Research activity information

• Neutrophil gene expression in COVID-19 patients with acute respiratory distress syndrome

  Hiroshi Ito, Masakazu Ishikawa, Jumpei Yoshimura, Yuchen Liu, Shuhei Sakakibara, Fuminori Sugihara, Hisatake Matsumoto, Haruhiko Hirata, Hiroshi Ogura, Jun Oda, Daisuke Okuzaki

  Background Although an increase in neutrophil count has been observed in patients with coronavirus disease 2019 (COVID-19), the relationship between the systemic neutrophil transcriptome and clinical course of COVID-19 remains unclear. Hence, we examined the relationship between the clinical course and RNA sequencing analysis results in COVID-19 patients. Methods Peripheral blood samples were obtained from 28 patients with COVID-19-associated ARDS and 16 healthy controls. Bulk RNA sequencing was performed, and clustering analysis was used to explore relationships between gene expression and clinical characteristics. In a separate cohort, neutrophils were isolated from the peripheral blood of five COVID-19 patients with ARDS for single-cell RNA sequencing to further characterize the neutrophil subpopulations. Results In bulk RNA sequencing analysis, COVID-19 patients with ARDS had elevated gene expression associated with neutrophils compared with healthy controls.Clustering analysis revealed no differences in the clinical characteristics of COVID-19 patients with ARDS. In the single-cell RNA sequencing analysis, clustering analysis showed that the patients were divided into two groups: those who could be weaned from the ventilator within 28 days and those who could not be weaned. Conclusion These findings indicate that differences in neutrophil gene expression may have important clinical implications. This study may support the exploratory identification of genomic factors, such as neutrophil gene expression, that are relevant to clinical parameters.

  Frontiers Media SA, Nov. 2025, Frontiers in Immunology, 16

  Scientific journal


• Loss of PI3Kδ Activity Drives Autoimmune Colitis by Impairing Extrathymic Treg Differentiation.

  Ee Lyn Lim, Yamin Qian, Fuminori Sugihara, Atsushi Tanaka, Shimon Sakaguchi

  Peripherally derived regulatory T cells (pTregs) have a prominent role in maintaining intestinal immune homeostasis. In cases of phosphoinositide-3-kinase δ (PI3Kδ) inactivation, such as in patients receiving PI3Kδ inhibitor idelalisib as a cancer treatment, breakdown of intestinal immune tolerance occurs frequently in the form of diarrhea and colon inflammation. In a mouse model of systemic PI3Kδ inactivation, both enhancement of antitumor immunity and colitis have been described as a result of Treg impairment. However, in view of the critical role for Tregs in the prevention of systemic autoimmunity, the basis for such tissue-restricted breach of immune tolerance upon loss of PI3Kδ function is not yet understood. We report here that mice lacking PI3Kδ activity do not suffer a general defect in Treg immunosuppression, but specifically fail to develop Helios- pTregs in the colon. We demonstrate reduced extrathymic Treg induction, in vitro and in vivo, from naïve CD4+ T cells with inactive PI3Kδ, along with dysregulation of a tissue-resident phenotype. These results suggest a nonredundant role for PI3Kδ-dependent pTreg differentiation in maintaining tolerance to commensal microbial antigens in the gut.

  Oct. 2025, European journal of immunology, 55(10) (10), e70069, English, International magazine

  Scientific journal


• Longitudinal proteomic analysis of pathophysiology in plasma and bronchoalveolar lavage fluid of patients with ARDS.

  Yumi Mitsuyama, Hisatake Matsumoto, Fuminori Sugihara, Satoshi Fujimi, Hiroshi Ogura, Jun Oda

  BACKGROUND: Acute respiratory distress syndrome (ARDS) remains a significant clinical challenge, and its pathogenesis is not fully understood. Proteomic analyses of plasma and bronchoalveolar lavage fluid (BALF) in patients with ARDS have been performed to uncover diagnostic and prognostic markers, although previous studies have not adequately focused on longitudinal comparison of biomarkers. This study aimed to elucidate the proteomic profiles of patients with ARDS in the acute and subacute phases to better understand the pathophysiological progression of ARDS. METHODS: This was a single-center, prospective, observational study of adult patients with ARDS in whom plasma and BALF samples were collected in the acute and subacute phases of ARDS and comprehensive proteins were identified and analyzed by mass spectrometry. RESULTS: Plasma and BALF were collected from 21 ARDS patients and plasma from 24 healthy donors, from which 694 plasma proteins and 2017 BALF proteins were analyzed. Processes related to coagulation and complement commonly activated in plasma and BALF were more pronounced in the acute phase than in the subacute phase. In BALF in the acute phase, pathways related to humoral and immune responses were activated, whereas processes related to chaperones and protein folding were suppressed. IPA analysis showed that B cell receptor signaling was most activated, whereas heat shock protein 90 (HSP90) chaperone cycle, protein folding, and other pathways associated with cellular stress responses and proper protein processing were suppressed. The most activated upstream regulator was interferon gamma (IFN-γ) and the most suppressed was notch receptor 1 (NOTCH1). CONCLUSIONS: The proteomics of plasma and BALF from patients with ARDS were compared in both the acute and subacute phases. In BALF in the acute phase, humoral immunity, mainly B-cell receptor signaling, was activated, whereas the HSP90 cycle and protein folding mechanisms were inactivated.

  May 2025, Journal of intensive care, 13(1) (1), 26 - 26, English, International magazine

  Scientific journal


• Retraction for Hiramatsu et al., "Melanin Produced by Bordetella parapertussis Confers a Survival Advantage to the Bacterium during Host Infection".

  Yukihiro Hiramatsu, Takashi Nishida, Dendi Krisna Nugraha, Fuminori Sugihara, Yasuhiko Horiguchi

  Feb. 2025, mSphere, e0107224, English, International magazine

  Scientific journal


• Clonal landscape of autoantibody-secreting plasmablasts in COVID-19 patients.

  Shuhei Sakakibara, Yu-Chen Liu, Masakazu Ishikawa, Ryuya Edahiro, Yuya Shirai, Soichiro Haruna, Marwa Ali El Hussien, Zichang Xu, Songling Li, Yuta Yamaguchi, Teruaki Murakami, Takayoshi Morita, Yasuhiro Kato, Haruhiko Hirata, Yoshito Takeda, Fuminori Sugihara, Yoko Naito, Daisuke Motooka, Chao-Yuan Tsai, Chikako Ono, Yoshiharu Matsuura, James B Wing, Hisatake Matsumoto, Hiroshi Ogura, Masato Okada, Atsushi Kumanogoh, Yukinari Okada, Daron M Standley, Hitoshi Kikutani, Daisuke Okuzaki

  Whereas severe COVID-19 is often associated with elevated autoantibody titers, the underlying mechanism behind their generation has remained unclear. Here we report clonal composition and diversity of autoantibodies in humoral response to SARS-CoV-2. Immunoglobulin repertoire analysis and characterization of plasmablast-derived monoclonal antibodies uncovered clonal expansion of plasmablasts producing cardiolipin (CL)-reactive autoantibodies. Half of the expanded CL-reactive clones exhibited strong binding to SARS-CoV-2 antigens. One such clone, CoV1804, was reactive to both CL and viral nucleocapsid (N), and further showed anti-nucleolar activity in human cells. Notably, antibodies sharing genetic features with CoV1804 were identified in COVID-19 patient-derived immunoglobulins, thereby constituting a novel public antibody. These public autoantibodies had numerous mutations that unambiguously enhanced anti-N reactivity, when causing fluctuations in anti-CL reactivity along with the acquisition of additional self-reactivities, such as anti-nucleolar activity, in the progeny. Thus, potentially CL-reactive precursors may have developed multiple self-reactivities through clonal selection, expansion, and somatic hypermutation driven by viral antigens. Our results revealed the nature of autoantibody production during COVID-19 and provided novel insights into the origin of virus-induced autoantibodies.

  Dec. 2024, Life science alliance, 7(12) (12), English, International magazine

  Scientific journal


• Semaphorin 6D tunes amygdalar circuits for emotional, metabolic, and inflammatory outputs.

  Yoshimitsu Nakanishi, Mayuko Izumi, Hiroaki Matsushita, Yoshihisa Koyama, Diego Diez, Hyota Takamatsu, Shohei Koyama, Masayuki Nishide, Maiko Naito, Yumiko Mizuno, Yuta Yamaguchi, Tomoki Mae, Yu Noda, Kamon Nakaya, Satoshi Nojima, Fuminori Sugihara, Daisuke Okuzaki, Masahito Ikawa, Shoichi Shimada, Sujin Kang, Atsushi Kumanogoh

  Regulated neural-metabolic-inflammatory responses are essential for maintaining physiological homeostasis. However, the molecular machinery that coordinates neural, metabolic, and inflammatory responses is largely unknown. Here, we show that semaphorin 6D (SEMA6D) coordinates anxiogenic, metabolic, and inflammatory outputs from the amygdala by maintaining synaptic homeostasis. Using genome-wide approaches, we identify SEMA6D as a pleiotropic gene for both psychiatric and metabolic traits in human. Sema6d deficiency increases anxiety in mice. When fed a high-fat diet, Sema6d-/- mice display attenuated obesity and enhanced myelopoiesis compared with control mice due to higher sympathetic activity via the β3-adrenergic receptor. Genetic manipulation and spatial and single-nucleus transcriptomics reveal that SEMA6D in amygdalar interneurons is responsible for regulating anxiogenic and autonomic responses. Mechanistically, SEMA6D is required for synaptic maturation and γ-aminobutyric acid transmission. These results demonstrate that SEMA6D is important for the normal functioning of the neural circuits in the amygdala, coupling emotional, metabolic, and inflammatory responses.

  Jul. 2024, Neuron, English, International magazine

  Scientific journal


• Non-classical CD45RBlo memory B-cells are the majority of circulating antigen-specific B-cells following mRNA vaccination and COVID-19 infection.

  David G. Priest, Takeshi Ebihara, Janyerkye Tulyeu, Jonas Søndergaard, Shuhei Sakakibara, Fuminori Sugihara, Shunichiro Nakao, Yuki Togami, Jumpei Yoshimura, Hiroshi Ito, Shinya Onishi, Arisa Muratsu, Yumi Mitsuyama, Hiroshi Ogura, Jun Oda, Daisuke Okuzaki, Hisatake Matsumoto, James B. Wing

  Abstract Resting memory B-cells can be divided into classical and non-classical groups based on differential expression of markers such as CD27 and CD11c, while activated memory B-cells express a combination of markers, making their ontogeny hard to determine. Here by longitudinal analysis of COVID-19, bacterial sepsis, and BNT162b2 mRNA vaccine recipients by mass cytometry and CITE-seq we describe a three-branch structure of resting B-cell memory consisting of “classical” CD45RB⁺ memory and two branches of CD45RB^lo memory further defined by expression of CD23 and CD11c respectively. Stable differences in CD45RB upon activation allowed tracking of activated B-cells and plasmablasts derived from CD45RB⁺ classical and CD45RB^lo non-classical memory B-cells. In both COVID-19 patients and mRNA vaccination, CD45RB^lo B-cells formed the majority of SARS-CoV2 specific memory B-cells and correlated with serum antibodies while CD45RB⁺ memory was most strongly activated by bacterial Sepsis. These results suggest that diverse non-classical CD45RB^lo memory B-cells consisting of branches of CD11c⁺Tbet⁺ and CD23⁺ fractions form a critical part of responses to viral infection and vaccination.

  Research Square Platform LLC, Jan. 2024


• Full-Length Nanopore Sequencing of Circular Rna Landscape in Peripheral Blood Cells Following Sequential Bnt162b2 Mrna Vaccination

  Yu-Chen Liu, Masakazu Ishikawa, Shuhei Sakakibara, Mohamad Al Kadi, D. Motooka, Yoko Naito, Shingo Ito, Yuko Imamura, Hisatake Matsumoto, Fuminori Sugihara, Haruhiko Hirata, Hiroshi Ogura, D. Okuzaki

  Elsevier BV, 2024


• Activation of ILC2s through constitutive IFNγ signaling reduction leads to spontaneous pulmonary fibrosis.

  Natsuko Otaki, Yasutaka Motomura, Tommy Terooatea, S Thomas Kelly, Miho Mochizuki, Natsuki Takeno, Shigeo Koyasu, Miu Tamamitsu, Fuminori Sugihara, Junichi Kikuta, Hideya Kitamura, Yoshiki Shiraishi, Jun Miyanohara, Yuji Nagano, Yuji Saita, Takashi Ogura, Koichiro Asano, Aki Minoda, Kazuyo Moro

  Pulmonary fibrosis (PF), a condition characterized by inflammation and collagen deposition in the alveolar interstitium, causes dyspnea and fatal outcomes. Although the bleomycin-induced PF mouse model has improved our understanding of exogenous factor-induced fibrosis, the mechanism governing endogenous factor-induced fibrosis remains unknown. Here, we find that Ifngr1-/-Rag2-/- mice, which lack the critical suppression factor for group 2 innate lymphoid cells (ILC2), develop PF spontaneously. The onset phase of fibrosis includes ILC2 subpopulations with a high Il1rl1 (IL-33 receptor) expression, and fibrosis does not develop in ILC-deficient or IL-33-deficient mice. Although ILC2s are normally localized near bronchioles and blood vessels, ILC2s are increased in fibrotic areas along with IL-33 positive fibroblasts during fibrosis. Co-culture analysis shows that activated-ILC2s directly induce collagen production from fibroblasts. Furthermore, increased IL1RL1 and decreased IFNGR1 expressions are confirmed in ILC2s from individuals with idiopathic PF, highlighting the applicability of Ifngr1-/-Rag2-/- mice as a mouse model for fibrosis research.

