cupping score was precisely predicted with the metabolic information measured using LC-MS. Two markers that strongly correlated with high cupping scores were determined to be isomers of 3-methylbutanoyl disaccharides (3MDs; 0.01-0.035 g/kg of beans) by spectroscopic analyses after purification, and one of them was a novel structure. Further, both the 3MDs were determined to be precursors of 3-methylbutanoic acid that enhance the quality of coffee. The applicability of 3MDs as universal quality indicators was validated with another sample set. It was concluded that 3MDs are the causative metabolites determining beverage quality and can be utilized for green bean selection and as key compounds for improving the beverage quality.
• Direct induction of ramified microglia-like cells from human monocytes: dynamic microglial dysfunction in Nasu-Hakola disease.
  Masahiro Ohgidani; Takahiro A Kato; Daiki Setoyama; Noriaki Sagata; Ryota Hashimoto; Kazue Shigenobu; Tetsuhiko Yoshida; Kohei Hayakawa; Norihiro Shimokawa; Daisuke Miura; Hideo Utsumi; Shigenobu Kanba
  Scientific reports, 4, 4957, 4957, 2014年05月14日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Microglia have been implicated in various neurological and psychiatric disorders in rodent and human postmortem studies. However, the dynamic actions of microglia in the living human brain have not been clarified due to a lack of studies dealing with in situ microglia. Herein, we present a novel technique for developing induced microglia-like (iMG) cells from human peripheral blood cells. An optimized cocktail of cytokines, GM-CSF and IL-34, converted human monocytes into iMG cells within 14 days. The iMG cells have microglial characterizations; expressing markers, forming a ramified morphology, and phagocytic activity with various cytokine releases. To confirm clinical utilities, we developed iMG cells from a patient of Nasu-Hakola disease (NHD), which is suggested to be directly caused by microglial dysfunction, and observed that these cells from NHD express delayed but stronger inflammatory responses compared with those from the healthy control. Altogether, the iMG-technique promises to elucidate unresolved aspects of human microglia in various brain disorders.
• Mass spectrometry-based metabolic profiling of gemcitabine-sensitive and gemcitabine-resistant pancreatic cancer cells.
  Yoshinori Fujimura; Naoki Ikenaga; Kenoki Ohuchida; Daiki Setoyama; Miho Irie; Daisuke Miura; Hiroyuki Wariishi; Masaharu Murata; Kazuhiro Mizumoto; Makoto Hashizume; Masao Tanaka
  Pancreas, 43, 2, 311, 8, 2014年03月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, OBJECTIVES: Gemcitabine resistance (GR) is one of the critical issues for therapy for pancreatic cancer, but the mechanism still remains unclear. Our aim was to increase the understanding of GR by metabolic profiling approach. METHODS: To establish GR cells, 2 human pancreatic cancer cell lines, SUIT-2 and CAPAN-1, were exposed to increasing concentration of gemcitabine. Both parental and chemoresistant cells obtained by this treatment were subjected to metabolic profiling based on liquid chromatography-mass spectrometry. RESULTS: Multivariate statistical analyses, both principal component analysis and orthogonal partial least squares discriminant analysis, distinguished metabolic signature of responsiveness and resistance to gemcitabine in both SUIT-2 and CAPAN-1 cells. Among significantly different (P < 0.005) metabolite peaks between parental and GR cells, we identified metabolites related to several metabolic pathways such as amino acid, nucleotide, energy, cofactor, and vitamin pathways. Decreases in glutamine and proline levels as well as increases in aspartate, hydroxyproline, creatine, and creatinine levels were observed in chemoresistant cells from both cell lines. CONCLUSIONS: These results suggest that metabolic profiling can isolate distinct features of pancreatic cancer in the metabolome of gemcitabine-sensitive and GR cells. These findings may contribute to the biomarker discovery and an enhanced understanding of GR in pancreatic cancer.
