Molecular physiology of tight junction Claudin-family based on structural analysis

• Project/Area Number: 16H05121
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General physiology
• Research Institution: Osaka University
• Principal Investigator: Tamura Atsushi 大阪大学, 生命機能研究科, 特任准教授 (00362525)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000); Fiscal Year 2018: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2017: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2016: ¥6,630,000 (Direct Cost: ¥5,100,000、Indirect Cost: ¥1,530,000)
• Keywords: クローディン / タイトジャンクション / 上皮細胞 / 細胞間接着装置 / 上皮細胞間バリア / 構造解析 / 炎症 / 生体機能 / 分子動力学 / 微小環境 / ノックアウトマウス / 細胞間接着 / 細胞間バリア / 遺伝子改変マウス / ストランド / ノックアウト / 分子生理学

Outline of Final Research Achievements

Epithelial cells are the largest number of cells in the body that cover the internal and external surfaces of multicellular organisms. They are responsible for the functions and structures of major organs such as the skin, liver, and kidneys. The TJ (Tight Junction), composed of the polymer of claudin (Cldn) family protein molecules, reside in between epithelial cells. The paracellular permeability is mainly determined to maintain the tissue microenvironment properly. Biological functions are carried out　within the exactly regulated tissue microenvironment. The failure of it is linked to various pathological conditions directly. In this application, we generate the Cldn null epithelial cells by using the CRISPR/CAS9 system to advance the functional analysis of Cldn-based TJ.

Academic Significance and Societal Importance of the Research Achievements

多細胞生物は、隣り合う上皮細胞同士が接着することで形成された上皮細胞シートにより、体内環境と体外環境が隔絶されることで、体内環境を維持する。このとき、適切に制御された上皮細胞シートバリアが必要である。上皮細胞シートバリアが重度に障害されると、脱水や生命に必要な酵素活性低下し生命も脅かされる。より軽度であっても、炎症や栄養分吸収障害などの機能障害を惹起する場合がある。申請者は、上皮細胞シートバリアの中でも、特に、上皮細胞間バリアに関わるタイトジャンクションおよびその構成タンパク質クローディンの構造と機能解析を進めている。胃酸と炎症など、今後の展開により疾患の治療と予防につながる成果が得られた。