  Dec. 2023, Nature communications, 14(1) (1), 8120 - 8120, English, International magazine

  Scientific journal


• Enhancement of SARS-CoV-2 Infection via Crosslinking of Adjacent Spike Proteins by N-Terminal Domain-Targeting Antibodies

  Tina Lusiany, Tohru Terada, Jun-ichi Kishikawa, Mika Hirose, David Virya Chen, Fuminori Sugihara, Hendra Saputra Ismanto, Floris J. van Eerden, Songling Li, Takayuki Kato, Hisashi Arase, Matsuura Yoshiharu, Masato Okada, Daron M. Standley

  The entry of SARS-CoV-2 into host cells is mediated by the interaction between the spike receptor-binding domain (RBD) and host angiotensin-converting enzyme 2 (ACE2). Certain human antibodies, which target the spike N-terminal domain (NTD) at a distant epitope from the host cell binding surface, have been found to augment ACE2 binding and enhance SARS-CoV-2 infection. Notably, these antibodies exert their effect independently of the antibody fragment crystallizable (Fc) region, distinguishing their mode of action from previously described antibody-dependent infection-enhancing (ADE) mechanisms. Building upon previous hypotheses and experimental evidence, we propose that these NTD-targeting infection-enhancing antibodies (NIEAs) achieve their effect through the crosslinking of neighboring spike proteins. In this study, we present refined structural models of NIEA fragment antigen-binding region (Fab)–NTD complexes, supported by molecular dynamics simulations and hydrogen–deuterium exchange mass spectrometry (HDX-MS). Furthermore, we provide direct evidence confirming the crosslinking of spike NTDs by NIEAs. Collectively, our findings advance our understanding of the molecular mechanisms underlying NIEAs and their impact on SARS-CoV-2 infection.

  MDPI AG, Dec. 2023, Viruses, 15(12) (12), 2421 - 2421

  Scientific journal


• 広範囲熱傷患者におけるプロテオミクス解析 GDF-15と関連パスウェイ

  大西 伸也, 蛯原 健, 松本 寿健, 杉原 文徳, 大須賀 章倫, 小倉 裕司, 織田 順

  (一社)日本救急医学会, Dec. 2023, 日本救急医学会雑誌, 34(12) (12), 752 - 752, Japanese


• Structural basis of Irgb6 inactivation by Toxoplasma gondii through the phosphorylation of switch I

  Hiromichi Okuma, Yumiko Saijo‐Hamano, Hiroshi Yamada, Aalaa Alrahman Sherif, Emi Hashizaki, Naoki Sakai, Takaaki Kato, Tsuyoshi Imasaki, Satoshi Kikkawa, Eriko Nitta, Miwa Sasai, Tadashi Abe, Fuminori Sugihara, Yoshimasa Maniwa, Hidetaka Kosako, Kohji Takei, Daron M. Standley, Masahiro Yamamoto, Ryo Nitta

  Abstract Irgb6 is a priming immune‐related GTPase (IRG) that counteracts Toxoplasma gondii. It is known to be recruited to the low virulent type II T. gondii parasitophorous vacuole (PV), initiating cell‐autonomous immunity. However, the molecular mechanism by which immunity‐related GTPases become inactivated after the parasite infection remains obscure. Here, we found that Thr95 of Irgb6 is prominently phosphorylated in response to low virulent type II T. gondii infection. We observed that a phosphomimetic T95D mutation in Irgb6 impaired its localization to the PV and exhibited reduced GTPase activity in vitro. Structural analysis unveiled an atypical conformation of nucleotide‐free Irgb6‐T95D, resulting from a conformational change in the G‐domain that allosterically modified the PV membrane‐binding interface. In silico docking corroborated the disruption of the physiological membrane binding site. These findings provide novel insights into a T. gondii‐induced allosteric inactivation mechanism of Irgb6.

  Wiley, Nov. 2023, Genes to Cells

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Classification of patients with COVID-19 by blood RNA endotype: a prospective cohort study.

  Jumpei Yoshimura, Yuki Togami, Takeshi Ebihara, Hisatake Matsumoto, Yumi Mitsuyama, Fuminori Sugihara, Haruhiko Hirata, Daisuke Okuzaki, Hiroshi Ogura

  In this study, whole-blood RNAs (prolactin and toll-like receptor 3) involved in the prognosis of patients with COVID-19 were identified. The RNA endotypes classified by these important RNAs highlight the possibility of stratifying the COVID-19 patient population and the need for targeted therapy based on these phenotypes.

  Nov. 2023, Microbiology spectrum, e0264523, English, International magazine

  Scientific journal


• Complement factor D targeting protects endotheliopathy in organoid and monkey models of COVID-19.

  Eri Kawakami, Norikazu Saiki, Yosuke Yoneyama, Chiharu Moriya, Mari Maezawa, Shuntaro Kawamura, Akiko Kinebuchi, Tamaki Kono, Masaaki Funata, Ayaka Sakoda, Shigeru Kondo, Takeshi Ebihara, Hisatake Matsumoto, Yuki Togami, Hiroshi Ogura, Fuminori Sugihara, Daisuke Okuzaki, Takashi Kojima, Sayaka Deguchi, Sebastien Vallee, Susan McQuade, Rizwana Islam, Madhusudan Natarajan, Hirohito Ishigaki, Misako Nakayama, Cong Thanh Nguyen, Yoshinori Kitagawa, Yunheng Wu, Kensaku Mori, Takayuki Hishiki, Tomohiko Takasaki, Yasushi Itoh, Kazuo Takayama, Yasunori Nio, Takanori Takebe

  COVID-19 is linked to endotheliopathy and coagulopathy, which can result in multi-organ failure. The mechanisms causing endothelial damage due to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) remain elusive. Here, we developed an infection-competent human vascular organoid from pluripotent stem cells for modeling endotheliopathy. Longitudinal serum proteome analysis identified aberrant complement signature in critically ill patients driven by the amplification cycle regulated by complement factor B and D (CFD). This deviant complement pattern initiates endothelial damage, neutrophil activation, and thrombosis specific to organoid-derived human blood vessels, as verified through intravital imaging. We examined a new long-acting, pH-sensitive (acid-switched) antibody targeting CFD. In both human and macaque COVID-19 models, this long-acting anti-CFD monoclonal antibody mitigated abnormal complement activation, protected endothelial cells, and curtailed the innate immune response post-viral exposure. Collectively, our findings suggest that the complement alternative pathway exacerbates endothelial injury and inflammation. This underscores the potential of CFD-targeted therapeutics against severe viral-induced inflammathrombotic outcomes.

  Oct. 2023, Cell stem cell, 30(10) (10), 1315 - 1330, English, International magazine

  Scientific journal


• An inhaled ACE2 decoy confers protection against SARS-CoV-2 infection in preclinical models

  Emiko Urano, Yumi Itoh, Tatsuya Suzuki, Takanori Sasaki, Jun Ichi Kishikawa, Kanako Akamatsu, Yusuke Higuchi, Yusuke Sakai, Tomotaka Okamura, Shuya Mitoma, Fuminori Sugihara, Akira Takada, Mari Kimura, Shuto Nakao, Mika Hirose, Tadahiro Sasaki, Ritsuko Koketsu, Shunya Tsuji, Shota Yanagida, Tatsuo Shioda, Eiji Hara, Satoaki Matoba, Yoshiharu Matsuura, Yasunari Kanda, Hisashi Arase, Masato Okada, Junichi Takagi, Takayuki Kato, Atsushi Hoshino, Yasuhiro Yasutomi, Akatsuki Saito, Toru Okamoto

  Aug. 2023, Science translational medicine, 15(711) (711), eadi2623

  Scientific journal


• Combination of HBA1, TTR, and SERPINF2 in plasma defines phenotype correlated with severe burn outcome

  Shinya Onishi, Hisatake Matsumoto, Fuminori Sugihara, Takeshi Ebihara, Hiroshi Matsuura, Akinori Osuka, Daisuke Okuzaki, Hiroshi Ogura, Jun Oda

  Aug. 2023, iScience, 26(8) (8)

  Scientific journal


• 機械学習による新規外傷フェノタイプの同定とプロテオーム解析を用いた病態考察

  舘野 丈太郎, 松本 寿健, 杉原 文徳, 瀬尾 茂人, 奥崎 大介, 北村 哲久, 小向 翔, 木戸 善之, 兒嶋 嵩, 戸上 由貴, 片山 祐介, 中川 雄公, 小倉 裕司, 織田 順

  (一社)日本集中治療医学会, Jun. 2023, 日本集中治療医学会雑誌, 30(Suppl.1) (Suppl.1), S456 - S456, Japanese


• Single-cell analyses and host genetics highlight the role of innate immune cells in COVID-19 severity.

  Ryuya Edahiro, Yuya Shirai, Yusuke Takeshima, Shuhei Sakakibara, Yuta Yamaguchi, Teruaki Murakami, Takayoshi Morita, Yasuhiro Kato, Yu-Chen Liu, Daisuke Motooka, Yoko Naito, Ayako Takuwa, Fuminori Sugihara, Kentaro Tanaka, James B Wing, Kyuto Sonehara, Yoshihiko Tomofuji, Ho Namkoong, Hiromu Tanaka, Ho Lee, Koichi Fukunaga, Haruhiko Hirata, Yoshito Takeda, Daisuke Okuzaki, Atsushi Kumanogoh, Yukinori Okada

  Mechanisms underpinning the dysfunctional immune response in severe acute respiratory syndrome coronavirus 2 infection are elusive. We analyzed single-cell transcriptomes and T and B cell receptors (BCR) of >895,000 peripheral blood mononuclear cells from 73 coronavirus disease 2019 (COVID-19) patients and 75 healthy controls of Japanese ancestry with host genetic data. COVID-19 patients showed a low fraction of nonclassical monocytes (ncMono). We report downregulated cell transitions from classical monocytes to ncMono in COVID-19 with reduced CXCL10 expression in ncMono in severe disease. Cell-cell communication analysis inferred decreased cellular interactions involving ncMono in severe COVID-19. Clonal expansions of BCR were evident in the plasmablasts of patients. Putative disease genes identified by COVID-19 genome-wide association study showed cell type-specific expressions in monocytes and dendritic cells. A COVID-19-associated risk variant at the IFNAR2 locus (rs13050728) had context-specific and monocyte-specific expression quantitative trait loci effects. Our study highlights biological and host genetic involvement of innate immune cells in COVID-19 severity.

  Apr. 2023, Nature genetics, 55(5) (5), 753 - 767, English, International magazine

  Scientific journal


• Proximity-dependent biotin labeling in testicular germ cells identified TESMIN-associated proteins.

  Seiya Oura, Akinori Ninomiya, Fuminori Sugihara, Martin M Matzuk, Masahito Ikawa

  Characterization of protein-protein interactions (PPI) is a key to understanding the functions of proteins of interest. Recently developed proximity-dependent biotin identification (BioID) has been actively investigated as an alternative PPI mapping method because of its usefulness in uncovering transient PPI. Here, as an example of proximity labeling proteomics application in the testis, we generated two transgenic mouse lines expressing two biotin ligases (BioID2 or TurboID) fused with TESMIN, which translocates from the cytosol to the nucleus during meiotic progression and is required for reproduction. The BioID2 transgene, albeit not the TurboID transgene, rescued fertility defects of the Tesmin KO male mice, indicating that the TESMIN-BioID2 fusion can physiologically replace TESMIN. Furthermore, biotinylated protein pull-down and affinity-purification followed by mass spectrometry using the TESMIN-BioID2 transgenic mice captured components of the MYBL1-MuvB complex that regulate cell-cycle gene expression. Thus, our study shows that proximity labeling proteomics can be applied in male germ cells, although the choice of biotin ligase needs to be carefully tested.

  Dec. 2022, Scientific reports, 12(1) (1), 22198 - 22198, English, International magazine

  Scientific journal


• Langerhans cell histiocytosis associated with primary myelofibrosis presenting as disseminated maculopapular rash.

  Moeko Isei, Jun Nakata, Rikako Deno, Fuminori Sugihara, Ai Matsuura, Masaru Shibano, Yumiko Yasuhara, Shin-Ichi Nakatsuka, Aya Tanaka

  Dec. 2022, The Journal of dermatology, 50(4) (4), e133-e134, English, International magazine


• The lysosomal Ragulator complex activates NLRP3 inflammasome in vivo via HDAC6.

  Kohei Tsujimoto, Tatsunori Jo, Daiki Nagira, Hachiro Konaka, Jeong Hoon Park, Shin-Ichiro Yoshimura, Akinori Ninomiya, Fuminori Sugihara, Takehiro Hirayama, Eri Itotagawa, Yusei Matsuzaki, Yuki Takaichi, Wataru Aoki, Shotaro Saita, Shuhei Nakamura, Andrea Ballabio, Shigeyuki Nada, Masato Okada, Hyota Takamatsu, Atsushi Kumanogoh

  The cellular activation of the NLRP3 inflammasome is spatiotemporally orchestrated by various organelles, but whether lysosomes contribute to this process remains unclear. Here, we show the vital role of the lysosomal membrane-tethered Ragulator complex in NLRP3 inflammasome activation. Deficiency of Lamtor1, an essential component of the Ragulator complex, abrogated NLRP3 inflammasome activation in murine macrophages and human monocytic cells. Myeloid-specific Lamtor1-deficient mice showed marked attenuation of NLRP3-associated inflammatory disease severity, including LPS-induced sepsis, alum-induced peritonitis, and monosodium urate (MSU)-induced arthritis. Mechanistically, Lamtor1 interacted with both NLRP3 and histone deacetylase 6 (HDAC6). HDAC6 enhances the interaction between Lamtor1 and NLRP3, resulting in NLRP3 inflammasome activation. DL-all-rac-α-tocopherol, a synthetic form of vitamin E, inhibited the Lamtor1-HDAC6 interaction, resulting in diminished NLRP3 inflammasome activation. Further, DL-all-rac-α-tocopherol alleviated acute gouty arthritis and MSU-induced peritonitis. These results provide novel insights into the role of lysosomes in the activation of NLRP3 inflammasomes by the Ragulator complex.

  Nov. 2022, The EMBO journal, 42(1) (1), e111389, English, International magazine

  Scientific journal


• Transcriptional differences between coronavirus disease 2019 and bacterial sepsis.

