Elucidation of the pathogenesis of endoplasmic reticulum stress in congenital neutropenia based on proteome analysis

• Project/Area Number: 21K15905
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 52050:Embryonic medicine and pediatrics-related
• Research Institution: Hiroshima University
• Principal Investigator: Mizoguchi Yoko 広島大学, 医系科学研究科(医), 助教 (30750533)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Project Status: Completed (Fiscal Year 2023)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2022: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2021: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: 先天性好中球減少症 / iPSC / UPR / 酸化ストレス / プロテオーム / 小胞体ストレス / 好中球減少症 / ELANE異常症

Outline of Research at the Start

先天性好中球減少症(CN)は、好中球減少による細菌感染症の反復を特徴とする遺伝性疾患であり、病態として小胞体ストレス反応（UPR）の関与が指摘されている。研究代表者は過去にCNで頻度の高い責任遺伝子であるELANE及びSRP54遺伝子変異を有するCN患者好中球において、網羅的プロテオーム解析を行った。その中でCNの共通病態であるUPRに最も深く関係していると考えられる分子を抽出した。本研究ではそれらの分子の発現低下がCNの共通病態（UPR、好中球分化障害、細胞死）の上流にあるとの病態仮説をたて、骨髄CD34陽性細胞及びiPS細胞モデルを用いたCNの分子病態の解明を目指す。

Outline of Final Research Achievements

Based on the hypothesis that decreased GSTP1 expression might be involved in the pathogenesis of CN, gene disruption of GSTP1 was performed using the CRISPR/Cas9 method in healthy-derived iPS cells. However, no differentiation defect was found. This indicates that a single deletion of GSTP1 alone does not affect neutrophil differentiation. For further analysis, we disrupted the ELANE gene using the CRISPR/Cas9 method in patient-derived iPS cells (CN-iPSC-NE KO) and examined whether the impairment of neutrophil differentiation was improved. CN-iPSC-NE KO showed an improved neutrophil differentiation defect. RNA-seq of CN-iPSC and CN-iPSC-NE KO-derived neutrophil progenitors revealed dysregulation of the antioxidant pathway in CN-iPSC. This suggests that not only UPR but also oxidative stress is involved in the pathogenesis of CN.

Academic Significance and Societal Importance of the Research Achievements

CNの病態にUPRのみならず酸化ストレスも関与している可能性が示唆され、薬剤スクリーニングを通して好中球分化障害を改善する分子を同定できれば、患者のQOLを改善する可能性がある。