• Metabolomics reveals that carnitine palmitoyltransferase-1 is a novel target for oxidative inactivation in human cells.
  Daiki Setoyama; Yoshinori Fujimura; Daisuke Miura
  Genes to cells : devoted to molecular & cellular mechanisms, 18, 12, 1107, 19, 2013年12月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Oxidative dysfunction in the metabolism has long been implicated in diverse biological disorders. Although a substantial number of metabolic enzymes are targeted for inactivation by oxidative stress, identifying those targets remains difficult due to a lack of comprehensive observations of the metabolism acting through the stress response. We herein developed a metabolomics strategy using integrative liquid chromatography-mass spectrometry (LC-MS) and observing rapid metabolomic changes in response to hydrogen peroxide (H2 O2 )-induced oxidative stress in HeLa cells. Among the many metabolite changes detected, the most characteristic metabolites uniquely indicated carnitine palmitoyltransferase-1 (CPT1), the critical enzyme for mitochondrial β-oxidation of long-chain fatty acids, to be a target for oxidative inactivation. We showed that the enzymatic activity of CPT1 significantly declined by H2 O2 in several human cells. Interestingly, the inactivation was shown to be a direct effect of H2 O2 in vitro, but substantially occurred when cells were cultured with some reagents that generate reactive oxygen species (ROS). Thus, our results suggest the generality of CPT1 inhibition under various stress conditions associated with ROS generation, providing an insight into a mechanism for oxidative dysfunction in mitochondrial metabolism. Our metabolome data additionally suggest that certain methyltransferase(s) may be targets of oxidative stress as well.
• High-throughput metabolic profiling of diverse green Coffea arabica beans identified tryptophan as a universal discrimination factor for immature beans.
  Daiki Setoyama; Keiko Iwasa; Harumichi Seta; Hiroaki Shimizu; Yoshinori Fujimura; Daisuke Miura; Hiroyuki Wariishi; Chifumi Nagai; Koichi Nakahara
  PloS one, 8, 8, e70098, 2013年, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, The maturity of green coffee beans is the most influential determinant of the quality and flavor of the resultant coffee beverage. However, the chemical compounds that can be used to discriminate the maturity of the beans remain uncharacterized. We herein analyzed four distinct stages of maturity (immature, semi-mature, mature and overripe) of nine different varieties of green Coffea arabica beans hand-harvested from a single experimental field in Hawaii. After developing a high-throughput experimental system for sample preparation and liquid chromatography-mass spectrometry (LC-MS) measurement, we applied metabolic profiling, integrated with chemometric techniques, to explore the relationship between the metabolome and maturity of the sample in a non-biased way. For the multivariate statistical analyses, a partial least square (PLS) regression model was successfully created, which allowed us to accurately predict the maturity of the beans based on the metabolomic information. As a result, tryptophan was identified to be the best contributor to the regression model; the relative MS intensity of tryptophan was higher in immature beans than in those after the semi-mature stages in all arabica varieties investigated, demonstrating a universal discrimination factor for diverse arabica beans. Therefore, typtophan, either alone or together with other metabolites, may be utilized for traders as an assessment standard when purchasing qualified trading green arabica bean products. Furthermore, our results suggest that the tryptophan metabolism may be tightly linked to the development of coffee cherries and/or beans.
• Human MTH3 (NUDT18) protein hydrolyzes oxidized forms of guanosine and deoxyguanosine diphosphates: comparison with MTH1 and MTH2.
