Physiological roles of Phosphoinositide-specific phosphatase in lung development and maturation through regulating membrane trafficking.

• Project/Area Number: 17K08532
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General physiology
• Research Institution: Kanazawa University
• Principal Investigator: Yoshioka Kazuaki 金沢大学, 医学系, 講師 (80333368)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2019: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2017: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: ホスホイノシタイド / 脂質ホスファターゼ / ミオチュブラリン関連タンパク質 / PI3キナーゼ / エンドサイトーシス / オートファジー / リソソーム / II型肺胞上皮細胞 / エンドソーム / 細胞内クリアランス / ホスファチジルイノシトール / ミオチュブラリン / 肺胞 / 肺線維化 / ホスホイノシチド / 細胞内小胞輸送 / MTMR4 / イノシトールリン脂質 / クラスII型PI3キナーゼ / 肺上皮細胞 / 細胞内膜輸送 / ノックアウトマウス

Outline of Final Research Achievements

Phosphatidylinositol 3-phosphate (PI(3)P) is the predominant phosphoinositide species in early endosomes and autophagosomes, in which PI(3)P dictates traffic of these organelles. Phosphoinositide levels are tightly regulated by lipid-kinases and-phosphatases; however, a phosphatase that converts PI(3)P back to phosphatidylinositol in the endosomal and autophagosomal compartments is not fully understood. We investigated the subcellular distribution and functions of myotubularin-related protein-4 (MTMR4), which is distinct among other MTMRs in that it possesses a PI(3)P-binding FYVE domain, in lung alveolar epithelial cells. MTMR4 knockdown markedly suppressed the motility, fusion, and fission of PI(3)P-enriched structures, resulting in decreases in late endosomes, autophagosomes, and lysosomes. This project has unveiled that MTMR4 is essential for the integrity of endocytic and autophagic pathways.

Academic Significance and Societal Importance of the Research Achievements

サーファクタントを分泌するII型肺胞上皮（AECII）細胞は正常肺発生において肺胞を形成する組織幹細胞であり、また外部環境由来の病原体アラームシグナルをいち早く感知し、これらによる肺組織の障害に対する防御・修復応答に中心的な役割をはたすことが明らかになってきた。本研究の成果は、MTMR4がAECII細胞の細胞内クリアランス制御を介して細胞分化・増殖を調整して肺組織の構築および恒常性を維持する機能を有することを示した。この研究成果は肺形成・発達メカニズムの解明のみならず、肺線維症等の炎症性肺疾患（全世界の死亡原因第３）の病態解明と新しい治療法開発戦略において有用な情報を与える。