Research Products

• [Journal Article] Generation of complex bone marrow organoids from human induced pluripotent stem cells (2024)
  • Author(s): Frenz-Wiessner S, Fairley SD, Buser M, Goek I, Salewskij K, Jonsson G, Illig D, Zu Putlitz B, Petersheim D, Li Y, Chen PH, Kalauz M, Conca R, Sterr M, Geuder J, Mizoguchi Y, Megens RTA, Linder MI, Kotlarz D, Rudelius M, Penninger JM, Marr C, Klein C.
  • Journal Title: Nature Methods Volume: - Issue: 5 Pages: 868-881
  • DOI: 10.1038/s41592-024-02172-2
• [Journal Article] Human genetic defects in SRP19 and SRPRA cause severe congenital neutropenia with distinctive proteome changes (2023)
  • Author(s): Linder Monika I.、Mizoguchi Yoko、Hesse Sebastian、et al.
  • Journal Title: Blood Volume: 141 Issue: 6 Pages: 645-658
  • DOI: 10.1182/blood.2022016783
• [Journal Article] A complementary approach for genetic diagnosis of inborn errors of immunity using proteogenomic analysis (2023)
  • Author(s): Sakura F, Noma K, Asano T, Tanita K, Toyofuku E, Kato K, Tsumura M, Nihira H, Izawa K, Mitsui-Sekinaka K, Konno R, Kawashima Y, Mizoguchi Y, Karakawa S, Hayakawa S, Kawaguchi H, Imai K, Nonoyama S, Yasumi T, Ohnishi H, Kanegane H, Ohara O, Okada S
  • Journal Title: PNAS Nexus Volume: 2 Issue: 4 Pages: 1-12
  • DOI: 10.1093/pnasnexus/pgad104
  • Peer Reviewed / Open Access
• [Journal Article] Structural basis of spike RBM-specific human antibodies counteracting broad SARS-CoV-2 variants (2023)
  • Author(s): Kiyomi Shitaoka, Akifumi Higashiura, Yohei Kawano, Akima Yamamoto, Yoko Mizoguchi, Takao Hashiguchi , Norihisa Nishimichi, Shiyu Huang, Ayano Ito, Shun Ohki, Miyuki Kanda, Tomohiro Taniguchi, Rin Yoshizato, Hitoshi Azuma, Yasuo Kitajima, Yasuyuki Yokosaki, Satoshi Okada, Takemasa Sakaguchi, Tomoharu Yasuda
  • Journal Title: Communications Biology Volume: 6 Issue: 1 Pages: 1-14
  • DOI: 10.1038/s42003-023-04782-6
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Analyzing mitochondrial respiration of human induced pluripotent stem cell-derived myeloid progenitors using Seahorse technology (2023)
  • Author(s): Fan Yanxin、Mizoguchi Yoko、Tatematsu Megumi、Linder Monika I.、Frenz Stephanie、Choi Jongsu、Klein Christoph
  • Journal Title: STAR Protocols Volume: 4 Issue: 1 Pages: 102073-102073
  • DOI: 10.1016/j.xpro.2023.102073
• [Journal Article] Human STAT1 gain of function with chronic mucocutaneous candidiasis: A comprehensive review for strengthening the connection between bedside observations and laboratory research (2023)
  • Author(s): Asano Takaki、Noma Kosuke、Mizoguchi Yoko、Karakawa Shuhei、Okada Satoshi
  • Journal Title: Immunological Reviews Volume: 322 Issue: 1 Pages: 81-97
  • DOI: 10.1111/imr.13300
  • Peer Reviewed
• [Journal Article] Isolated Chronic Mucocutaneous Candidiasis due to a Novel Duplication Variant of IL17RC (2023)
  • Author(s): Noma Kosuke、Tsumura Miyuki、Nguyen Tina、Asano Takaki、Sakura Fumiaki、Tamaura Moe、Imanaka Yusuke、Mizoguchi Yoko、Karakawa Shuhei、Hayakawa Seiichi、Shoji Takayo、Hosokawa Junichi、Izawa Kazushi、Ling Yun、Casanova Jean-Laurent、Puel Anne、Tangye Stuart G.、Ma Cindy S.、Ohara Osamu、Okada Satoshi
  • Journal Title: Journal of Clinical Immunology Volume: 44 Issue: 1 Pages: 18-18
  • DOI: 10.1007/s10875-023-01601-9
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Age‐specific incidence of joint disease in paediatric patients with haemophilia: A single‐centre real‐world outcome based on consecutive US examination (2023)
  • Author(s): Mizoguchi Yoko、Tani Chihiro、Aizawa Mika、Tomioka Keita、Shimomura Maiko、Nishimura Shiho、Matsubara Yoshiko、Iwaki Daisuke、Tanaka Kiyoto、Kawaguchi Hiroshi、Nakashima Yuko、Mikami Yukio、Okada Satoshi、Kobayashi Masao
  • Journal Title: Haemophilia Volume: 29 Issue: 5 Pages: 1359-1365
  • DOI: 10.1111/hae.14848
• [Journal Article] Human T-bet governs the generation of a distinct subset of CD11chighCD21low B cells (2022)
  • Author(s): Yang R, Avery DT, Jackson KJL, Ogishi M, Benhsaien I, Du L, Ye X, Han J, Rosain J, Peel JN, Alyanakian MA, Neven B, Winter S, Puel A, Boisson B, Payne KJ, Wong M, Russell AJ, Mizoguchi Y, . et al.
  • Journal Title: Science Immunology Volume: 7 Issue: 73 Pages: 3277-3290
  • DOI: 10.1126/sciimmunol.abq3277
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Neutralizing Type I Interferon Autoantibodies in Japanese Patients with Severe COVID-19 (2022)