  Hiroshi Ito, Masakazu Ishikawa, Hisatake Matsumoto, Fuminori Sugihara, Daisuke Okuzaki, Haruhiko Hirata, Hiroshi Ogura

  BACKGROUND: Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2, has led to major public health crises worldwide. Several studies have reported the comprehensive mRNA expression analysis of immune-related genes in patients with COVID-19, using blood samples, to understand its pathogenesis; however, the characteristics of RNA expression in COVID-19 and bacterial sepsis have not been compared. The current study aimed to address this gap. METHODS: RNA-sequencing and bioinformatics analyses were used to compare the transcriptome expression of whole blood samples from patients with COVID-19 and patients with sepsis who were admitted to the intensive care unit of Osaka University Graduate School of Medicine. RESULTS: The COVID-19 and sepsis cohorts showed upregulation of mitochondrial- and neutrophil-related transcripts, respectively. Compared with that in the control cohort, neutrophil-related transcripts were upregulated in both the COVID-19 and sepsis cohorts. In contrast, mitochondrial-related transcripts were upregulated in the COVID-19 cohort and downregulated in the sepsis cohort, compared to those in the control cohort. Moreover, transcript levels of the pro-apoptotic genes BAK1, CYCS, BBC3, CASP7, and CASP8 were upregulated in the COVID-19 cohort, whereas those of anti-apoptotic genes, such as BCL2L11 and BCL2L1, were upregulated in the sepsis cohort. CONCLUSIONS: This study clarified the differential expression of transcripts related to neutrophils and mitochondria in sepsis and COVID-19 conditions. Mitochondrial-related transcripts were downregulated in sepsis than in COVID-19 conditions, and our results indicated suboptimal intrinsic apoptotic features in sepsis samples compared with that in COVID-19 samples. This study is expected to contribute to the development of specific treatments for COVID-19.

  Nov. 2022, Virology journal, 19(1) (1), 198 - 198, English, International magazine

  Scientific journal


• Combination of WFDC2, CHI3L1, and KRT19 in Plasma Defines a Clinically Useful Molecular Phenotype Associated with Prognosis in Critically Ill COVID-19 Patients.

  Takeshi Ebihara, Tsunehiro Matsubara, Yuki Togami, Hisatake Matsumoto, Jotaro Tachino, Hiroshi Matsuura, Takashi Kojima, Fuminori Sugihara, Shigeto Seno, Daisuke Okuzaki, Haruhiko Hirata, Hiroshi Ogura

  BACKGROUND: COVID-19 is now a common disease, but its pathogenesis remains unknown. Blood circulating proteins reflect host defenses against COVID-19. We investigated whether evaluation of longitudinal blood proteomics for COVID-19 and merging with clinical information would allow elucidation of its pathogenesis and develop a useful clinical phenotype. METHODS: To achieve the first goal (determining key proteins), we derived plasma proteins related to disease severity by using a first discovery cohort. We then assessed the association of the derived proteins with clinical outcome in a second discovery cohort. Finally, the candidates were validated by enzyme-linked immunosorbent assay in a validation cohort to determine key proteins. For the second goal (understanding the associations of the clinical phenotypes with 28-day mortality and clinical outcome), we assessed the associations between clinical phenotypes derived by latent cluster analysis with the key proteins and 28-day mortality and clinical outcome. RESULTS: We identified four key proteins (WFDC2, GDF15, CHI3L1, and KRT19) involved in critical pathogenesis from the three different cohorts. These key proteins were related to the function of cell adhesion and not immune response. Considering the multicollinearity, three clinical phenotypes based on WFDC2, CHI3L1, and KRT19 were identified that were associated with mortality and clinical outcome. CONCLUSION: The use of these easily measured key proteins offered new insight into the pathogenesis of COVID-19 and could be useful in a potential clinical application.

  Nov. 2022, Journal of clinical immunology, 43(2) (2), 1 - 13, English, International magazine

  Scientific journal


• DOCK2 is involved in the host genetics and biology of severe COVID-19.

  Ho Namkoong, Ryuya Edahiro, Tomomi Takano, Hiroshi Nishihara, Yuya Shirai, Kyuto Sonehara, Hiromu Tanaka, Shuhei Azekawa, Yohei Mikami, Ho Lee, Takanori Hasegawa, Koji Okudela, Daisuke Okuzaki, Daisuke Motooka, Masahiro Kanai, Tatsuhiko Naito, Kenichi Yamamoto, Qingbo S Wang, Ryunosuke Saiki, Rino Ishihara, Yuta Matsubara, Junko Hamamoto, Hiroyuki Hayashi, Yukihiro Yoshimura, Natsuo Tachikawa, Emmy Yanagita, Takayoshi Hyugaji, Eigo Shimizu, Kotoe Katayama, Yasuhiro Kato, Takayoshi Morita, Kazuhisa Takahashi, Norihiro Harada, Toshio Naito, Makoto Hiki, Yasushi Matsushita, Haruhi Takagi, Ryousuke Aoki, Ai Nakamura, Sonoko Harada, Hitoshi Sasano, Hiroki Kabata, Katsunori Masaki, Hirofumi Kamata, Shinnosuke Ikemura, Shotaro Chubachi, Satoshi Okamori, Hideki Terai, Atsuho Morita, Takanori Asakura, Junichi Sasaki, Hiroshi Morisaki, Yoshifumi Uwamino, Kosaku Nanki, Sho Uchida, Shunsuke Uno, Tomoyasu Nishimura, Takashi Ishiguro, Taisuke Isono, Shun Shibata, Yuma Matsui, Chiaki Hosoda, Kenji Takano, Takashi Nishida, Yoichi Kobayashi, Yotaro Takaku, Noboru Takayanagi, Soichiro Ueda, Ai Tada, Masayoshi Miyawaki, Masaomi Yamamoto, Eriko Yoshida, Reina Hayashi, Tomoki Nagasaka, Sawako Arai, Yutaro Kaneko, Kana Sasaki, Etsuko Tagaya, Masatoshi Kawana, Ken Arimura, Kunihiko Takahashi, Tatsuhiko Anzai, Satoshi Ito, Akifumi Endo, Yuji Uchimura, Yasunari Miyazaki, Takayuki Honda, Tomoya Tateishi, Shuji Tohda, Naoya Ichimura, Kazunari Sonobe, Chihiro Tani Sassa, Jun Nakajima, Yasushi Nakano, Yukiko Nakajima, Ryusuke Anan, Ryosuke Arai, Yuko Kurihara, Yuko Harada, Kazumi Nishio, Tetsuya Ueda, Masanori Azuma, Ryuichi Saito, Toshikatsu Sado, Yoshimune Miyazaki, Ryuichi Sato, Yuki Haruta, Tadao Nagasaki, Yoshinori Yasui, Yoshinori Hasegawa, Yoshikazu Mutoh, Tomoki Kimura, Tomonori Sato, Reoto Takei, Satoshi Hagimoto, Yoichiro Noguchi, Yasuhiko Yamano, Hajime Sasano, Sho Ota, Yasushi Nakamori, Kazuhisa Yoshiya, Fukuki Saito, Tomoyuki Yoshihara, Daiki Wada, Hiromu Iwamura, Syuji Kanayama, Shuhei Maruyama, Takashi Yoshiyama, Ken Ohta, Hiroyuki Kokuto, Hideo Ogata, Yoshiaki Tanaka, Kenichi Arakawa, Masafumi Shimoda, Takeshi Osawa, Hiroki Tateno, Isano Hase, Shuichi Yoshida, Shoji Suzuki, Miki Kawada, Hirohisa Horinouchi, Fumitake Saito, Keiko Mitamura, Masao Hagihara, Junichi Ochi, Tomoyuki Uchida, Rie Baba, Daisuke Arai, Takayuki Ogura, Hidenori Takahashi, Shigehiro Hagiwara, Genta Nagao, Shunichiro Konishi, Ichiro Nakachi, Koji Murakami, Mitsuhiro Yamada, Hisatoshi Sugiura, Hirohito Sano, Shuichiro Matsumoto, Nozomu Kimura, Yoshinao Ono, Hiroaki Baba, Yusuke Suzuki, Sohei Nakayama, Keita Masuzawa, Shinichi Namba, Ken Suzuki, Yoko Naito, Yu-Chen Liu, Ayako Takuwa, Fuminori Sugihara, James B Wing, Shuhei Sakakibara, Nobuyuki Hizawa, Takayuki Shiroyama, Satoru Miyawaki, Yusuke Kawamura, Akiyoshi Nakayama, Hirotaka Matsuo, Yuichi Maeda, Takuro Nii, Yoshimi Noda, Takayuki Niitsu, Yuichi Adachi, Takatoshi Enomoto, Saori Amiya, Reina Hara, Yuta Yamaguchi, Teruaki Murakami, Tomoki Kuge, Kinnosuke Matsumoto, Yuji Yamamoto, Makoto Yamamoto, Midori Yoneda, Toshihiro Kishikawa, Shuhei Yamada, Shuhei Kawabata, Noriyuki Kijima, Masatoshi Takagaki, Noah Sasa, Yuya Ueno, Motoyuki Suzuki, Norihiko Takemoto, Hirotaka Eguchi, Takahito Fukusumi, Takao Imai, Munehisa Fukushima, Haruhiko Kishima, Hidenori Inohara, Kazunori Tomono, Kazuto Kato, Meiko Takahashi, Fumihiko Matsuda, Haruhiko Hirata, Yoshito Takeda, Hidefumi Koh, Tadashi Manabe, Yohei Funatsu, Fumimaro Ito, Takahiro Fukui, Keisuke Shinozuka, Sumiko Kohashi, Masatoshi Miyazaki, Tomohisa Shoko, Mitsuaki Kojima, Tomohiro Adachi, Motonao Ishikawa, Kenichiro Takahashi, Takashi Inoue, Toshiyuki Hirano, Keigo Kobayashi, Hatsuyo Takaoka, Kazuyoshi Watanabe, Naoki Miyazawa, Yasuhiro Kimura, Reiko Sado, Hideyasu Sugimoto, Akane Kamiya, Naota Kuwahara, Akiko Fujiwara, Tomohiro Matsunaga, Yoko Sato, Takenori Okada, Yoshihiro Hirai, Hidetoshi Kawashima, Atsuya Narita, Kazuki Niwa, Yoshiyuki Sekikawa, Koichi Nishi, Masaru Nishitsuji, Mayuko Tani, Junya Suzuki, Hiroki Nakatsumi, Takashi Ogura, Hideya Kitamura, Eri Hagiwara, Kota Murohashi, Hiroko Okabayashi, Takao Mochimaru, Shigenari Nukaga, Ryosuke Satomi, Yoshitaka Oyamada, Nobuaki Mori, Tomoya Baba, Yasutaka Fukui, Mitsuru Odate, Shuko Mashimo, Yasushi Makino, Kazuma Yagi, Mizuha Hashiguchi, Junko Kagyo, Tetsuya Shiomi, Satoshi Fuke, Hiroshi Saito, Tomoya Tsuchida, Shigeki Fujitani, Mumon Takita, Daiki Morikawa, Toru Yoshida, Takehiro Izumo, Minoru Inomata, Naoyuki Kuse, Nobuyasu Awano, Mari Tone, Akihiro Ito, Yoshihiko Nakamura, Kota Hoshino, Junichi Maruyama, Hiroyasu Ishikura, Tohru Takata, Toshio Odani, Masaru Amishima, Takeshi Hattori, Yasuo Shichinohe, Takashi Kagaya, Toshiyuki Kita, Kazuhide Ohta, Satoru Sakagami, Kiyoshi Koshida, Kentaro Hayashi, Tetsuo Shimizu, Yutaka Kozu, Hisato Hiranuma, Yasuhiro Gon, Namiki Izumi, Kaoru Nagata, Ken Ueda, Reiko Taki, Satoko Hanada, Kodai Kawamura, Kazuya Ichikado, Kenta Nishiyama, Hiroyuki Muranaka, Kazunori Nakamura, Naozumi Hashimoto, Keiko Wakahara, Sakamoto Koji, Norihito Omote, Akira Ando, Nobuhiro Kodama, Yasunari Kaneyama, Shunsuke Maeda, Takashige Kuraki, Takemasa Matsumoto, Koutaro Yokote, Taka-Aki Nakada, Ryuzo Abe, Taku Oshima, Tadanaga Shimada, Masahiro Harada, Takeshi Takahashi, Hiroshi Ono, Toshihiro Sakurai, Takayuki Shibusawa, Yoshifumi Kimizuka, Akihiko Kawana, Tomoya Sano, Chie Watanabe, Ryohei Suematsu, Hisako Sageshima, Ayumi Yoshifuji, Kazuto Ito, Saeko Takahashi, Kota Ishioka, Morio Nakamura, Makoto Masuda, Aya Wakabayashi, Hiroki Watanabe, Suguru Ueda, Masanori Nishikawa, Yusuke Chihara, Mayumi Takeuchi, Keisuke Onoi, Jun Shinozuka, Atsushi Sueyoshi, Yoji Nagasaki, Masaki Okamoto, Sayoko Ishihara, Masatoshi Shimo, Yoshihisa Tokunaga, Yu Kusaka, Takehiko Ohba, Susumu Isogai, Aki Ogawa, Takuya Inoue, Satoru Fukuyama, Yoshihiro Eriguchi, Akiko Yonekawa, Keiko Kan-O, Koichiro Matsumoto, Kensuke Kanaoka, Shoichi Ihara, Kiyoshi Komuta, Yoshiaki Inoue, Shigeru Chiba, Kunihiro Yamagata, Yuji Hiramatsu, Hirayasu Kai, Koichiro Asano, Tsuyoshi Oguma, Yoko Ito, Satoru Hashimoto, Masaki Yamasaki, Yu Kasamatsu, Yuko Komase, Naoya Hida, Takahiro Tsuburai, Baku Oyama, Minoru Takada, Hidenori Kanda, Yuichiro Kitagawa, Tetsuya Fukuta, Takahito Miyake, Shozo Yoshida, Shinji Ogura, Shinji Abe, Yuta Kono, Yuki Togashi, Hiroyuki Takoi, Ryota Kikuchi, Shinichi Ogawa, Tomouki Ogata, Shoichiro Ishihara, Arihiko Kanehiro, Shinji Ozaki, Yasuko Fuchimoto, Sae Wada, Nobukazu Fujimoto, Kei Nishiyama, Mariko Terashima, Satoru Beppu, Kosuke Yoshida, Osamu Narumoto, Hideaki Nagai, Nobuharu Ooshima, Mitsuru Motegi, Akira Umeda, Kazuya Miyagawa, Hisato Shimada, Mayu Endo, Yoshiyuki Ohira, Masafumi Watanabe, Sumito Inoue, Akira Igarashi, Masamichi Sato, Hironori Sagara, Akihiko Tanaka, Shin Ohta, Tomoyuki Kimura, Yoko Shibata, Yoshinori Tanino, Takefumi Nikaido, Hiroyuki Minemura, Yuki Sato, Yuichiro Yamada, Takuya Hashino, Masato Shinoki, Hajime Iwagoe, Hiroshi Takahashi, Kazuhiko Fujii, Hiroto Kishi, Masayuki Kanai, Tomonori Imamura, Tatsuya Yamashita, Masakiyo Yatomi, Toshitaka Maeno, Shinichi Hayashi, Mai Takahashi, Mizuki Kuramochi, Isamu Kamimaki, Yoshiteru Tominaga, Tomoo Ishii, Mitsuyoshi Utsugi, Akihiro Ono, Toru Tanaka, Takeru Kashiwada, Kazue Fujita, Yoshinobu Saito, Masahiro Seike, Hiroko Watanabe, Hiroto Matsuse, Norio Kodaka, Chihiro Nakano, Takeshi Oshio, Takatomo Hirouchi, Shohei Makino, Moritoki Egi, Yosuke Omae, Yasuhito Nannya, Takafumi Ueno, Kazuhiko Katayama, Masumi Ai, Yoshinori Fukui, Atsushi Kumanogoh, Toshiro Sato, Naoki Hasegawa, Katsushi Tokunaga, Makoto Ishii, Ryuji Koike, Yuko Kitagawa, Akinori Kimura, Seiya Imoto, Satoru Miyano, Seishi Ogawa, Takanori Kanai, Koichi Fukunaga, Yukinori Okada