Research Products

• [Journal Article] TGFβ receptor endocytosis and Smad signaling require synaptojanin1?, PI3K-C2α?, and INPP4B?mediated phosphoinositide conversions (2020)
  • Author(s): Aki Sho、Yoshioka Kazuaki、Takuwa Noriko、Takuwa Yoh
  • Journal Title: Molecular Biology of the Cell Volume: 31(5) Issue: 5 Pages: 360-372
  • DOI: 10.1091/mbc.e19-11-0662
  • Peer Reviewed / Open Access
• [Journal Article] Class II phosphatidylinositol 3-kinase α and β isoforms are required for vascular smooth muscle Rho activation, contraction and blood pressure regulation in mice (2020)
  • Author(s): Islam Shahidul、Yoshioka Kazuaki、Aki Sho、Ishimaru Kazuhiro、Yamada Hiroki、Takuwa Noriko、Takuwa Yoh
  • Journal Title: The Journal of Physiological Sciences Volume: 70 Issue: 1 Pages: 18-18
  • DOI: 10.1186/s12576-020-00745-2
  • Peer Reviewed / Open Access
• [Journal Article] The class II phosphoinositide 3-kinases PI3K-C2α and PI3K-C2β differentially regulate clathrin-dependent pinocytosis in human vascular endothelial cells. (2019)
  • Author(s): Aung KT, Yoshioka K, Ishimaru K, Aki S, Takuwa N, and Takuwa Y.
  • Journal Title: Journal of Physiological Sciences Volume: 69 Issue: 2 Pages: 263-280
  • DOI: 10.1007/s12576-018-0644-2
  • Peer Reviewed / Open Access
• [Journal Article] Sphigosine-1-phosphate receptor 1 promotes neointimal hyperplasia in a mouse model of carotid artery injury. (2019)
  • Author(s): Kitano T, Takashima S, Inoue O, Goten C, Nomura A, Yoshioka K, Okajima M, Kaneko S, Takuwa Y, Takamura M.
  • Journal Title: Biochem. Biphys. Res. Communi. Volume: 511 Issue: 1 Pages: 179-184
  • DOI: 10.1016/j.bbrc.2019.02.047
  • Peer Reviewed / Open Access
• [Journal Article] Lysophosphatidic acid?induced YAP/TAZ activation promotes developmental angiogenesis by repressing Notch ligand Dll4 (2019)
  • Author(s): Yasuda Daisuke、Kobayashi Daiki、Akahoshi Noriyuki、Ohto-Nakanishi Takayo、Yoshioka Kazuaki、Takuwa Yoh、Mizuno Seiya、Takahashi Satoru、Ishii Satoshi
  • Journal Title: Journal of Clinical Investigation Volume: 129 Issue: 10 Pages: 4332-4349
  • DOI: 10.1172/jci121955
  • Peer Reviewed / Open Access
• [Journal Article] Sphingosine kinase-2 prevents macrophage cholesterol accumulation and atherosclerosis by stimulating autophagic lipid degradation (2019)
  • Author(s): Ishimaru Kazuhiro、Yoshioka Kazuaki、Kano Kuniyuki、Kurano Makoto、Saigusa Daisuke、Aoki Junken、Yatomi Yutaka、Takuwa Noriko、Okamoto Yasuo、Proia Richard L.、Takuwa Yoh
  • Journal Title: Scientific Reports Volume: 9 Issue: 1 Pages: 18329-18329
  • DOI: 10.1038/s41598-019-54877-6
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Class II PI3Ks α nad β are required for Rho-dependent uterine smooth muscle contraction and parturition in mice. (2019)
  • Author(s): Sarker AK, Aki S, Yoshioka K, Kuno K, Okamoto Y, Ishimaru K, Takuwa N, and Takuwa Y.
  • Journal Title: Endocrinology Volume: 160 Issue: 1 Pages: 235-248
  • DOI: 10.1210/en.2018-00756
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] MTMR4, a phosphoinositide specific 3’-phosphatase, regulates TFEB activity and the endocytic and autophagic pathways. (2018)
  • Author(s): Pham HQ, Yoshioka K, Mohri H, Nakata H, Aki S, Ishimaru K, Takuwa N, and Takuwa Y.