  Yasumitsu Takagi; Daiki Setoyama; Riyoko Ito; Hiroyuki Kamiya; Yuriko Yamagata; Mutsuo Sekiguchi
  The Journal of biological chemistry, 287, 25, 21541, 9, 2012年06月15日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Most of the proteins carrying the 23-residue MutT-related sequence are capable of hydrolyzing compounds with a general structure of nucleoside diphosphate linked to another moiety X and are called the Nudix hydrolases. Among the 22 human Nudix proteins (identified by the sequence signature), some remain uncharacterized as enzymes without a defined substrate. Here, we reveal that the NUDT18 protein, whose substrate was unknown, can degrade 8-oxo-7,8-dihydroguanine (8-oxo-Gua)-containing nucleoside diphosphates to the monophosphates. Because this enzyme is closely related to MTH1 (NUDT1) and MTH2 (NUDT15), we propose that it should be named MTH3. Although these three human proteins resemble each other in their sequences, their substrate specificities differ considerably. MTH1 cleaves 8-oxo-dGTP but not 8-oxo-dGDP, whereas MTH2 can degrade both 8-oxo-dGTP and 8-oxo-dGDP, although the intrinsic enzyme activity of MTH2 is considerably lower than that of MTH1. On the other hand, MTH3 is specifically active against 8-oxo-dGDP and hardly cleaves 8-oxo-dGTP. Other types of oxidized nucleoside diphosphates, 2-hydroxy-dADP and 8-hydroxy-dADP, were also hydrolyzed by MTH3. Another notable feature of the MTH3 enzyme is its action toward the ribonucleotide counterpart. MTH3 can degrade 8-oxo-GDP as efficiently as 8-oxo-dGDP, which is in contrast to the finding that MTH1 and MTH2 show a limited activity against the ribonucleotide counterpart, 8-oxo-GTP. These three enzymes may function together to help maintain the high fidelity of DNA replication and transcription under oxidative stress.
• Cleavage of oxidized guanine nucleotide and ADP sugar by human NUDT5 protein.
  Riyoko Ito; Mutsuo Sekiguchi; Daiki Setoyama; Yoshimichi Nakatsu; Yuriko Yamagata; Hiroshi Hayakawa
  Journal of biochemistry, 149, 6, 731, 8, 2011年06月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, MutT-related proteins, including Escherichia coli MutT and the human MTH1 (NUDT1), degrade 8-oxo-7, 8-dihydrodeoxyguanosine triphosphate (8-oxo-dGTP) to 8-oxo-dGMP and thereby prevent mutations caused by the misincorporation of 8-oxoguanine into DNA. The human NUDT5, which has an intrinsic activity to cleave ADP sugars to AMP and sugar phosphate, possesses the ability to degrade 8-oxo-dGDP to the monophosphate. Since 8-oxo-dGDP and 8-oxo-dGTP are interconvertible by cellular enzymes, NUDT5 has the potential to prevent errors during DNA replication. The two activities associated with NUDT5 exhibit different pH dependencies; the optimum for the cleavage of ADP ribose is pH 7-9, while that for 8-oxo-dGDPase is around pH 10. The kinetic parameters for the two types of reactions indicated that ADP ribose is a better substrate for NUDT5 compared with oxidized guanine nucleotides. The 8-oxo-dGDP cleavage was competitively inhibited by ADP ribose and its reaction product, AMP, and in reverse, the cleavage of ADP ribose was inhibited by 8-oxo-dGDP. These results imply that the two types of substrates may share the same binding site for catalysis.
• Molecular actions of Escherichia coli MutT for control of spontaneous mutagenesis.
  Daiki Setoyama; Riyoko Ito; Yasumitsu Takagi; Mutsuo Sekiguchi
  Mutation research, 707, 1-2, 9, 14, 2011年02月10日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, MutT protein of Escherichia coli hydrolyzes oxidized guanine nucleotides, 8-oxo-dGTP and 8-oxoGTP, to the corresponding monophosphates, thereby preventing misincorporation of 8-oxoguanine into DNA and RNA, respectively. Although the biological significance of the MutT has been established, how MutT protein actually works in vivo remains to be elucidated. The current study shows the molecular behavior of the MutT protein in vivo and in vitro with special reference to control of spontaneous mutagenesis. A single E. coli cell carries about 70-75 molecules of the MutT protein and that this number does not change even when the cells were cultured in anaerobic and hyper-oxidative conditions. Conditional gene silencing analyses revealed that about a half number of MutT molecules are needed for keeping the spontaneous mutation frequency at the normal level. The MutT functions are not needed under anaerobic condition, yet the level of the MutT protein in cell is kept constant, probably for preparing for sudden changes of oxygen pressure. There is a possibility that MutT functions in close association with other proteins, and evidence is presented that MutT protein can interact with some proteins in vivo.