  Identifying the factors underlying severe COVID-19 in the host genetics is an emerging issue1-5. We conducted a genome-wide association study (GWAS) involving 2,393 Japanese COVID-19 cases collected in initial pandemic waves with 3,289 controls, which identified a variant on 5q35 (rs60200309-A) near DOCK2 associated with severe COVID-19 in younger (<65 ages) patients (nCase=440, odds ratio=2.01, P=1.2×10-8). This risk allele was prevalent in East Asians but rare in Europeans, showing a value of non-European GWAS. RNA-seq of 473 bulk peripheral blood identified decreasing effect of the risk allele on DOCK2 expression in younger patients. DOCK2 expression was suppressed in severe forms of COVID-19. Single cell RNA-seq analysis (n=61) identified cell type-specific downregulation of DOCK2 and COVID-19-specific decreasing effects of the risk allele on DOCK2 in non-classical monocytes. Immunohistochemistry of lung specimens from severe COVID-19 pneumonia showed suppressed DOCK2. Moreover, inhibition of DOCK2 function using CPYPP induced much more severe pneumonia in a Syrian hamster model of SARS-CoV-2 infection characterized as weight loss, lung edema, enhanced viral loads, impaired macrophage recruitment and dysregulated type I interferon responses. We conclude that DOCK2 plays an important role in the host immune response to SARS-CoV-2 infection and development of severe COVID-19, and could be further explored as a potential biomarker and/or therapeutic target.

  Aug. 2022, Nature, 609(7928) (7928), 754 - 760, English, International magazine

  Scientific journal


• Development of clinical phenotypes and biological profiles via proteomic analysis of trauma patients

  Jotaro Tachino, Hisatake Matsumoto, Fuminori Sugihara, Shigeto Seno, Daisuke Okuzaki, Tetsuhisa Kitamura, Sho Komukai, Yoshiyuki Kido, Takashi Kojima, Yuki Togami, Yusuke Katayama, Yuko Nakagawa, Hiroshi Ogura

  Abstract Background Trauma is a heterogeneous condition, and specific clinical phenotypes may identify target populations that could benefit from certain treatment strategies. In this retrospective study, we determined clinical phenotypes and identified new target populations of trauma patients and their treatment strategies. Methods We retrospectively analyzed datasets from the Japan Trauma Data Bank and determined trauma death clinical phenotypes using statistical machine learning techniques and evaluation of biological profiles. Results The analysis included 71,038 blunt trauma patients [median age, 63 (interquartile range [IQR], 40–78) years; 45,479 (64.0%) males; median Injury Severity Score, 13 (IQR, 9–20)], and the derivation and validation cohorts included 42,780 (60.2%) and 28,258 (39.8%) patients, respectively. Of eight derived phenotypes (D-1–D-8), D-8 (n = 2178) had the highest mortality (48.6%) with characteristic severely disturbed consciousness and was further divided into four phenotypes: D-8α, multiple trauma in the young (n = 464); D-8β, head trauma with lower body temperature (n = 178); D-8γ, severe head injury in the elderly (n = 957); and D-8δ, multiple trauma, with higher predicted mortality than actual mortality (n = 579). Phenotype distributions were comparable in the validation cohort. Biological profile analysis of 90 trauma patients revealed that D-8 exhibited excessive inflammation, including enhanced acute inflammatory response, dysregulated complement activation pathways, and impaired coagulation, including downregulated coagulation and platelet degranulation pathways, compared with other phenotypes. Conclusions We identified clinical phenotypes with high mortality, and the evaluation of the molecular pathogenesis underlying these clinical phenotypes suggests that lethal trauma may involve excessive inflammation and coagulation disorders.

  Springer Science and Business Media LLC, Aug. 2022, Critical Care, 26(1) (1), 241 - 241, English, International magazine

  Scientific journal


• Clinical Significance of Interferon Signaling Based on mRNA-microRNA Integration and Plasma Protein Analyses in Critically Ill COVID-19 Patients.

  Yuki Togami, Hisatake Matsumoto, Jumpei Yoshimura, Tsunehiro Matsubara, Takeshi Ebihara, Hiroshi Matsuura, Yumi Mitsuyama, Takashi Kojima, Masakazu Ishikawa, Fuminori Sugihara, Haruhiko Hirata, Daisuke Okuzaki, Hiroshi Ogura

  We evaluated mRNA and miRNA in COVID-19 patients and elucidated the pathogenesis of COVID-19 including protein profiles following mRNA and miRNA integration analysis. mRNA and miRNA sequencing was done on admission with whole blood of 5 and 16 healthy controls (HC) and 10 and 31 critically ill COVID-19 patients (derivation and validation cohorts, respectively). Interferon (IFN)-α2, IFN-β, IFN-γ, interleukin-27, and IFN-λ1 were measured in COVID-19 patients on admission (day 1, 181 critical/22 non-critical patients) and days 6-8 (168 critical patients) and in 19 HC. In the derivation cohort, 3488 mRNA and 31 miRNA expressions were identified among differentially expressed RNA expressions in the patients versus those in HC, and 2945 mRNA and 32 miRNA expressions in the validation cohort. Canonical pathway analysis showed the IFN signaling pathway to be most activated. The IFN-β plasma level was elevated in line with increased severity compared to HC, as were IFN-β downstream proteins such as interleukin-27. IFN-λ1 was higher in non-critically ill patients versus HC but lower in critical than non-critical patients. Integration of mRNA and miRNA analysis showed activated IFN signaling. Plasma IFN proteins profile revealed that IFN-β (type I) and IFN-λ1 (type III) played important roles in COVID-19 disease progression.

  Jul. 2022, Molecular therapy. Nucleic acids, 29, 343 - 353, English, International magazine

  Scientific journal


• Humanized Anti-RGMa Antibody Treatment Promotes Repair of Blood-Spinal Cord Barrier Under Autoimmune Encephalomyelitis in Mice.

  Takeshi Hirata, Takahide Itokazu, Atsushi Sasaki, Fuminori Sugihara, Toshihide Yamashita

  The lack of established biomarkers which reflect dynamic neuropathological alterations in multiple sclerosis (MS) makes it difficult to determine the therapeutic response to the tested drugs and to identify the key biological process that mediates the beneficial effect of them. In the present study, we applied high-field MR imaging in locally-induced experimental autoimmune encephalomyelitis (EAE) mice to evaluate dynamic changes following treatment with a humanized anti-repulsive guidance molecule-a (RGMa) antibody, a potential drug for MS. Based on the longitudinal evaluation of various MRI parameters including white matter, axon, and myelin integrity as well as blood-spinal cord barrier (BSCB) disruption, anti-RGMa antibody treatment exhibited a strong and prompt therapeutic effect on the disrupted BSCB, which was paralleled by functional improvement. The antibody's effect on BSCB repair was also suggested via GeneChip analysis. Moreover, immunohistochemical analysis revealed that EAE-induced vascular pathology which is characterized by aberrant thickening of endothelial cells and perivascular type I/IV collagen deposits were attenuated by anti-RGMa antibody treatment, further supporting the idea that the BSCB is one of the key therapeutic targets of anti-RGMa antibody. Importantly, the extent of BSCB disruption detected by MRI could predict late-phase demyelination, and the predictability of myelin integrity based on the extent of acute-phase BSCB disruption was compromised following anti-RGMa antibody treatment. These results strongly support the concept that longitudinal MRI with simultaneous DCE-MRI and DTI analysis can be used as an imaging biomarker and is useful for unbiased prioritization of the key biological process that mediates the therapeutic effect of tested drugs.

  Jun. 2022, Frontiers in immunology, 13, 870126 - 870126, English, International magazine

  [Refereed]

  Scientific journal


• Resistin Associated With Cytokines and Endothelial Cell Adhesion Molecules Is Related to Worse Outcome in COVID-19.

  Takeshi Ebihara, Hisatake Matsumoto, Tsunehiro Matsubara, Yuki Togami, Shunichiro Nakao, Hiroshi Matsuura, Shinya Onishi, Takashi Kojima, Fuminori Sugihara, Daisuke Okuzaki, Haruhiko Hirata, Hitoshi Yamamura, Hiroshi Ogura

  Introduction: Resistin is reported to form a cytokine network and cause endothelial damage. The pathogenesis of coronavirus disease 2019 (COVID-19) remains unknown, but the association between cytokine storm and endothelial damage is crucial. This study aimed to evaluate resistin in COVID-19 pathogenesis compared with sepsis. Materials and Methods: First, we evaluated the association of plasma resistin levels and disease severity and clinical outcome in two large cohorts: a publicly available cohort including 306 COVID-19 patients in the United States (MGH cohort) and our original cohort including only intubated 113 patients in Japan (Osaka cohort 1). Second, to understand pathogenesis, we evaluate resistin, cytokines and endothelial cell adhesion molecules in COVID-19 compared with sepsis. Blood samples were collected from 62 ICU-treated COVID-19 patients and 38 sepsis patients on day 1 (day of ICU admission), days 2-3, days 6-8, and from 18 healthy controls (Osaka cohort 2). The plasma resistin, inflammatory cytokines (IL-6, IL-8, MCP-1 and IL-10) and endothelial cell adhesion molecules (ICAM-1 and VCAM-1) were compared between patients and control. Correlations among resistin, inflammatory cytokines and endothelial cell adhesion molecules were evaluated in COVID-19 and sepsis. Results: In the MGH cohort, the day 1 resistin levels were associated with disease severity score. The non-survivors showed significantly greater resistin levels than survivors on days 1, 4 and 8. In the Osaka cohort 1, 28-day non-survivors showed significantly higher resistin levels than 28-day survivors on days 6-8. Patients with late recovery (defined as the day of weaning off mechanical ventilation >12 or death) had significantly higher resistin levels than those with early recovery on day 1 and days 6-8. In the Osaka cohort 2, plasma resistin levels were elevated in COVID-19 and sepsis patients compared to controls at all measurement points and were associated with inflammatory cytokines and endothelial cell adhesion molecules. Conclusion: Resistin was elevated in COVID-19 patients and was associated with cytokines and endothelial cell adhesion molecules. Higher resistin levels were related to worse outcome.

  Jun. 2022, Frontiers in immunology, 13, 830061 - 830061, English, International magazine

  [Refereed]

  Scientific journal


• A spontaneous pulmonary fibrosis mouse model identifies a mechanism for ILC2-mediated irreversible fibrosis

  Kazuyo Moro, Natsuko Otaki, Yasutaka Motomura, Tommy Terooatea, S. Thomas Kelly, Miho Mochizuki, Natsuki Takeno, Shigeo Koyasu, Fuminori Sugihara, Junichi Kikuta, Hideya Kitamura, Yoshiki Shiraishi, Jun Miyanohara, Yuji Nagano, Yuji Saita, Takashi Ogura, Koichiro Asano, Aki Minoda

  Abstract Pulmonary fibrosis (PF) is characterised by inflammation and collagen deposition in the alveolar interstitium, leading to dyspnoea and death. However, the comprehensive pathogenesis of PF remains unclear due to the lack of spontaneous fibrosis mouse models. Here, we found that Ifngr1^-/-Rag2^-/- mice, lacking the mechanisms to suppress group 2 and 3 innate lymphoid cells (ILC2s and ILC3s), developed severe PF spontaneously. In Ifngr1^-/-Rag2^-/- mice, Il1rl1^hiIl13^hi-ILC2 subpopulation was increased at disease-onset phase before collagen production began. Further, defects in ILCs or IL-33, the strong ILC2 activator, prevented PF development. ILC2s directly induce collagen production by fibroblasts in vitro, and fibroblasts started to produce IL-33 in the chronic phase, presumably forming a positive feedback loop between fibroblasts and ILC2s leading to irreversible fibrosis. Moreover, the increased IL1RL1 and IL13 and decreased IFNGR1 expression levels in ILC2s from human idiopathic PF patients highlight the significance of the novel mouse model in fibrosis research.

  Research Square Platform LLC, May 2022


• pH‐Sensitive Polymethacrylates as Potential Contrast Agents in ¹⁹ F MRI

  Mariusz Zalewski, Dawid Janasik, Anna Kapała, Masafumi Minoshima, Fuminori Sugihara, Wojciech Raj, Joanna Pietrasik, Kazuya Kikuchi, Tomasz Krawczyk

  Wiley, Apr. 2022, Macromolecular Chemistry and Physics, 223(14) (14), 2200027 - 2200027

  Scientific journal


• Osteoblast-derived vesicles induce a switch from bone-formation to bone-resorption in vivo.