  • Author(s): Eto S, Nukui Y, Tsumura M, Nakagama Y, Kashimada K, Mizoguchi Y, Utsumi T, Taniguchi M, Sakura F, Noma K, Yoshida Y, Ohshimo S, Nagashima S, Okamoto K, Endo A, Imai K, Kanegane H, Ohnishi H, Hirata S, Sugiyama E, Shime N, Ito M, Ohge H, Kido Y, Bastard P, Casanova JL, Ohara O, Tanaka J, Morio T, Okada S
  • Journal Title: Journal of Clinical Immunology Volume: 42 Issue: 7 Pages: 1360-1370
  • DOI: 10.1007/s10875-022-01308-3
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Mendelian susceptibility to mycobacterial diseases: state-of-the-art (2022)
  • Author(s): Noma Kosuke、Mizoguchi Yoko、Tsumura Miyuki、Okada Satoshi
  • Journal Title: Clinical Microbiology and Infection Volume: － Issue: 11 Pages: 00143-4
  • DOI: 10.1016/j.cmi.2022.03.004
  • Peer Reviewed / Open Access
• [Journal Article] HAX1-dependent control of mitochondrial proteostasis governs neutrophil granulocyte differentiation (2022)
  • Author(s): Fan Yanxin、Murgia Marta、Linder Monika I.、Mizoguchi Yoko、Wang Cong、Lyszkiewicz Marcin、Zietara Natalia、Liu Yanshan、Frenz Stephanie、Sciuccati Gabriela、Partida-Gaytan Armando、Alizadeh Zahra、Rezaei Nima、Rehling Peter、Dennerlein Sven、Mann Matthias、Klein Christoph
  • Journal Title: Journal of Clinical Investigation Volume: 132 Issue: 9 Pages: 1-18
  • DOI: 10.1172/jci153153
  • Peer Reviewed
• [Journal Article] Enhanced osteoclastogenesis in patients with MSMD due to impaired response to IFN-γ (2022)
  • Author(s): Tsumura Miyuki、Miki Mizuka、Mizoguchi Yoko、Hirata Osamu、Nishimura Shiho、Tamaura Moe、Kagawa Reiko、Hayakawa Seiichi、Kobayashi Masao、Okada Satoshi
  • Journal Title: Journal of Allergy and Clinical Immunology Volume: 149 Issue: 1 Pages: 252-261
  • DOI: 10.1016/j.jaci.2021.05.018
  • Peer Reviewed / Open Access
• [Journal Article] Inherited CARD9 Deficiency in a Child with Invasive Disease Due to Exophiala dermatitidis and Two Older but Asymptomatic Siblings (2021)
  • Author(s): Imanaka Yusuke、Taniguchi Maki、Doi Takehiko、Tsumura Miyuki、Nagaoka Rie、Shimomura Maiko、Asano Takaki、Kagawa Reiko、Mizoguchi Yoko、Karakawa Shuhei、Arihiro Koji、Imai Kohsuke、Morio Tomohiro、Casanova Jean-Laurent、Puel Anne、Ohara Osamu、Kamei Katsuhiko、Kobayashi Masao、Okada Satoshi
  • Journal Title: Journal of Clinical Immunology Volume: - Issue: 5 Pages: 975-986
  • DOI: 10.1007/s10875-021-00988-7
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Mammalian VPS45 orchestrates trafficking through the endosomal system (2021)
  • Author(s): Frey Laura、Zietara Natalia、Lyszkiewicz Marcin、Marquardt Benjamin、Mizoguchi Yoko、Linder Monika I.、Liu Yanshan、Giesert Florian、Wurst Wolfgang、Dahlhoff Maik、Schneider Marlon R.、Wolf Eckhard、Somech Raz、Klein Christoph
  • Journal Title: Blood Volume: 137 Issue: 14 Pages: 1932-1944
  • DOI: 10.1182/blood.2020006871
• [Journal Article] Inborn errors of STAT1 immunity (2021)
  • Author(s): Mizoguchi Yoko、Okada Satoshi
  • Journal Title: Current Opinion in Immunology Volume: 72 Pages: 59-64
  • DOI: 10.1016/j.coi.2021.02.009
  • Peer Reviewed
• [Presentation] Successful donor marrow leukocyte infusion for cases with mixed chimerism following RIST from MRDs (2024)
  • Author(s): Yoko Mizoguchi
  • Organizer: 第46回日本造血・免疫細胞療法学会
• [Presentation] Sever congenital neutropenia (2023)
  • Author(s): Yoko Mizoguchi
  • Organizer: Klaus Betke Symposium 2023
• [Presentation] 小児血友病患者における関節症予防のために定期的な関節超音波評価の重要性 (2022)
  • Author(s): 溝口洋子
  • Organizer: 日本小児血液・がん学会
• [Presentation] 血友病個別化治療の実践に向けた当科の取り組み (2021)
  • Author(s): 溝口洋子、小林正夫
  • Organizer: 第43回日本血栓止血学会学術集会
  • Invited
• [Presentation] The efficacy of continuous US evaluation for joint health in pediatric patients with hemophilia. (2021)
  • Author(s): Yoko Mizoguchi, Chihiro Tani, Shiho Nishimura, Keiko Matsubara, Keita Tomioka, Maiko Shimomura, Yuko Nakashima, Hiroshi Kawaguchi, Satoshi Okada and Masao Kobayashi.
  • Organizer: 第83回日本血液学会学術集会
• [Presentation] 血友病診療の現在と未来 (2021)
  • Author(s): 溝口洋子
  • Organizer: 第73回 中国四国小児科学会
  • Invited
• [Book] 顆粒球減少症（好中球減少症）小児内科55巻増刊号 エキスパートが教える小児の 薬物治療 (2023)
  • Author(s): 溝口洋子
  • Total Pages: 4
  • Publisher: 東京医学社
• [Book] 重症先天性好中球減少症（SCN）. 原発性免疫不全症候群 診療の手引き 改訂第2版 自己免疫不全・自己炎症学会編集 (2023)
  • Author(s): 溝口洋子、岡田賢
  • Total Pages: 4
  • Publisher: 診断と治療社
• [Book] 好中球機能 小児臨床検査ガイド (2023)
  • Author(s): 溝口洋子、岡田賢
  • Total Pages: 4
  • Publisher: 文光堂