• Mechanism of degradation of CPEB during Xenopus oocyte maturation.
  Daiki Setoyama; Masakane Yamashita; Noriyuki Sagata
  Proceedings of the National Academy of Sciences of the United States of America, 104, 46, 18001, 6, 2007年11月13日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, CPEB, a cytoplasmic polyadenylation element-binding protein, plays an important role in translational control of maternal mRNAs in early animal development. During Xenopus oocyte maturation, CPEB undergoes a Cdc2-mediated phosphorylation- and ubiquitin-dependent degradation that is required for proper entry into meiosis II. However, the precise mechanism of CPEB degradation, including the identity of the responsible E3 ubiquitin ligase, is not known. Here, we show that the SCF(beta-TrCP) E3 ubiquitin ligase complex targets CPEB for degradation during Xenopus oocyte maturation. beta-TrCP, the F-box protein of SCF(beta-TrCP), specifically binds to a sequence (190)TSGFSS(195) (termed here the TSG motif) of CPEB, thereby targeting CPEB for degradation. beta-TrCP binding depends on phosphorylation of Thr-190, Ser-191, and Ser-195 in the TSG motif. Among these residues, Ser-191 is phosphorylated by the Polo-like kinase Plx1, which binds CPEB at a specific Thr-125 residue prephosphorylated by Cdc2. Finally, Cdc2-mediated phosphorylation of other multiple Ser residues, previously implicated in CPEB degradation, is required for both Thr-125 phosphorylation and beta-TrCP binding, presumably causing conformational changes of CPEB. We propose that Cdc2 and Plx1 sequentially phosphorylate CPEB and target it for SCF(beta-TrCP)-dependent degradation in Xenopus oocytes. We suggest that many other proteins carrying the TSG-like motif may be targeted by SCF(beta-TrCP).

• 特許第7819901号 名称 ひきこもりのバイオマーカー、ひきこもり識別支援方法
  瀬戸山 大樹; 加藤 隆弘; 康 東天; 松島 敏夫, 2026年02月
• Blood-Based Metabolomic Biomarkers and Personality Traits in Major Depressive Disorder: Toward an Integrated Framework for Precision Psychiatry.
  Setoyama D.; Matsushima T.; Kato TA, Advances in Experimental Medicine and Biology., 2026年
• メンタルヘルスの社会的課題に応える計測技術：産学連携による社会実装への道筋
  瀬戸山 大樹; 斎木 秀和; 早川 禎宏; 加藤 隆弘, 島津評論, 2026年
• 部内コードと標準コードのマッピング〜生成AI導入による効率化の可能性〜
  瀬戸山 大樹, 令和6年度AMED中島班「医療施設における標準コードの効率的なマッピング手法に関する調査および実証研究」, 2025年03月
• うつ病の客観的血液指標を求めて-メタボローム解析
  康 東天; 瀬戸山 大樹; 加藤 隆弘, 医学のあゆみ 医歯薬出版, 292, 13, 2025年
• 2. PCT/JP2025/30941 名称 リスク値の検出方法、ＩＣＡＮＳ発症可能性の予測を補助する方法、医薬およびＩＣＡＮＳ発症予測用キット
  瀬戸山 大樹; 野見山 倫子; 國崎 祐哉, 2025年
• Toward objective diagnostics in depression: Blood metabolomic analysis focusing on depressive states, suicidal ideation, and personality traits.