  Maki Uenaka, Erika Yamashita, Junichi Kikuta, Akito Morimoto, Tomoka Ao, Hiroki Mizuno, Masayuki Furuya, Tetsuo Hasegawa, Hiroyuki Tsukazaki, Takao Sudo, Keizo Nishikawa, Daisuke Okuzaki, Daisuke Motooka, Nobuyoshi Kosaka, Fuminori Sugihara, Thomas Boettger, Thomas Braun, Takahiro Ochiya, Masaru Ishii

  Bone metabolism is regulated by the cooperative activity between bone-forming osteoblasts and bone-resorbing osteoclasts. However, the mechanisms mediating the switch between the osteoblastic and osteoclastic phases have not been fully elucidated. Here, we identify a specific subset of mature osteoblast-derived extracellular vesicles that inhibit bone formation and enhance osteoclastogenesis. Intravital imaging reveals that mature osteoblasts secrete and capture extracellular vesicles, referred to as small osteoblast vesicles (SOVs). Co-culture experiments demonstrate that SOVs suppress osteoblast differentiation and enhance the expression of receptor activator of NF-κB ligand, thereby inducing osteoclast differentiation. We also elucidate that the SOV-enriched microRNA miR-143 inhibits Runt-related transcription factor 2, a master regulator of osteoblastogenesis, by targeting the mRNA expression of its dimerization partner, core-binding factor β. In summary, we identify SOVs as a mode of cell-to-cell communication, controlling the dynamic transition from bone-forming to bone-resorbing phases in vivo.

  Feb. 2022, Nature communications, 13(1) (1), 1066 - 1066, English, International magazine

  [Refereed]

  Scientific journal


• Cytokine Elevation in Severe COVID-19 From Longitudinal Proteomics Analysis: Comparison With Sepsis.

  Takeshi Ebihara, Hisatake Matsumoto, Tsunehiro Matsubara, Yuki Togami, Shunichiro Nakao, Hiroshi Matsuura, Takashi Kojima, Fuminori Sugihara, Daisuke Okuzaki, Haruhiko Hirata, Hitoshi Yamamura, Hiroshi Ogura

  Introduction: Coronavirus disease 2019 (COVID-19) is a new viral disease. Uncontrolled inflammation called "cytokine storm" is reported to contribute to disease pathogenesis as well as sepsis. We aimed to identify cytokines related to the pathogenesis of COVID-19 through a proteomics analysis of 1463 plasma proteins, validate these cytokines, and compare them with sepsis. Materials and Methods: In a derivation cohort of 306 patients with COVID-19, 1463 unique plasma proteins were measured on days 1, 4, and 8. Cytokines associated with disease severity and prognosis were derived. In a validation cohort of 62 COVID-19 patients and 38 sepsis patients treated in the intensive care unit [ICU], these derived cytokines were measured on days 1 (day of ICU admission), 2-3, and 6-8 (maximum: 3 time points/patient). Derived cytokines were compared with healthy controls and between COVID-19 and sepsis patients, and the associations with prognosis were evaluated. The time to wean off mechanical ventilation (MV) was evaluated only for COVID-19. Results: IL-6, amphiregulin, and growth differentiation factor (GDF)-15 were associated with disease severity and prognosis in the derivation cohort. In the validation cohort, IL-6 and GDF-15 were elevated in COVID-19 and sepsis on day 1, and the levels of these cytokines were higher in sepsis than in COVID-19. IL-6 and GDF-15 were associated with prognosis in sepsis. Cox proportional hazards model with time as a dependent covariate showed a significant relationship between plasma GDF-15 level and time to wean off MV (hazard ratio, 0.549 [95% confidence level, 0.382-0.789]). The GDF-15 level at ICU admission predicted late recovery. Conclusion: GDF-15 and IL-6 derived from proteomics analysis were related with disease severity of COVID-19. Their values were higher in sepsis than in COVID-19 and were associated with prognosis in sepsis. In COVID-19 patients treated in the ICU, GDF-15 was associated with the time to wean off MV and better predicted late recovery.

  Jan. 2022, Frontiers in immunology, 12, 798338 - 798338, English, International magazine

  [Refereed]

  Scientific journal


• Advances in antibody discovery from human BCR repertoires.

  Zichang Xu, Hendra S Ismanto, Hao Zhou, Dianita S Saputri, Fuminori Sugihara, Daron M Standley

  Antibodies make up an important and growing class of compounds used for the diagnosis or treatment of disease. While traditional antibody discovery utilized immunization of animals to generate lead compounds, technological innovations have made it possible to search for antibodies targeting a given antigen within the repertoires of B cells in humans. Here we group these innovations into four broad categories: cell sorting allows the collection of cells enriched in specificity to one or more antigens; BCR sequencing can be performed on bulk mRNA, genomic DNA or on paired (heavy-light) mRNA; BCR repertoire analysis generally involves clustering BCRs into specificity groups or more in-depth modeling of antibody-antigen interactions, such as antibody-specific epitope predictions; validation of antibody-antigen interactions requires expression of antibodies, followed by antigen binding assays or epitope mapping. Together with innovations in Deep learning these technologies will contribute to the future discovery of diagnostic and therapeutic antibodies directly from humans.

  2022, Frontiers in bioinformatics, 2, 1044975 - 1044975, English, International magazine

  Scientific journal


• Melanin Produced by Bordetella parapertussis Confers a Survival Advantage to the Bacterium during Host Infection.

  Yukihiro Hiramatsu, Takashi Nishida, Dendi Krisna Nugraha, Fuminori Sugihara, Yasuhiko Horiguchi

  Bordetella parapertussis causes respiratory infection in humans, with a mild pertussis (whooping cough)-like disease. The organism produces a brown pigment, the nature and biological significance of which have not been elucidated. Here, by screening a transposon library, we demonstrate that the gene encoding 4-hydroxyphenylpyruvate dioxygenase (HppD) is responsible for production of this pigment. Our results also indicate that the brown pigment produced by the bacterium is melanin, because HppD is involved in the biosynthesis of a type of melanin called pyomelanin, and homogentisic acid, the monomeric precursor of pyomelanin, was detected by high-performance liquid chromatography-mass spectrometry analyses. In an infection assay using macrophages, the hppD-deficient mutant was internalized by THP-1 macrophage-like cells, similar to the wild-type strain, but was less able to survive within the cells, indicating that melanin protects B. parapertussis from intracellular killing in macrophages. Mouse infection experiments also showed that the hppD-deficient mutant was eliminated from the respiratory tract more rapidly than the wild-type strain, although the initial colonization levels were comparable between the two strains. In addition, melanin production by B. parapertussis was not regulated by the BvgAS two-component system, which is the master regulator for the expression of genes contributing to the bacterial infection. Taken together, our findings indicate that melanin produced by B. parapertussis in a BvgAS-independent manner confers a survival advantage to the bacterium during host infection. IMPORTANCE In addition to the Gram-negative bacterium Bordetella pertussis, the etiological agent of pertussis, Bordetella parapertussis also causes respiratory infection in humans, with a mild pertussis-like disease. These bacteria are genetically closely related and share many virulence factors, including adhesins and toxins. However, B. parapertussis is clearly distinguished from B. pertussis by its brown pigment production, the bacteriological significance of which remains unclear. Here, we demonstrate that this pigment is melanin, which is known to be produced by a wide range of organisms from prokaryotes to humans and helps the organisms to survive under various environmental stress conditions. Our results show that melanin confers a survival advantage to B. parapertussis within human macrophages through its protective effect against reactive oxygen species and eventually contributes to respiratory infection of the bacterium in mice. This study proposes melanin as a virulence factor involved in the increased survival of B. parapertussis during host infection.

  Oct. 2021, mSphere, 6(5) (5), e0081921, English, International magazine

  Scientific journal


• A sublethal ATP11A mutation associated with neurological deterioration causes aberrant phosphatidylcholine flipping in plasma membranes.

  Katsumori Segawa, Atsuo Kikuchi, Tomoyasu Noji, Yuki Sugiura, Keita Hiraga, Chigure Suzuki, Kazuhiro Haginoya, Yasuko Kobayashi, Mitsuhiro Matsunaga, Yuki Ochiai, Kyoko Yamada, Takuo Nishimura, Shinya Iwasawa, Wataru Shoji, Fuminori Sugihara, Kohei Nishino, Hidetaka Kosako, Masahito Ikawa, Yasuo Uchiyama, Makoto Suematsu, Hiroshi Ishikita, Shigeo Kure, Shigekazu Nagata

  ATP11A translocates phosphatidylserine (PtdSer), but not phosphatidylcholine (PtdCho), from the outer to the inner leaflet of plasma membranes, thereby maintaining the asymmetric distribution of PtdSer. Here, we detected a de novo heterozygous point mutation of ATP11A in a patient with developmental delays and neurological deterioration. Mice carrying the corresponding mutation died perinatally of neurological disorders. This mutation caused an amino acid substitution (Q84E) in the first transmembrane segment of ATP11A, and mutant ATP11A flipped PtdCho. Molecular dynamics simulations revealed that the mutation allowed PtdCho binding at the substrate entry site. Aberrant PtdCho flipping markedly decreased the concentration of PtdCho in the outer leaflet of plasma membranes, whereas sphingomyelin (SM) concentrations in the outer leaflet increased. This change in the distribution of phospholipids altered cell characteristics, including cell growth, cholesterol homeostasis, and sensitivity to sphingomyelinase. Matrix-assisted laser desorption ionization-imaging mass spectrometry (MALDI-IMS) showed a marked increase of SM levels in the brains of Q84E-knockin mouse embryos. These results provide insights into the physiological importance of the substrate specificity of plasma membrane flippases for the proper distribution of PtdCho and SM.

  Sep. 2021, The Journal of clinical investigation, 131(18) (18), English, International magazine

  Scientific journal


• Loss of FCHSD1 leads to amelioration of chronic obstructive pulmonary disease.

  Takahiro Kawasaki, Fuminori Sugihara, Kiyoharu Fukushima, Takanori Matsuki, Hiroshi Nabeshima, Tomohisa Machida, Yuichi Mitsui, Saki Fujimura, Rio Sagawa, Lee Gaheun, Kanako Kuniyoshi, Hiroki Tanaka, Masashi Narazaki, Atsushi Kumanogoh, Shizuo Akira, Takashi Satoh

  Jun. 2021, Proceedings of the National Academy of Sciences of the United States of America, 118(26) (26), English, International magazine

  Scientific journal


• Engineered ACE2 receptor therapy overcomes mutational escape of SARS-CoV-2.

  Yusuke Higuchi, Tatsuya Suzuki, Takao Arimori, Nariko Ikemura, Emiko Mihara, Yuhei Kirita, Eriko Ohgitani, Osam Mazda, Daisuke Motooka, Shota Nakamura, Yusuke Sakai, Yumi Itoh, Fuminori Sugihara, Yoshiharu Matsuura, Satoaki Matoba, Toru Okamoto, Junichi Takagi, Atsushi Hoshino

  Jun. 2021, Nature communications, 12(1) (1), 3802 - 3802, English, International magazine

  Scientific journal


• Development of Off-On Switching 19FMRI Probes for Cathepsin K Activity Detection

  Jun. 2021, Bulletin of the Chemical Society of Japan, 94(6) (6), 1690 - 1694

  Scientific journal


• SLPI is a critical mediator that controls PTH-induced bone formation.

  Akito Morimoto, Junichi Kikuta, Keizo Nishikawa, Takao Sudo, Maki Uenaka, Masayuki Furuya, Tetsuo Hasegawa, Kunihiko Hashimoto, Hiroyuki Tsukazaki, Shigeto Seno, Akira Nakamura, Daisuke Okuzaki, Fuminori Sugihara, Akinori Ninomiya, Takeshi Yoshimura, Ryoko Takao-Kawabata, Hideo Matsuda, Masaru Ishii

  Apr. 2021, Nature communications, 12(1) (1), 2136 - 2136, English, International magazine

  Scientific journal


• A step towards gadolinium-free bioresponsive MRI contrast agent.

  Marzena Wyskocka-Gajda, Łukasz Przypis, Monika Olesiejuk, Tomasz Krawczyk, Anna Kuźnik, Krzysztof Nawara, Masafumi Minoshima, Fuminori Sugihara, Kazuya Kikuchi, Nikodem Kuźnik

  Feb. 2021, European journal of medicinal chemistry, 211, 113086 - 113086, English, International magazine

  Scientific journal


• High affinity modified ACE2 receptors protect from SARS-CoV-2 infection in hamsters

  Yusuke Higuchi, Tatsuya Suzuki, Takao Arimori, Nariko Ikemura, Emiko Mihara, Yuhei Kirita, Eriko Ohgitani, Osam Mazda, Daisuke Motooka, Shota Nakamura, Yusuke Sakai, Yumi Itoh, Fuminori Sugihara, Yoshiharu Matsuura, Satoaki Matoba, Toru Okamoto, Junichi Takagi, Atsushi Hoshino

  The SARS-CoV-2 spike protein binds to the human angiotensin-converting enzyme 2 (ACE2) receptor via receptor binding domain (RBD) to enter into the cell and inhibiting this interaction is a main approach to inhibit SARS-CoV-2 infection. We engineered ACE2 to enhance the affinity with directed evolution in 293T cells. Three cycles of random mutation and cell sorting achieved 100-fold higher affinity to RBD than wild-type ACE2. The extracellular domain of modified ACE2 fused to the human IgG1-Fc region had stable structure and neutralized SARS-CoV-2 without the emergence of mutational escape. Therapeutic administration protected hamsters from SARS-CoV-2 infection, decreasing lung virus titers and pathology. Engineering ACE2 decoy receptors with human cell-based directed evolution is a promising approach to develop a SARS-CoV-2 neutralizing drug that has affinity comparable to monoclonal antibodies yet displaying resistance to escape mutations of virus. Engineered ACE2 decoy receptor has a therapeutic potential against COVID-19 without viral escape mutation.