Research Products

• [Int'l Joint Research] ハーバード大学/病理学教室(米国)
• [Journal Article] Reciprocal Association between the Apical Junctional Complex and AMPK: A Promising Therapeutic Target for Epithelial/Endothelial Barrier Function? (2019)
  • Author(s): Tsukita, K., Yano, T., Tamura, A., and Tsukita, S.
  • Journal Title: Int J Mol Sci. Volume: 20 Issue: 23 Pages: 6012-6012
  • DOI: 10.3390/ijms20236012
  • Peer Reviewed / Open Access
• [Journal Article] Vinculin is critical for the robustness of the epithelial cell sheet paracellular barrier for ions. (2019)
  • Author(s): Konishi S, Yano T, Tanaka H, Mizuno T, Kanoh H, Tsukita K, Namba T, Tamura A, Yonemura S, Gotoh S, Matsumoto H, Hirai T, Tsukita S.
  • Journal Title: Life Sci Alliance Volume: 2 Issue: 4 Pages: e201900414-e201900414
  • DOI: 10.26508/lsa.201900414
  • NAID: 120007144472
  • Peer Reviewed / Open Access
• [Journal Article] Deficiency of Stomach-Type Claudin-18 in Mice Induces Gastric Tumor Formation Independent of H. Pylori Infection. (2019)
  • Author(s): Suzuki K, Sentani K, Tanaka H, Yano T, Suzuki K, Oshima M, Yasui W, Tamura A, Tsukita S.
  • Journal Title: Cell Mol Gastroenterol Hepatol. Volume: 印刷中 Issue: 1 Pages: 119-142
  • DOI: 10.1016/j.jcmgh.2019.03.003
  • Peer Reviewed / Open Access
• [Journal Article] Morphologic determinant of tight junctions revealed by claudin-3 structures (2019)
  • Author(s): Nakamura Shun、Irie Katsumasa、Tanaka Hiroo、Nishikawa Kouki、Suzuki Hiroshi、Saitoh Yasunori、Tamura Atsushi、Tsukita Sachiko、Fujiyoshi Yoshinori
  • Journal Title: Nature Communications Volume: 10 Issue: 1 Pages: 816-816
  • DOI: 10.1038/s41467-019-08760-7
  • Peer Reviewed / Open Access
• [Journal Article] Interleukin 22 Expands Transit-Amplifying Cells While Depleting Lgr5(+) Stem Cells via Inhibition of Wnt and Notch Signaling. (2019)
  • Author(s): Zha JM, Li HS, Lin Q, Kuo WT, Jiang ZH, Tsai PY, Ding N, Wu J, Xu SF, Wang YT, Pan J, Zhou XM, Chen K, Tao M, Odenwald MA, Tamura A, Tsukita S, Turner JR, He WQ. Interleukin 22 Expands Transit-Amplifying Cells While Depleting Lgr5(+) Stem Cells via Inhibition of Wnt and Notch Signaling.
  • Journal Title: Cell Mol Gastroenterol Hepatol. Volume: 7 Issue: 2 Pages: 255-274
  • DOI: 10.1016/j.jcmgh.2018.09.006
  • Peer Reviewed / Open Access
• [Journal Article] The Claudins: From Tight Junctions to Biological Systems (2019)
  • Author(s): Tsukita Sachiko、Tanaka Hiroo、Tamura Atsushi
  • Journal Title: Trends in Biochemical Sciences Volume: 44 Issue: 2 Pages: 141-152
  • DOI: 10.1016/j.tibs.2018.09.008
  • Peer Reviewed
• [Journal Article] Claudin-3 regulates bile canalicular paracellular barrier and cholesterol gallstone core formation in mice. (2019)
  • Author(s): Tanaka H, Imasato M, Yamazaki Y, Matsumoto K, Kunimoto K, Delpierre J, Meyer K, Zerial M, Kitamura N, Watanabe M, Tamura A, Tsukita S.
  • Journal Title: J Hepatol. Volume: 69 Issue: 6 Pages: 1308-1316
  • DOI: 10.1016/j.jhep.2018.08.025
  • Peer Reviewed
• [Journal Article] Claudin-3 Loss Causes Leakage of Sweat from the Sweat Gland to Contribute to the Pathogenesis of Atopic Dermatitis (2017)
  • Author(s): Yamaga Kosuke、Murota Hiroyuki、Tamura Atsushi、Miyata Hirofumi、Ohmi Masato、Kikuta Junichi、Ishii Masaru、Tsukita Sachiko、Katayama Ichiro
  • Journal Title: Journal of Investigative Dermatology Volume: - Issue: 6 Pages: 1279-1287
  • DOI: 10.1016/j.jid.2017.11.040
  • Peer Reviewed
• [Journal Article] Site-specific distribution of claudin-based paracellular channels with roles in biological fluid flow and metabolism (2017)
  • Author(s): Tanaka Hiroo、Tamura Atsushi、Suzuki Koya、Tsukita Sachiko
  • Journal Title: Annals of the New York Academy of Sciences Volume: 1405 Issue: 1 Pages: 44-52
  • DOI: 10.1111/nyas.13438
  • Peer Reviewed
• [Journal Article] Apical cytoskeletons and junctional complexes as a combined system in epithelial cell sheets (2017)
  • Author(s): Yano Tomoki、Kanoh Hatsuho、Tamura Atsushi、Tsukita Sachiko
  • Journal Title: Annals of the New York Academy of Sciences Volume: 1405 Issue: 1 Pages: 32-43
  • DOI: 10.1111/nyas.13432
  • Peer Reviewed
• [Journal Article] Time- and dose-dependent claudin contribution to biological functions: Lessons from claudin-1 in skin (2017)