  Setoyama D.; Kato TA; Kunisaki Y.; Kang D., LabMed Discovery., 2025年
• うつ病の血液バイオマーカーを探索する- メタボロミクス
  松島 敏夫; 瀬戸山 大樹; 加藤 隆弘, 医学のあゆみ 医歯薬出版, 292, 13, 2025年
• ひきこもりの生物学的理解に向けたバイオマーカー探索の現状と今後の指針
  瀬戸山 大樹, 医学のあゆみ 医歯薬出版, 293, 2, 2025年
• CAR-T療法における脳脊髄液を用いたICANS発症のバイオマーカー探索と今後の展望 検査と技術
  野見山 倫子; 瀬戸山 大樹; 國崎 祐哉, 2025年
• 特願2023-117877 名称 精神的ストレスの補助診断方法
  瀬戸山 大樹、加藤 隆弘, 2023年
• 慢性心不全モデルマウスの病態解析と治療戦略
  平井 遥; 八木 美佳子; 都 由羅; 瀬戸山 大樹; 堀田 多恵子; 内海 健, 臨床化学, 52, 1, 47, 50, 2023年
• 精神疾患の血液メタボローム解析研究 うつ病から社会的ひきこもりまで
  加藤 隆弘; 松島 敏夫; 瀬戸山 大樹, 日本生物学的精神医学会誌, 34, 1, 47, 50, 2023年
• 経口糖尿病薬SGLT-2阻害薬服用中の高齢女性患者で気尿を呈した尿内発酵症候群の1例
  土居 壽孝; 瀬戸山 大樹; 宮本 哲哉; 康 東天, 臨床と研究, 99, 4, 97, 100, 2022年
• ミトコンドリア代謝を知るためのメタボロミクス
  瀬戸山 大樹, 生化学 日本生化学会, 94, 2, 159, 169, 2022年
• 「メンタルヘルスの臨床検査」に向けた血液バイオマーカー探索
  瀬戸山 大樹, 臨床化学 日本臨床化学会, 51, 3, 161, 168, 2022年
• 気分障害をターゲットとしたメタボローム解析研究-九州大学病院でのトライアル
  松島 敏夫; 瀬戸山 大樹; 加藤 隆弘, 臨床精神医学 国際医書出版, 51, 1121, 1129, 2022年
• 特願2021-66338 名称 分析方法および前処理装置
  瀬戸山 大樹; 加藤 隆弘; 康 東天; 神庭 重信, 2021年
• 特願2020-44908 コエンザイムQ産生促進剤及びコエンザイムQ産生促進方法
  瀬戸山 大樹, 2020年
• 臨床検査診断支援(DSS)システムの開発を目指した検査値データの活用事例について
  瀬戸山 大樹, 医療検査と自動化, 45, suppl.2, 165, 171, 2020年
• 診断バイオマーカー-うつ病の血液バイオマーカー開発の試み-
  瀬戸山 大樹; 加藤 隆弘, 精神科（心療内科）科学評論社, 37, 6, 592, 598, 2020年
• 質量分析による代謝解析アプリケーション〜ミトコンドリア障害を捉えるバイオマーカー探索〜
  瀬戸山 大樹, 福岡医学雑誌, 106, 10, 267, 272, 2015年
• 特許第6882776号 名称 うつ病診断用バイオマーカー及びその使用
  瀬戸山 大樹; 加藤 隆弘; 康 東天; 神庭 重信, 2000年

• Schizoid Personality, Hikikomori, and Blood Biomarkers: A Focus on Lipidomics and Metabolomics.