  Cold Spring Harbor Laboratory, Sep. 2020


• Bordetella Dermonecrotic Toxin Is a Neurotropic Virulence Factor That Uses CaV3.1 as the Cell Surface Receptor.

  Shihono Teruya, Yukihiro Hiramatsu, Keiji Nakamura, Aya Fukui-Miyazaki, Kentaro Tsukamoto, Noriko Shinoda, Daisuke Motooka, Shota Nakamura, Keisuke Ishigaki, Naoaki Shinzawa, Takashi Nishida, Fuminori Sugihara, Yusuke Maeda, Yasuhiko Horiguchi

  Mar. 2020, mBio, 11(2) (2), English, International magazine

  [Refereed]

  Scientific journal


• Dysregulated Expression of the Nuclear Exosome Targeting Complex Component Rbm7 in Nonhematopoietic Cells Licenses the Development of Fibrosis.

  Kiyoharu Fukushima, Takashi Satoh, Fuminori Sugihara, Yuki Sato, Toru Okamoto, Yuichi Mitsui, Sachiyo Yoshio, Songling Li, Satoshi Nojima, Daisuke Motooka, Shota Nakamura, Hiroshi Kida, Daron M Standley, Eiichi Morii, Tatsuya Kanto, Motoko Yanagita, Yoshiharu Matsuura, Takashi Nagasawa, Atsushi Kumanogoh, Shizuo Akira

  Mar. 2020, Immunity, 52(3) (3), 542 - 556, English, International magazine

  [Refereed]

  Scientific journal


• Oxygen Functional Groups on MWCNT Surface as Critical Factor Boosting T2 Relaxation Rate of Water Protons: Towards Improved CNT-Based Contrast Agents.

  Mateusz Michał Tomczyk, Sławomir Boncel, Artur Herman, Tomasz Krawczyk, Agata Jakóbik-Kolon, Mirosława Pawlyta, Maciej Krzywiecki, Artur Chrobak, Masafumi Minoshima, Fuminori Sugihara, Kazuya Kikuchi, Nikodem Kuźnik

  2020, International journal of nanomedicine, 15, 7433 - 7450, English, International magazine

  Scientific journal


• Design strategy for serine hydroxymethyltransferase probes based on retro-aldol-type reaction.

  Hiroshi Nonaka, Yuki Nakanishi, Satoshi Kuno, Tomoki Ota, Kentaro Mochidome, Yutaro Saito, Fuminori Sugihara, Yoichi Takakusagi, Ichio Aoki, Satoru Nagatoishi, Kouhei Tsumoto, Shinsuke Sando

  Feb. 2019, Nature communications, 10(1) (1), 876 - 876, English, International magazine

  [Refereed]

  Scientific journal


• Perfluorocarbon-Based 19 F MRI Nanoprobes for In Vivo Multicolor Imaging.

  Kazuki Akazawa, Fuminori Sugihara, Tatsuya Nakamura, Hisashi Matsushita, Hiroaki Mukai, Rena Akimoto, Masafumi Minoshima, Shin Mizukami, Kazuya Kikuchi

  Dec. 2018, Angewandte Chemie (International ed. in English), 57(51) (51), 16742 - 16747, English, International magazine

  [Refereed]

  Scientific journal


• Sensing caspase-1 activity using activatable 19F MRI nanoprobes with improved turn-on kinetics.

  Kazuki Akazawa, Fuminori Sugihara, Masafumi Minoshima, Shin Mizukami, Kazuya Kikuchi

  Oct. 2018, Chemical communications (Cambridge, England), 54(83) (83), 11785 - 11788, English, International magazine

  [Refereed]

  Scientific journal


• Highly Sensitive Detection of Caspase-3/7 Activity in Living Mice Using Enzyme-Responsive 19F MRI Nanoprobes.

  Kazuki Akazawa, Fuminori Sugihara, Tatsuya Nakamura, Shin Mizukami, Kazuya Kikuchi

  May 2018, Bioconjugate chemistry, 29(5) (5), 1720 - 1728, English, International magazine

  [Refereed]

  Scientific journal


• Identification of an atypical monocyte and committed progenitor involved in fibrosis.

  Takashi Satoh, Katsuhiro Nakagawa, Fuminori Sugihara, Ryusuke Kuwahara, Motooki Ashihara, Fumihiro Yamane, Yosuke Minowa, Kiyoharu Fukushima, Isao Ebina, Yoshichika Yoshioka, Atsushi Kumanogoh, Shizuo Akira

  Jan. 2017, Nature, 541(7635) (7635), 96 - 101, English, International magazine

  [Refereed]

  Scientific journal


• Modified polysaccharides as potential (19)F magnetic resonance contrast agents.

  Tomasz Krawczyk, Masafumi Minoshima, Fuminori Sugihara, Kazuya Kikuchi

  Jun. 2016, Carbohydrate research, 428, 72 - 8, English, International magazine

  [Refereed]

  Scientific journal


• Monocyte infiltration into obese and fibrilized tissues is regulated by PILRα.

  Masako Kohyama, Sumiko Matsuoka, Kyoko Shida, Fuminori Sugihara, Taiki Aoshi, Kazuki Kishida, Ken J Ishii, Hisashi Arase

  May 2016, European journal of immunology, 46(5) (5), 1214 - 23, English, International magazine

  [Refereed]

  Scientific journal


• Mesoporous silica nanoparticles for 19F magnetic resonance imaging, fluorescence imaging, and drug delivery.

  Tatsuya Nakamura, Fuminori Sugihara, Hisashi Matsushita, Yoshichika Yoshioka, Shin Mizukami, Kazuya Kikuchi

  Mar. 2015, Chemical science, 6(3) (3), 1986 - 1990, English, International magazine

  [Refereed]

  Scientific journal


• Magnetic resonance imaging of tumor with a self-traceable phosphorylcholine polymer.

  Hisatsugu Yamada, Yoshinori Hasegawa, Hirohiko Imai, Yuki Takayama, Fuminori Sugihara, Tetsuya Matsuda, Hidehito Tochio, Masahiro Shirakawa, Shinsuke Sando, Yu Kimura, Akio Toshimitsu, Yasuhiro Aoyama, Teruyuki Kondo

  Jan. 2015, Journal of the American Chemical Society, 137(2) (2), 799 - 806, English, International magazine

  [Refereed]

  Scientific journal


• Activatable 19F MRI nanoparticle probes for the detection of reducing environments.

  Tatsuya Nakamura, Hisashi Matsushita, Fuminori Sugihara, Yoshichika Yoshioka, Shin Mizukami, Kazuya Kikuchi

  Jan. 2015, Angewandte Chemie (International ed. in English), 54(3) (3), 1007 - 10, English, International magazine

  [Refereed]

  Scientific journal


• Phenylboronic acid-based (19)F MRI probe for the detection and imaging of hydrogen peroxide utilizing its large chemical-shift change.

  Hiroshi Nonaka, Qi An, Fuminori Sugihara, Tomohiro Doura, Akira Tsuchiya, Yoshichika Yoshioka, Shinsuke Sando

  2015, Analytical sciences : the international journal of the Japan Society for Analytical Chemistry, 31(4) (4), 331 - 5, English, Domestic magazine

  [Refereed]

  Scientific journal


• Multifunctional core–shell silica nanoparticles for highly sensitive (19)F magnetic resonance imaging.

  Hisashi Matsushita, Shin Mizukami, Fuminori Sugihara, Yosuke Nakanishi, Yoshichika Yoshioka, Kazuya Kikuchi

  Jan. 2014, Angewandte Chemie (International ed. in English), 53(4) (4), 1008 - 11, English, International magazine

  [Refereed]

  Scientific journal


• Gold nanoparticles coated with manganeseporphyrin that effectively shorten the longitudinal relaxation time of water molecules depending on the particle size

  Yutaka Hitomi, Kazuki Aoki, Ryosuke Miyachi, Junya Ohyama, Masahito Kodera, Tsunehiro Tanaka, Fuminori Sugihara

  2014, Chemistry Letters, 43(12) (12), 1901 - 1903, English

  [Refereed]

  Scientific journal


• An adhesive (19)F MRI chemical probe allows signal off-to-on-type molecular sensing in a biological environment.

  Tomohiro Doura, Ryunosuke Hata, Hiroshi Nonaka, Fuminori Sugihara, Yoshichika Yoshioka, Shinsuke Sando

  Dec. 2013, Chemical communications (Cambridge, England), 49(97) (97), 11421 - 3, English, International magazine

  [Refereed]

  Scientific journal


• 1SCP-05 Optically detected magnetic resonance spectroscopy of nitrogen-vacancy centers for subnanoscopic measurement in vivo(1SCP Challenges to in vivo biophysics,Symposium,The 51th Annual Meeting of the Biophysical Society of Japan)

  Igarashi Ryuji, Kumiya Yuta, Sugi Takuma, Sugihara Fuminori, Tochio Hidehito, Yoshinari Yousuke, Harada Yoshie, Shirakawa Masahiro

  The Biophysical Society of Japan General Incorporated Association, 2013, Seibutsu Butsuri, 53(1) (1), S88, English


• Real-time background-free selective imaging of fluorescent nanodiamonds in vivo.

  Ryuji Igarashi, Yohsuke Yoshinari, Hiroaki Yokota, Takuma Sugi, Fuminori Sugihara, Kazuhiro Ikeda, Hitoshi Sumiya, Shigenori Tsuji, Ikue Mori, Hidehito Tochio, Yoshie Harada, Masahiro Shirakawa

  Nov. 2012, Nano letters, 12(11) (11), 5726 - 32, English, International magazine

  [Refereed]

  Scientific journal


• Design of chemical shift switching F-19 MRI probe for imaging of reactive oxygen species

  Qi An, Tomohiro Doura, Fuminori Sugihara, Shinsuke Sando

  Aug. 2012, ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 244, English

  [Refereed]


• (19)F MRI monitoring of gene expression in living cells through cell-surface β-lactamase activity.

  Hisashi Matsushita, Shin Mizukami, Yuki Mori, Fuminori Sugihara, Masashiro Shirakawa, Yoshichika Yoshioka, Kazuya Kikuchi

  Jul. 2012, Chembiochem : a European journal of chemical biology, 13(11) (11), 1579 - 83, English, International magazine

  [Refereed]

  Scientific journal


• 3PS042 ^<19>F MRI monitoring of Gene Expression in living cells(The 50th Annual Meeting of the Biophysical Society of Japan)

  Matsushita Hisashi, Mizukami Shin, Mori Yuki, Sugihara Fuminori, Shirakawa Masahiro, Yoshioka Yoshichika, Kikuchi Kazuya

  The Biophysical Society of Japan General Incorporated Association, 2012, Seibutsu Butsuri, 52, S153, English


• Design of chemical shift-switching 19F magnetic resonance imaging probe for specific detection of human monoamine oxidase A.

  Koya Yamaguchi, Ryosuke Ueki, Hiroshi Nonaka, Fuminori Sugihara, Tetsuya Matsuda, Shinsuke Sando

  Sep. 2011, Journal of the American Chemical Society, 133(36) (36), 14208 - 11, English, International magazine

  [Refereed]

  Scientific journal


• ¹⁹F MRI detection of β-galactosidase activity for imaging of gene expression

  Shin Mizukami, Hisashi Matsushita, Rika Takikawa, Fuminori Sugihara, Masahiro Shirakawa, Kazuya Kikuchi

  Jun. 2011, Chemical Science, 2(6) (6), 1151 - 1155, English

  [Refereed]

  Scientific journal


• P-Aminophenyl alkyl ether-based ¹⁹F MRI probe for specific detection and imaging of hypochlorite ion

  Tomohiro Doura, Qi An, Fuminori Sugihara, Tetsuya Matsuda, Shinsuke Sando

  2011, Chemistry Letters, 40(12) (12), 1357 - 1359, English

  [Refereed]

  Scientific journal


• Highly redundant function of multiple AT-rich sequences as core promoter elements in the TATA-less RPS5 promoter of Saccharomyces cerevisiae.

  Fuminori Sugihara, Koji Kasahara, Tetsuro Kokubo

  Jan. 2011, Nucleic acids research, 39(1) (1), 59 - 75, English, International magazine

  [Refereed]

  Scientific journal


• The analysis of the change of rat brain from infant to adult on diffusion tensor images

  Hisanori Wakamatsu, Mika Yokoi, Yoshie Imaizumi, Kazuhiko Nakadate, Fuminori Sugihara, Takashi Ogino, Yoshiteru Seo

  2010, JOURNAL OF PHYSIOLOGICAL SCIENCES, 60, S112 - S112, English

  [Refereed]


• THE ANALYSIS OF THE SHAKING INJURY FOR INFANT RAT BRAIN BY MR TENSOR IMAGING

  Hisanori Wakamatsu, Mika Yokoi, Yoshie Imaizumi, Fuminori Sugihara, Takashi Ogino, Kazuhiko Nakadate, Yoshiteru Seo

  2009, JOURNAL OF PHYSIOLOGICAL SCIENCES, 59, 338 - 338, English

  [Refereed]


• Dual-function probe to detect protease activity for fluorescence measurement and 19F MRI.

  Shin Mizukami, Rika Takikawa, Fuminori Sugihara, Masahiro Shirakawa, Kazuya Kikuchi

  2009, Angewandte Chemie (International ed. in English), 48(20) (20), 3641 - 3, English, International magazine

  [Refereed]

  Scientific journal


• Paramagnetic relaxation-based 19f MRI probe to detect protease activity.

  Shin Mizukami, Rika Takikawa, Fuminori Sugihara, Yuichiro Hori, Hidehito Tochio, Markus Wälchli, Masahiro Shirakawa, Kazuya Kikuchi

  Jan. 2008, Journal of the American Chemical Society, 130(3) (3), 794 - 5, English, International magazine

  [Refereed]

  Scientific journal


• Paramagnetic relaxation-based F-19 MRI probe to detect protease activity

  Rika Takikawa, Shin Mizukami, Fuminori Sugihara, Masahiro Shirakawa, Kazuya Kikuchi

  2008, YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, 128, 45 - 45, English

  [Refereed]


• Magnetic resonance-based visualization of gene expression in mammalian cells using a bacterial polyphosphate kinase reporter gene.