  • Journal Title: Genes to Cells Volume: 23 Issue: 8 Pages: 670-687
  • DOI: 10.1111/gtc.12609
  • NAID: 120006621947
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Photoacoustic imaging of tumor vascular permeability with indocyanine green in a mouse model. (2018)
  • Author(s): Okumura K, Yoshida K, Yoshioka K, Aki S, Yoneda N, Inoue D, Kitao A, Ogi T, Kozaka K, Minami T, Koda W, Kobayashi S, Takuwa Y and Gabata T
  • Journal Title: European Radiology Experimental Volume: 2: 5 Issue: 1 Pages: 1-9
  • DOI: 10.1186/s41747-018-0036-7
  • Peer Reviewed / Open Access
• [Journal Article] Augmented sphingosine 1 phosphate receptor-1 signaling in cardiac fibroblasts induces cardiac hypertrophy and fibrosis through angiotensin II and interleukin-6. (2017)
  • Author(s): Ohkura SI, Usui S, Takashima SI, Takuwa N, Yoshioka K, Okamoto Y, Inagaki Y, Sugimoto N, Kitano T, Takamura M, Wada T, Kaneko S, Takuwa Y.
  • Journal Title: PLoS One. Volume: 12 Issue: 8 Pages: e0182329-e0182329
  • DOI: 10.1371/journal.pone.0182329
  • Peer Reviewed / Open Access
• [Journal Article] Early endothelialization associated with a biolimus A9 bioresorbable polymer stent in a porcine coronary model. (2017)
  • Author(s): Mori M, Sakata K, Nakanishi C, Nakahashi T, Kawashiri MA, Yoshioka K, Takuwa Y, Okada H, Yokawa JI, Shimojima M, Yoshimuta T, Yoshida S, Yamagishi M, Hayashi K.
  • Journal Title: Heart Vessels. Volume: 32 Issue: 10 Pages: 1244-1252
  • DOI: 10.1007/s00380-017-0992-5
  • Peer Reviewed
• [Journal Article] Pancreas lineage allocation and specification are regulated by sphingosine-1-phosphate signalling. (2017)
  • Author(s): Serafimidis I, Rodriguez-Aznar E, Lesche M, Yoshioka K, Takuwa Y, Dahl A, Pan D and Gavalas A.
  • Journal Title: PLOS Biology Volume: 15 (3) Issue: 3 Pages: e2000949-e2000949
  • DOI: 10.1371/journal.pbio.2000949
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Differential roles of class II PI3-kinase-C2α and -C2β in clathrinmediated fluid phase endocytosis in vascular endothelial cells (2020)
  • Author(s): Kazuaki Yoshioka, Sho Aki, Noriko Takuwa, Yoh Takuwa
  • Organizer: 第９７回 日本生理学会大会（大分・別府）
• [Presentation] Sphingosine kinase-2 is required for autophagic lipid degradation in macrophage and inhibits atherosclerosis (2020)
  • Author(s): Yoh Takuwa, Kazuhiro Ishimaru, Kazuaki Yoshioka, Noriko Takuwa, Yasuo Okamoto
  • Organizer: 第９７回 日本生理学会大会（大分・別府）
• [Presentation] Sequential phosphoinositide conversion is required for TGFβ-induced receptor endocytosis and endosomal receptor signaling in endothelial cells (2020)
  • Author(s): Sho Aki, Kazuaki Yoshioka, Noriko Takuwa, Yoh Takuwa
  • Organizer: 第９７回 日本生理学会大会（大分・別府）
• [Presentation] 血管内皮細胞のDll4-Notch シグナルにおけるクラスII型PI3 キナーゼ-C2α の機能 (2019)
  • Author(s): 清水奨太、吉岡和晃、安藝 翔、多久和 陽
  • Organizer: 日本生化学会北陸支部第37回大会、ハルピン・ホール（福井）
• [Presentation] ノシトールリン脂質3’-ホスファターゼMTMR4によるエンド-リソソームおよびオートファジー経路の制御メカニズム (2019)
  • Author(s): 吉岡和晃、Quynh Hoa Pham、安藝 翔、多久和 陽
  • Organizer: 日本生化学会北陸支部第37回大会、ハルピン・ホール（福井）
• [Presentation] Sequential phosphoinositide conversion is required for transforming growth factor β-induced receptor endocytosis and Smad2/3 activation in endothelial cells (2019)
  • Author(s): Sho Aki, Kazuaki Yoshioka, Noriko Takuwa, Yoh Takuwa