  Hayakawa K.; Setoyama D.; Kato TA
  Handbook of the Biology and Pathology of Mental Disorders. Springer International Publishing Cham; 2024. p.1–17., 2024年
• 実験医学別冊 決定版質量分析活用スタンダード〜代謝物からタンパク質 「LC-MS/MS ターゲット分析 臨床バイオマーカー探索を始める前に」
  野見山 倫子; 瀬戸山 大樹
  羊土社, 2023年
• 精神医学領域の論文を読みこなすキーワード100! 「オミクス解析」
  瀬戸山 大樹; 加藤 隆弘
  新興医学出版社, 2022年

• Exploring Blood-Based Biomarkers for Hikikomori in Japan. UK-Japan Neuroscience Symposium
  SETOYAMA Daiki
  Cambridge (poster presentation), 2026年03月
• ひきこもりの状態評価および症状改善予測を可能にする血液バイオマーカー探索
  瀬戸山 大樹
  第９９回日本薬理学会年会 企画シンポジウム 仙台, 2026年03月
• 心の健康を可視化する〜血液✕AIで挑むメンタルヘルス健診の社会実装〜
  瀬戸山 大樹
  九州大学学術研究都市セミナーin東京, 2025年12月
• 気分障害・ひきこもりの血液バイオマーカー：トリプトファン代謝物とマウス・ヒト横断研究からの知見
  第６５回日本臨床化学会年次学術集会 名古屋 第４３回日本トリプトファン研究会学術集会, 2025年11月
• LM Studioで学ぶlocal LLMによる検査データ解析入門：ナラティブ生成と自然言語解析体験
  瀬戸山 大樹
  日本医療検査科学会第57回大会、横浜, 2025年10月
• B細胞腫瘍に対するCAR-T治療の副作用（神経毒性）を予測する髄液中タンパク質マーカーの同定と実用化
  瀬戸山 大樹
  新技術説明会 Web, 2025年08月
• ひきこもり支援における臨床検査の新展開：症状改善に伴う血液成分モニタリング法の開発
  瀬戸山 大樹
  第１２１回日本精神神経学会学術総会 神戸, 2025年06月
• 国試準備期間の学生の心理ストレスとメンタルヘルスの血液バイオマーカーの統合解析：個別化ストレスマネジメントへの応用
  瀬戸山 大樹; 福應温; 國崎祐哉; 加藤隆弘
  日本薬学会第１４５回年会 福岡, 2025年03月
• 部内コードと標準コードのマッピングの課題〜生成AIを用いたマッピング事例〜
  瀬戸山 大樹
  第５６回日本医療検査科学会年会 横浜, 2024年10月
• 問診と血液バイオマーカーを用いた客観的ストレス診断支援ビジネス
  瀬戸山 大樹; 加藤隆弘
  純真学園大学・学園祭・講演会 福岡, 2024年10月
• 質量分析を備えた大学病院検査部の使命〜検査実装とバイオマーカー探索〜」
  瀬戸山 大樹
  第４９回日本医用マススペクトル学会年会、京都, 2024年09月
• 質量分析によるメンタルヘルスの血液検査の社会実装に向けて
  瀬戸山 大樹
  第６４回日本臨床化学学会年次学術集会 宇都宮, 2024年08月
• Establishing Objective Metrics Utilizing Blood Markers from Hikikomori Individuals
  Daiki Setoyama (Invited speaker)
  A Study in Progress UK-Japan Hikikomori Workshop 2024, Fukuoka, 2024年06月
• ひきこもりの客観的血液指標の探索
  瀬戸山 大樹
  第４５回日本生物学的精神医学会年会、沖縄, 2023年11月
• 医療ビッグデータの研究事例を生成系AIで調べてみたら
  瀬戸山 大樹
  第５５回日本医療検査科学会年会 横浜, 2023年10月
• Deciphering modes of action of longevity-associated metabolites on mitochondria by multi-omics approach
  Daiki Setoyama (Invited speaker)
  CNU/KAIST seminar, Korea Daejon, 2023年09月
• 質量分析プラットフォームによる臨床オミックス研究
  瀬戸山 大樹