  Sewon Ki, Fuminori Sugihara, Koji Kasahara, Hidehito Tochio, Masahiro Shirakawa, Tetsuro Kokubo

  Feb. 2007, BioTechniques, 42(2) (2), 209 - 15, English, International magazine

  [Refereed]

  Scientific journal


• A novel magnetic resonance-based method to measure gene expression in living cells.

  Sewon Ki, Fuminori Sugihara, Koji Kasahara, Hidehito Tochio, Azusa Okada-Marubayashi, Setsuko Tomita, Masahito Morita, Mitsunori Ikeguchi, Masahiro Shirakawa, Tetsuro Kokubo

  Apr. 2006, Nucleic acids research, 34(6) (6), e51, English, International magazine

  [Refereed]

  Scientific journal


• S1f2-3 In vitro and in cell NMR studies of modifier proteins(S1-f2: "Functions and dynamics of protein systems in various aspect",Symposia,Abstract,Meeting Program of EABS & BSJ 2006)

  Shirakawa Masahiro, Sakai Tomomi, Tochio Hidehito, Kokubo Tetsuro, Ki Sewon, Sugihara Fuminori, Baba Daichi, Hiroaki Hidekazu

  The Biophysical Society of Japan General Incorporated Association, 2006, Seibutsu Butsuri, 46(2) (2), S119, English

• RNA解析を用いたCOVID-19によるARDSの病態解明

  伊藤 弘, 石川 昌和, 吉村 旬平, 劉 祐誠, 松本 寿健, 杉原 文徳, 榊原 修平, 奥崎 大介, 平田 陽彦, 小倉 裕司, 織田 順

  (一社)日本集中治療医学会, Jun. 2023, 日本集中治療医学会雑誌, 30(Suppl.1) (Suppl.1), S586 - S586, Japanese


• Development of elastic perfluorocarbon-encapsulated polymer nanoparticles for ¹⁹F MRI contrast agents

  小西祐輝, 杉原文徳, 杉原文徳, 蓑島維文, 蓑島維文, 藤原耕平, 内橋貴之, 菊地和也, 菊地和也

  2023, 日本化学会春季年会講演予稿集(Web), 103rd


• 血漿タンパク質量分析による分子病態分類は重症熱傷の予後と関連する

  大西伸也, 松本寿健, 杉原文徳, 大須賀章倫, 小倉裕司, 織田順

  2023, 日本熱傷学会総会・学術集会プログラム・抄録集, 49th


• Immunomolecular pathogenesis of COVID-19-induced ARDS comparing lung and systemic immune responses

  光山裕美, 松本寿健, 杉原文徳, 梅村穣, 藤見聡, 藤見聡, 小倉裕司

  (一社)日本集中治療医学会, 2022, 日本集中治療医学会学術集会(Web), 49th(Suppl.1) (Suppl.1), 313 - 313, Japanese


• Pathogenesis of severe COVID-19 by whole blood mRNA/miRNA network analysis

  戸上由貴, 松本寿健, 松原庸博, 蛯原健, 石川昌和, 杉原文徳, 瀬尾茂人, 奥崎大介, 平田陽彦, 小倉裕司

  2022, 日本集中治療医学会学術集会(Web), 49th


• PEA法を用いた血漿プロテオミクス解析による重症新型コロナ感染症の分子病態型と患者層別化マーカー開発

  松本寿健, 蛯原健, 松原庸博, 戸上由貴, 舘野丈太郎, 松浦裕司, 松浦裕司, 児島嵩, 平田陽彦, 杉原文徳, 瀬尾茂人, 奥崎大介, 小倉裕司

  2021, 日本人類遺伝学会大会プログラム・抄録集, 66th (CD-ROM)


• パーフルオロカーボン内包シリカナノ粒子を用いたカテプシンK活性の高感度かつ選択的¹⁹F MRイメージング

  奥西敦也, 赤澤一樹, 杉原文徳, 水上進, 菊地和也, 菊地和也

  10 May 2018, JSMI Report, 11(2) (2), 92, Japanese


• パーフルオロカーボン内包シリカナノ粒子を用いたカテプシンK活性の高感度かつ選択的¹⁹F MRイメージング

  奥西敦也, 赤澤一樹, 杉原文徳, 水上進, 菊地和也, 菊地和也

  06 Mar. 2018, 日本化学会春季年会講演予稿集(CD-ROM), 98th, ROMBUNNO.1D3‐34, Japanese


• パーフルオロカーボン内包シリカナノ粒子を用いた多機能性¹⁹F MRI造影剤の開発

  穐本怜奈, 赤澤一樹, 杉原文徳, 菊地和也, 菊地和也

  18 Aug. 2017, バイオイメージング, 26(2) (2), 102, Japanese


• パーフルオロカーボン内包シリカナノ粒子を用いたシグナル増大型¹⁹F MRIナノプローブの開発

  赤澤一樹, 穐本怜奈, 杉原文徳, 水上進, 菊地和也, 菊地和也

  03 Mar. 2017, 日本化学会春季年会講演予稿集(CD-ROM), 97th, ROMBUNNO.2PB‐204, Japanese


• 多重共鳴MRI画像法による自然発症2型糖尿病モデルの脂質成分の検討

  上田裕紀, 野嶋孝次, 杉原文徳

  2017, 日本老年医学会雑誌, 54(4) (4), 621(J‐STAGE), Japanese


• Neuronal activities concerning the conditioned taste aversion with immune reactions

  Chizuko Inui-Yamamoto, Fuminori Sugihara, Ting Chen, Yoshichika Yoshioka, Satoshi Wakisaka

  Nov. 2016, CHEMICAL SENSES, 41(9) (9), E173 - E174, English

  Summary international conference


• パーフルオロカーボン内包シリカナノ粒子を用いたシグナル増大型¹⁹F MRIナノプローブの開発

  赤澤一樹, 杉原文徳, 吉岡芳親, 水上進, 水上進, 菊地和也, 菊地和也

  25 Apr. 2016, JSMI Report, 9(2) (2), 116, Japanese


• 生体における免疫応答を可視化するカスパーゼ‐1活性検出¹⁹F MRI造影剤の開発

  赤澤一樹, 杉原文徳, 吉岡芳親, 水上進, 水上進, 菊地和也, 菊地和也

  10 Mar. 2016, 日本化学会春季年会講演予稿集(CD-ROM), 96th, ROMBUNNO.4A4‐32, Japanese


• 高感度¹⁹F MRI造影剤を用いたマルチスペクトルイメージング

  有薗賢志, 杉原文徳, 赤澤一樹, 吉岡芳親, 向井大陽, 水上進, 菊地和也

  10 Mar. 2016, 日本化学会春季年会講演予稿集(CD-ROM), 96th, ROMBUNNO.4A4‐31, Japanese


• マルチモーダルイメージング可能な薬物送達メソポーラスシリカナノ粒子

  石田健一郎, 杉原文徳, 松下尚嗣, 中村竜也, 吉岡芳親, 水上進, 水上進, 菊地和也, 菊地和也

  10 Mar. 2016, 日本化学会春季年会講演予稿集(CD-ROM), 96th, ROMBUNNO.4A4‐33, Japanese


• 常磁性緩和促進効果に基づいた刺激応答性¹⁹F MRI造影剤の開発

  赤澤一樹, 中村竜也, 杉原文徳, 吉岡芳親, 水上進, 菊地和也

  27 Apr. 2015, JSMI Rep, 8(2) (2), 133, Japanese


• 分子標的MRイメージングのための新手法

  長谷川 嘉則, 山田 久嗣, 木村 祐, 今井 宏彦, 高山 裕生, 杉原 文徳, 松田 哲也, 杤尾 豪人, 白川 昌宏, 山東 信介, 青山 安宏, 年光 昭夫, 近藤 輝幸

  May 2013, 日本分子イメージング学会第8回学術集会, MS-11(P-46)

  Summary national conference


• 安定同位元素ラベル化生体適合性高分子プローブの開発と磁気共鳴イメージングへの応用

  長谷川 嘉則, 山田 久嗣, 木村 祐, 杤尾 豪人, 白川 昌宏, 杉原 文徳, 今井 宏彦, 高山 裕生, 松田 哲也, 矢野 哲哉, 山東 信介, 青山 安宏, 年光 昭夫, 近藤 輝幸

  Mar. 2013, 日本化学会第93回春季年会, 1E3-29

  Summary national conference


• 高感度多重共鳴NMR解析のための安定同位元素ラベル化生体適合性高分子プローブ

  長谷川 嘉則, 山田 久嗣, 木村 祐, 杤尾 豪人, 白川 昌宏, 杉原 文徳, 今井 宏彦, 高山 裕生, 松田 哲也, 矢野 哲哉, 山東 信介, 青山 安宏, 年光 昭夫, 近藤 輝幸

  Mar. 2013, 日本化学会第93回春季年会, 1E-3-28

  Summary national conference


• 安定同位体元素を集積化した生体適合性高分子プローブの合成と多重共鳴NMR法による機能評価

  長谷川 嘉則, 山田 久嗣, 木村 祐, 杤尾 豪人, 白川 昌宏, 杉原 文徳, 今井 宏彦, 高山 裕生, 松田 哲也, 矢野 哲哉, 山東 信介, 青山 安宏, 年光 昭夫, 近藤 輝幸

  Oct. 2012, 第2回CSJ化学フェスタ, P5-79

  Summary national conference


• Rice retrogradation slowing down by alpha-glucosidase

  Yamaguchi Hideyuki, Okamoto Takeshi, Sugihara Buntoku, Shimba Nobuhisa, Wakabayashi Hidehiko, Shirakawa Masahiro, Suzuki Ei-ichiro

  <BR> 【背景】<BR> 国内の中食市場を中心に様々な米飯食品が販売されているが、炊き立ての食感を長く維持することは難しい。食感低下の原因は、炊飯により澱粉結晶が解け、柔らかく粘りのある糊化状態となったものが、澱粉の再結晶化により硬く粘りの無い状態になることと考えられている。我々は、糖鎖を基質とするα-グルコシダーゼを米に作用させることにより、炊飯後の経時的な食感低下が抑制されることを見出した。そして、NMRイメージングによって視覚的にα-グルコシダーゼの効果を観察することに成功した。<BR>【方法】<BR> 米の水浸漬時にα-グルコシダーゼを加え、室温で24時間反応させた後、炊飯を行った。10mmφのNMR試料管に、α-グルコシダーゼ処理、無処理の炊飯米を並べて封入し、炊飯直後から24時間後までのNMR緩和時間T2の経時変化を追跡した。この結果について、テクスチャーアナライザーによる硬さと粘り、及びアミラーゼ・プルラナーゼ法による糊化度の経時変化と比較した。<BR>【結果・考察】<BR> α-グルコシダーゼ処理した炊飯米は、T2の経時変化が少ないことが判った。T2は分子の運動性に対応して変化する量であり、α-グルコシダーゼ処理することによって炊飯米中の水分子の運動性が一定に保たれる傾向があると考えられる。これは、粘りと硬さ、及び糊化度の経時変化と同様な傾向であり、T2は米飯老化の様子を観察する良いパラメーターであることが判った。

  The Japan Society of Cookery Science, 2008, Abstracts of the Annual Meeting of the Japan Society of Cookery Science, 20(0) (0), 114 - 114, Japanese


• The analysis of the shaking injury for hippocampus areas of infant rat brain by diffusion weighted imaging

  Hisanori Wakamatsu, Yokoi Mika, Imaizumi Yoshie, Nakadate Kazuhiko, Sugihara Fuminori, Ogino Takashi, Seo Yoshiteru

  We participate in the project to establish the technique to detect an injury by shaking for brain in childhood resulted in a nerve disease at the early stage. The aim of our research as the development of method of detecting the injury with the diffusion weighted image by MRI using laboratory animals, evaluating the level of injury by using Fractional anisotropy (FA) and Trace index (TI) obtained from those images. In this presentation, we show the fluctuation of FA and TI values in the image of hippocampus of the brain of infant rat. Diffusion weighted images of hippocampus areas of postnatal 4th to 13th day rat were acquired. The FA value at molecular cell layer (Mol) is higher than that at pyramidal cell layer (Py) and dentate gyrus (DG), and the TI value at DG is lower than others. These results were similar to previous data of adult mouse though a significant conclusion was not obtained. However variations of FA value at Mol and Py that made a peak on the 10th day, and both were significantly different from at DG (two-way ANOVA: DG-Mol P=0.036, DG-Py P=0.028). But TI value at all areas had not show a peak in each day age. These results possibly are related to the expansion of the nerve fiber and the restructuring of the synapse that occur in the infant age rat brain. <b>[J Physiol Sci. 2008;58 Suppl:S151]</b>

  PHYSIOLOGICAL SOCIETY OF JAPAN, 2008, Proceedings of Annual Meeting of the Physiological Society of Japan, 2008(0) (0), 151 - 151


• テクノ・トレンド 磁気共鳴法を利用した生細胞での遺伝子発現解析

  古久保 哲朗, 奇 世媛, 杉原 文徳

  羊土社, Sep. 2006, バイオテクノロジージャーナル, 6(5) (5), 605 - 608, Japanese


• MRIで見る米飯の老化

  山口 秀幸, 岡本 武, 杉原 文徳, 若林 秀彦, 白川 昌宏, 鈴木 榮一郎

  01 Mar. 2006, バイオサイエンスとインダストリー = Bioscience & industry, 64(3) (3), 143 - 144, Japanese


• High-field mouse MRI probe for stereotaxic analysis

  Wakamatsu Hisanori, Yokoi Mika, Imaizumi Yoshie, Sugihara Fuminori, Ogino Takashi, Seo Yoshiteru

  Our new project is to detect minimal brain injury in the shaken-baby syndrome. We will take a rat model that was established by Ueda et al (Neuroscience Letters, <B>385</B> 82-86 2005). A high-spatial resolution image and a diffusion analysis can be useful to detect and follow pathological changes in the brain. It is reasonable to take a higher field to get a better image of small brains of neonatal, infantile, and juvenile rats that are similar size to mouse. In order to establish a basic system in vertical high-field MR micro-imaging system, we had made a probe with the stereotaxic coordinates for mice inside a gradient system. A set of the conventional fixation devices (a bite bar and a pair of ear bars) was installed in an acrylic tube. The position of the bite bar can be adjusted to the level position. 1% enflurane in a gas mixture of 36% O<SUB>2</SUB>-2%CO2-62% NO delivered through the tracheal canula by artificial ventilator. The body temperature was kept in adequate range by a hot water circuit, and the temperature was monitored by fluorescence thermometer. ECG was also monitored by PowerLab. Using this probe, we can get reasonable quality of gradient-echo image, spin-echo diffusion image, and multi-slice multi-spin-echo image in 100 &mu;m pixel resolution with a slice thickness of 0.5 mm. Vital conditions of mice kept constant for 4 hours and longer. We are now trying to detect a minimal hemorrhage in the infant rat brain that was shaken by a shaking machine. <b>[J Physiol Sci. 2006;56 Suppl:S213]</b>

  PHYSIOLOGICAL SOCIETY OF JAPAN, 2006, Proceedings of Annual Meeting of the Physiological Society of Japan, 2006(0) (0), 213 - 213


• In Vivo Molecular Imaging by Using MRI

  SUGIHARA Fuminori, TOCHIO Hidehito

  MRI(magnetic resonance imaging) is a non-destructive and non-invasive visualizing method, and has been widely used in clinical medicine and biological studies. Combining MRI with recent developments of molecular and cellular biology together with genetically modified animals is now opening up a new era of MRI, where various biological information, rather than only morphological shape, can be obtained in vivo at cellular and sub-cellular level by sophisticatedly designed MR reporter systems. This short review starts with a brief description of the basic concept of MRI and introduces recent topics.