  • Organizer: 60th International Conference on the Bioscience of Lipids, Hitotsubashi Hall (Tokyo)
  • Int'l Joint Research
• [Presentation] Differential roles of class II PI3K in endocytosis and endosomal signaling (2019)
  • Author(s): Kazuaki Yoshioka, Khin Thuzar Aung, Md Azadul Kabir Sarker, Sho Aki, Kuntal Biswas, Noriko Takuwa, Yoh Takuwa
  • Organizer: 60th International Conference on the Bioscience of Lipids, Hitotsubashi Hall (Tokyo)
  • Int'l Joint Research
• [Presentation] 連続するホスホイノシタイド代謝がTGFβ受容体エンドサイトーシス及びSmad2/3 活性化に必須である (2019)
  • Author(s): 安藝 翔、吉岡和晃、多久和典子、多久和 陽
  • Organizer: 第61回 日本脂質生化学会、北海道大学（札幌）
• [Presentation] イノシトールリン脂質3’脱リン酸化酵素MTMR4 による細胞内クリアランス制御メカニズム (2019)
  • Author(s): 吉岡和晃、Quynh Hoa Pham、安藝 翔、多久和典子、多久和 陽
  • Organizer: 第61回 日本脂質生化学会、北海道大学（札幌）
• [Presentation] 連続するホスホイノシタイド代謝がTGFβ受容体エンドサイトーシス及びSmad2/3活性化に必須である (2019)
  • Author(s): 安藝 翔、吉岡和晃、Islam Shahidul、多久和典子、多久和 陽
  • Organizer: 第92回日本生化学会大会、横浜パシフィコ（横浜）
• [Presentation] 細胞膜イノシトールリン脂質脱リン酸化酵素・MTMR4による細胞内クリアランスの制御 (2019)
  • Author(s): 吉岡和晃、Quynh Hoa Pham、安藝 翔、多久和典子、多久和 陽
  • Organizer: 第92回日本生化学会大会、横浜パシフィコ（横浜）
• [Presentation] Essential role of class II PI3K in endocytosis and endosomal signaling. (2019)
  • Author(s): Yoshioka, K., Aung, KT., Sarker, Md.A.K., Aki, S., Biswas, K., Takuwa, N., Takuwa, Y.
  • Organizer: 9th Federation of the Asia and Oceanian Physiological Societies (FAOPS2019)
  • Int'l Joint Research / Invited
• [Presentation] Sequential phosphoinositide conversion is required for TGFβ-induced receptor endocytosis in ECs. (2019)
  • Author(s): Aki, S., Yoshioka, K., Takuwa, N., Takuwa, Y.
  • Organizer: 9th Federation of the Asia and Oceanian Physiological Societies (FAOPS2019)
  • Int'l Joint Research
• [Presentation] Myotubularin-related protein 4 (MTMR4), a phosphoinositide 3'-phosphatase, regulates endolysosome integrity and autophagy flux in human lung alveolar epithelial A549 cells (2018)
  • Author(s): Pham Quynh Hoa, Yoshioka Kazuaki, Azadul Sarker Kabir, Aung Thuzar Khin, Islam Shahidul, Aki Sho, Ishimaru Kazuhiro, Takuwa Noriko, Takuwa Yoh
  • Organizer: 第 95 回日本生理学会大会
• [Presentation] Indispensable role of α and β isoforms of class II phosphoinositide 3-kinases (PI3K) in the uterine smooth muscle contraction during labor (2018)
  • Author(s): Sarker Kabir Azadul, Sho Aki, Yoshioka Kazuaki, Kuno Koji, Okamoto Yasuo, Aung Thuzar Khin, Pham Quynh Hoa, Islam Shahidul, Takuwa Noriko, Takuwa Yoh
  • Organizer: 第 95 回日本生理学会大会
• [Presentation] Class II phosphoinositide 3-kinase isoforms PI3K-C2αand PI3K-C2β play essential roles in pinocytosis in vascular endothelial cells. (2018)
  • Author(s): Aung Thuzar Khin, Yoshioka Kazuaki, Aki Sho, Pham Quynh Hoa, Sarker Kabir Azadul, Islam Shahidul, Ishimaru Kazuhiro, Takuwa Noriko, Takuwa Yoh
  • Organizer: 第 95 回日本生理学会大会
• [Presentation] Myotubularin-related protein 4 (MTMR4), a phosphoinositide 3'-speciphosphatase, regulates endolysosome integrity and autophagy in A549 human lung carcinoma cells. (2017)
  • Author(s): Quynh Hoa Pham , Kazuaki Yoshioka , Azadul Kabir Sarker , Khin Thuzar Aung , Shahidul Islam , Sho Aki , Sato Nakamura , Noriko Takuwa, Yoh Takuwa
  • Organizer: 第40回日本分子生物学会年会