  第３回眼科オミックス研究会 web, 2023年09月
• Development of blood biomarkers of hikikomori
  Daiki Setoyama(Invited speaker
  Italy-Japan Hikikomori Research Workshop 2023, Fukuoka, 2023年04月
• 有機酸代謝異常症の自動化分析法の確立とメンタルヘルスの臨床検査への挑戦
  瀬戸山 大樹
  第61回日本臨床化学会年次学術集会, 福岡, 2021年11月
• Compartmentalized Metabolomics Reveals a Substantial Role of NAD+ in Mitochondrial DNA Maintenance
  Daiki Setoyama(Invited speaker
  The 11th International Congress of Diabetes and Metabolism and The 13th AASD Scientific Meeting, Web, 2021年10月
• 最近の研究事例から学ぶ、臨床検査の機械学習法〜COVID-19診断・重症化/死亡率予測アルゴリズム作成に関する文献を題材に〜
  瀬戸山 大樹
  日本臨床検査自動化学会第53回大会, 2021年10月
• 臨床検査データ解析の実際-Rを用いた解析の基本・応用・発展
  瀬戸山 大樹
  日本臨床検査自動化学会第52回大会, 2020年10月
• オルガネラメタボロミクスと臨床メタボロミクス
  瀬戸山 大樹
  島津製作所-名古屋大学 Zoomセミナー, 2020年06月
• 臨床検査データ解析の実際
  瀬戸山 大樹
  日本臨床検査自動化学会第51回大会, 横浜, 2019年10月
• 臨床検査のデータサイエンス職能拡大推進事業「診療情報、医療情報の知識・技術によるデータの活用・医療への貢献」,
  瀬戸山 大樹
  福岡, 2019年09月
• LCMS-based Plasma Metabolite Biomarker Analysis of Major Depressive Disorders: A Novel Approach.
  Daiki Setoyama(Invited speaker)
  Shimadzu Global Innovation Summit, Kyoto, 2019年07月
• 臨床検査のデータサイエンス〜現状と課題〜 観察研究のススメ〜
  瀬戸山 大樹
  第９回シンポジウム福岡, 2019年06月
• Rで広がるデータ解析の地平
  瀬戸山 大樹
  福岡県保健環境研究所集談会, 2019年02月
• 臨床検査値の機械学習応用 病院データベース利用に関するプラクティカルな課題
  瀬戸山 大樹; 堀田多恵子; 康東天
  日本臨床検査自動化学会第５０回大会 神戸, 2018年10月
• Mass Spectrometry-Based Omics Approach Reveals Multifaceted Roles of Mitochondrial Metabolism in the Drug Target and Maintaining its Own DNA
  Daiki Setoyama(Invited speaker)
  第二回日本循環器学会基礎研究フォーラム, 奈良, 2018年09月
• うつ病バイオマーカー探索のための新たな血しょう代謝物の層別化解析法
  瀬戸山 大樹
  第５８回日本臨床化学年次学術集会 名古屋, 2018年08月
• メンタルヘルスの臨床検査における質量分析の役割
  瀬戸山 大樹
  第５８回日本臨床化学会年次学術集会 名古屋, 2018年08月
• 病院検査部における医療情報リテラシー教育〜機械学習・AI技術による臨床検査診断支援システムの実現を目指して
  瀬戸山 大樹
  平成29年度 九州大学 数理・データサイエンスに関する教育・研究支援プログラム 福岡, 2018年03月

• 日本プロテオーム学会会員
• 日本細胞外小胞学会会員
• 日本臨床化学会会員
• 日本分子生物学会会員
• 日本医療検査科学会 医療情報委員会委員

• 双極症うつ病相におけるケタミン治療の有効性検証と、効果に関連する代謝物質の探索
  科学研究費助成事業
  2025年04月01日 - 2028年03月31日
  櫻井 準; 瀬戸山 大樹; 立森 久照; 加藤 隆弘
  日本学術振興会, 基盤研究(C), 杏林大学, 25K10822