  The Biophysical Society of Japan General Incorporated Association, 25 Nov. 2004, Biophysics, 44(6) (6), 271 - 275, Japanese

• 線維症における病原性マクロファージの動態と機能解析

  杉原 文徳

  日本学術振興会, 科学研究費助成事業, 研究活動スタート支援, 神戸大学, 31 Jul. 2024 - 31 Mar. 2026


• 網羅的生体分子情報に基づく重症熱傷病態の中心的分子解明と新規治療薬開発

  大西 伸也, 奥崎 大介, 杉原 文徳, 清水 健太郎, 大須賀 章倫, 小倉 裕司, 松本 寿健

  日本学術振興会, 科学研究費助成事業, 基盤研究(C), 大阪大学, 01 Apr. 2022 - 31 Mar. 2025

  網羅的生体分子情報とは、メッセンジャーRNAやノンコーディングRNAといった遺伝子発現や遺伝子産物であるタンパクなどの変化に関する情報である。この大量の情報を統合して解析するバイオインフォマティクス技術が発展しており、あらゆる疾患の新たな病態解明につながっている。本研究では、熱傷における遺伝子やタンパクの変化を網羅的に測定し、統合解析を行うとともに、明らかとなった中心的分子の役割を基礎実験にて解明することが目標である。以下の2つに焦点を当てている。①臨床検体を用いて重症熱傷において中心的役割を担う分子(遺伝子/タンパク)を同定し治療効果/重症度/予後との関連性を評価する。②同定された中心的分子に関して熱傷マウスモデルを用いて役割を解明する。 初年度は、1：臨床検体の収集、2：重症熱傷の臨床検体を用いた網羅的タンパク発現の解析を行った。①に関してはより臨床表現型を反映する網羅的タンパク解析を優先して行っている。既存検体を用いた血漿中に含まれるタンパクの質量分析の測定結果について、詳細な臨床情報を組み合わせ、より多角的に解析を行った。健常者との比較において23個のタンパクの変化が有意であり、酸素代謝やコレステロールのエステル化等のプロセスに関わっていることが分かった。転帰の違いによる比較においては、10個のタンパクの変化が有意であることが分かり、その中でもヘモグロビンサブユニットやTTR、SERPINF2といったタンパクが中心的分子であることが分かった。


• Immunomolecular pathogenesis of systemic and local lung immunity in ARDS based on integrated analysis

  光山 裕美, 松本 寿健, ウィング ジェイムス, 奥崎 大介, 杉原 文徳, 小倉 裕司, 藤見 聡, 梅村 穣

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (C), Osaka University, 01 Apr. 2022 - 31 Mar. 2025

  急性呼吸窮迫症候群(ARDS) における肺局所免疫と全身免疫の双方の観点から、免疫細胞間およびサイトカイン相互の複雑な関連を明らかにすることが目的っである。当該年度では、 ARDS患者の血液と気管支肺胞洗浄液（BALF）の網羅的生体情報を収集・測定し、免疫系ネットワークの全体像を把握することを主眼とした。BALFは36サンプル収集し、対応する末梢血のPBMCおよび血漿を同時に採取した。BALF中の細胞およびPBMCはCyTOFを用いて測定した。BALFの上澄、血漿は質量分析を用いてタンパクを網羅的に測定した。これらのデータの予備解析を行なった。質量分析でのタンパク解析では肺局所では炎症の活性化がみられるものの、末梢血においては炎症が抑制されている結果であった。単一細胞解析では、末梢血とBALF内においてCXCR4＋組織常在型T細胞が増加しており、活性化した末梢性のT細胞がCXCR4を介して、肺局所に動員され、肺障害に寄与している可能性が示唆された。また、末梢血では骨髄由来抑制細胞（MDSC）が増加し、抗原提示機能が低下したマクロファージ細胞が増加していたが、BALFでは末梢血の単球由来の炎症性肺胞マクロファージが増加していた。 次年度にこれらのデータを、単一細胞解析においてさらに詳細に解析し、質量分析により得られたデータを詳細に解析することで、ARDS患者の局所と全身における免疫系ネットワークの評価を行う予定である。


• 脂肪細胞による全身性炎症反応制御：オートファジーとアディポカイン産生

  蛯原 健, 福田 士郎, 喜多 俊文, 杉原 文徳, 清水 健太郎, 小倉 裕司, 松本 寿健

  日本学術振興会, 科学研究費助成事業, 基盤研究(C), 大阪大学, 01 Apr. 2022 - 31 Mar. 2025

  本研究では以下の2つに注目し研究を行う。1)SIRS患者（敗血症、熱傷、CVID-19）の脂肪組織におけるオートファジーの役割を明らかにする。2）SIRS患者におけるアディポネクチン/Tカドヘリンの役割を明らかにする。 本年度は2）に対して重症熱傷患者を対象にアディポネクチン、T-カドヘリンの動態を明らかにした。対象は重症熱傷64例、健常コントロール16例である。熱傷患者のうち12例は死亡した。血漿中の、アディポネクチンと3種類（130kDa、100kDa、30kDa)のT-カドヘリンを複数回、ELISA法を用いて測定した。また併せて血管内皮障害のマーカーとしてPAI-1、Hyaluronan、Syndecan-1、Glypican-1もELISA法を用いて測定した。結果として、死亡例では来院時に採取した血漿のアディポネクチンが有意に高値を示した。一方、T-カドヘリンと死亡との関連は認めなかった。熱傷では初期に血管内皮障害を契機とした血管透過性の亢進が生じるが、循環を維持するために大量の輸液投与をおこなう。この初期24時間の輸液量と来院時に採取した血漿中のTカドヘリン（100kDa)が逆相関を示していた。このT-カドヘリンは血管内皮障害が生じた場合上昇する、PAI-1、Syndecan-1、Hyaluronanとも逆相関を認めたことから、血管内皮障害の程度に応じて血中のTカドヘリンが低下していることが予想された。アディポネクチンはTカドヘリンに結合し細胞保護的に働くことがわかっており、熱傷侵襲により傷ついた血管内皮の修復にTカドヘリンが使用されている可能性が考えられた。


• Development of in vivo oxygen environment analysis by using MRI

  杉原 文徳

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Osaka University, 01 Apr. 2017 - 31 Mar. 2019

  本研究では酸素によりMRIのコントラストが強調される酸素増感MRI(OE-MRI)法を用い、ガンや炎症などで見られるOE-MRIのコントラストがどのような因子と関連するかを他の解析方法と比較することで明らかとし、生体中での酸素動態を可視化するツールとして用いることがきないか検討した。 測定手法の検討では酸素投与によるコントラストの増加に加え、呼気酸素濃度を下げることでコントラストの低下を引き起こすことを見出した。この酸素濃度の異なる画像の差分から血管造影にによる比較的太い血管以外にも微細な血管分布を示すような画像が得られることが分かった。13C標識グルコースがラクトースへ代謝される過程を生体中でモニターできる代謝MRI方法とOE-MRIを組み合わせることで、in vivoでの相関性をガン組織中で測定し、その組織部位ごとでのコントラストの相関性のある因子を同定することを試みた。性質の異なる細胞の皮下腫瘍を形成させMRI撮像し、血管走行を観察するために蛍光デキストランを用いた組織切片を作成していった。しかし、蛍光により血管分布をMRIで観察した視野と比較するには想定以上に時間がかかり、3次元的な分布を再構築することは困難であると判断した。代謝MRIとOE-MRIの相関性については細胞種によっても異なるのみでなく、腫瘍内部の状況によってもばらつきがあり、解析例数を増やした検証が必要と考えられた。 脳血梗塞による炎症モデルを用いた評価では、急性期には酸素の造影効果がみられたが、時間経過によりその効果は見られなくなることを見出した。一方、自己抗体産生による慢性的な炎症では、ガドリニウム造影による血管関門の破綻は観察されるが、酸素造影では同様の効果は得られなかった。これらの結果から、急性的な炎症状態であるかどうかを評価する方法の一つとしてOE-MRIを用いることが可能であると考えられた。


• Design, Synthesis, and Biological Application of in vivo Imaging Probes

  Kikuchi Kazuya, HORI Yuichiro, SUGIHARA Fuminori, MINOSHIMA Masafumi

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (S), Osaka University, 31 May 2013 - 31 Mar. 2018

  In this study, we designed and synthesized functional chemical probes for visualization of physiological phenomena in living animals. (1) We developed highly sensitive 19F MRI nanoprobes composed of perfluorocarbon-encapsulated silica nanoparticles. We used the nanoprobes for visualization of tumor tissue, detection of enzyme activity, and multicolor imaging. (2) We developed pH-responsive fluorescent probes for imaging acidic pH regions in bone tissue during osteoclastic bone resorption. The real-time images revealed actual behaviors of activated osteoclasts in living mice. These functional molecular probes provide new tools for analyzing dynamics and function of biomolecules in vivo.


• The analysis of relationships between the changes in taste preference and immune function

  Inui Chizuko, YOSHIOKA Yoshichika, SUGIHARA Fuminori, MORI Yuki, CHIN Tei

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Challenging Exploratory Research, Osaka University, 01 Apr. 2015 - 31 Mar. 2017

  Some reports show that rodents can acquire aversion to the taste stimulus paired with LPS, which induces inflammatory reaction. However, the brain mechanisms in the conditioned taste aversion (CTA) with immune reactions remain obscure. Therefore, to elucidate the brain mechanism in the retrieval of CTA with LPS, we tried to detect the brain activities using a manganese-enhanced MRI. C57BL/6 male mice were conditioned by i.p. injection of saline, LiCl, or LPS, after drinking saccharin solution. On the test day, all mice were given saccharin solution and scanned by 11.7 T MRI. Our results showed that the signal intensity of the amygdala in the LPS- and LiCl-groups were higher than that in the Saline-group. The signal intensity of the dorsomedial hypothalamus (DMH) only in the LPS-group was higher. The DMH has a role of the body temperature control. Therefore, there is a possibility that the DMH might have an important role in the relationship between taste preference and immune system.


• Development and application of multiple quantum coherence MRI for the analysis of lipid composition in vivo

  SUGIHARA Fuminori

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Young Scientists (B), Osaka University, 01 Apr. 2012 - 31 Mar. 2014

  Adipose tissue is known as energy storage and also heat generator at the brown adipose tissue. Excess amount of energy intake such as sugar and fat lead obesity, in which the adipose tissue hold extra lipid and also secret inflammation cytokine. Imaging devices are used for diagnosis of obesity. MRI is one of the imaging tool and able to observe not only the area of adipose tissue but also used for analysis of lipid composition in detail. In this study, multiple quantum coherence MRI method was used for imaging lipid composition in vivo using model animal under various conditions. As the results of time course observation with induced obesity, it was revealed that the lipid composition of saturated-unsaturated lipid was change at the adipose tissue.

• 重合体、前記重合体を用いた核磁気共鳴分析用または磁気共鳴イメージング用の造影剤、化合物、前記重合体を用いた核磁気共鳴分析方法および磁気共鳴イメージング方法

  近藤 輝幸, 山田 久嗣, 長谷川 嘉則, 木村 祐, 青山 安宏, 杤尾 豪人, 白川 昌宏, 杉原 文徳, 松田 哲也, 山東 信介, 南 昌人, 山内 文生, 矢野 哲哉, 都築 英寿

  特願2013-103159, 15 May 2013, キヤノン株式会社, 特開2014-001371, 09 Jan. 2014, 特許第6234064号, 02 Nov. 2017

  Patent right

  IRI


• 重合体、前記重合体を用いた核磁気共鳴分析用または磁気共鳴イメージング用の造影剤、化合物、前記重合体を用いた核磁気共鳴分析方法および磁気共鳴イメージング方法

  近藤 輝幸, 山田 久嗣, 長谷川 嘉則, 木村 祐, 青山 安宏, 杤尾 豪人, 白川 昌宏, 杉原 文徳, 松田 哲也, 山東 信介, 南 昌人, 山内 文生, 矢野 哲哉, 都築 英寿

  特願2013-103159, 15 May 2013, 国立大学法人京都大学, キヤノン株式会社, 特開2014-001371, 09 Jan. 2014

  Patent right

  IRI


• 非破壊的な遺伝子発現モニタリング技術を活用したハイスループット型薬物スクリーニング手法

  古久保 哲朗, 白川 昌宏, 杉原 文徳

  特願2005-064852, 09 Mar. 2005, 横浜市, 特開2006-246736, 21 Sep. 2006

  Patent right

  IRI