  • Author(s): Tokumasu Reitaro、Tamura Atsushi、Tsukita Sachiko
  • Journal Title: Tissue Barriers Volume: 5 Issue: 3 Pages: e1336194-e1336194
  • DOI: 10.1080/21688370.2017.1336194
  • Peer Reviewed
• [Journal Article] IL-22?Upregulates Epithelial Claudin-2 to Drive Diarrhea and Enteric Pathogen Clearance (2017)
  • Author(s): Tsai Pei-Yun、Zhang Bingkun、He Wei-Qi、Zha Juan-Min、Odenwald Matthew A.、Singh Gurminder、Tamura Atsushi、Shen Le、Sailer Anne、Yeruva Sunil、Kuo Wei-Ting、Fu Yang-Xin、Tsukita Sachiko、Turner Jerrold R.
  • Journal Title: Cell Host & Microbe Volume: 21 Issue: 6 Pages: 671-681.e4
  • DOI: 10.1016/j.chom.2017.05.009
  • Peer Reviewed
• [Journal Article] Dose-dependent role of claudin-1 in vivo in orchestrating features of atopic dermatitis. (2016)
  • Author(s): Tokumasu R, Yamaga K, Yamazaki Y, Murota H, Suzuki K, Tamura A, Bando K, Furuta Y, Katayama I, Tsukita S.
  • Journal Title: Proc Natl Acad Sci U S A. Volume: 113 Issue: 28
  • DOI: 10.1073/pnas.1525474113
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] タイトジャンクション - クローディンによる細胞間バリア制御と、その異 常と病態ークローディン欠損マウス解析の視点からー (2019)
  • Author(s): 田村 淳、田中 啓雄、 矢野 智樹、鈴木 浩也、 月田 早智子
  • Organizer: 第71回日本細胞生物学会大会/ 第19回日本蛋白質科学会年会 合同大会
• [Presentation] Role of planar cell polarity-dependent distribution of apical microtubules in the coordinated ciliary beating of tracheal multiciliated cells. (2018)
  • Author(s): Shogo Nakayama, Elisa Herawati, Maki Takagishi, Takayuki Torisawa, Toshinori Namba, Tomoki Yano, Atsushi Tamura, Shuji Ishihara, Kazuhiro Oiwa, Masahide Takahashi, Sachiko Tsukita
  • Organizer: ASCB 2018 Annual Meeting
• [Presentation] TJ-MAP4 up-regulates the level of ppMLC at the circumferential ring for epithelial apical constriction (2018)
  • Author(s): Hatsuho Kanoh, Tomoki Yano, Atsushi Tamura, Yuhei Goto, Shogo Nakayama, Akira Komatsubara, Kazuhiro Aoki and Sachiko Tsukita
  • Organizer: 第91回日本生化学会大会
• [Presentation] Continuous Gastric Acid Leakage due to Dysfunctional Paracellular-Barrier Function Causes Gastric Tumor via CXCL5 and Wnt Signaling with Aging (2018)
  • Author(s): Koya Suzuki, Kazuhiro Sentani, Hiroo Tanaka, Tomoki Yano, Kazuo Suzuki, Atsushi Tamura, Sachiko Tsukita,
  • Organizer: 第91回日本生化学会大会
• [Presentation] Establishment of a PCP-dependent apical microtubule network in tracheal MCCs (2018)
  • Author(s): Shogo Nakayama, Elisa Herawati, Maki Takagishi, Tomoki Nishida, Kanako Inoue, Takayuki Torisawa, Toshinori Namba, Shuji Ishihara, Hiroo Tanaka, Tomoki Yano, Atsushi Tamura, Kazuhiro Oiwa, Masahide Takahashi, SachikoTsukita
  • Organizer: Joint Annual Meeting Of JSDB 51st and JSCB 70th(第70回日本細胞生物学会・第53回日本発生生物学会合同大会)
• [Presentation] Tight junctional cingulin organizes the apical intermediate filaments (2018)
  • Author(s): Yuki Nakao, Hiroo Tanaka, Tomohiro Tamura, Shogo Nakayama, Akira Yamamoto, Hatsuho Kanoh, Atsushi Tamura, Tomoki Yano, Sachiko Tsukita
  • Organizer: Joint Annual Meeting Of JSDB 51st and JSCB 70th(第70回日本細胞生物学会・第53回日本発生生物学会合同大会)
• [Presentation] 上皮細胞シート形態形成におけるタイトジャンクション微小管結合タンパク質の解析 (2017)
  • Author(s): 加納初穂、矢野智樹、氏昌未、田村朋宏、田村淳、月田早智子
  • Organizer: 第69回 日本細胞生物学会大会
• [Presentation] クローディン15における細胞間チャネル機能のイオン選択性とサイズ選択性制御機構の検討 (2017)
  • Author(s): 鈴木浩也、武知真希、谷一寿、田村淳、藤吉好則、月田早智子
  • Organizer: 第69回 日本細胞生物学会大会
• [Presentation] 気管多繊毛上皮細胞における繊毛配列動態 (2017)
  • Author(s): 月田早智子、加納初穂、Elisa Herawati、小西聡史、立石和博、 矢野智樹、田村淳
  • Organizer: 第69回 日本細胞生物学会大会
• [Presentation] タイトジャンクション構成タンパク質クローディンが規定する生体微小環境恒常性と消化管機能 (2017)
  • Author(s): 田村淳、田中啓雄、高橋信之、月田早智子
  • Organizer: 日本農芸化学会大会
  • Place of Presentation: 京都女子大
  • Invited
• [Presentation] Bile flow and gallstone diseases induced by the deficiency of claudin family proteins (2016)
  • Author(s): Atsushi Tamura
  • Organizer: International conference Tight junction and their proteins
  • Place of Presentation: Germany
  • Int'l Joint Research / Invited
• [Remarks] 大阪大学大学院生命機能研究科
  • URL: http://www.fbs.osaka-u.ac.jp/
• [Remarks] 大阪大学大学院生命機能研究科 月田研究室
  • URL: http://www.fbs.osaka-u.ac.jp/labs/tsukita/
• [Remarks] 大阪大学月田研究室
  • URL: http://www.fbs.osaka-u.ac.jp/labs/tsukita/
• [Remarks] 大阪大学生命機能研究科
  • URL: http://www.fbs.osaka-u.ac.jp/
• [Remarks] 大阪大学医学系研究科
  • URL: http://www.med.osaka-u.ac.jp/
• [Remarks] 大阪大学大学院生命機能研究科/医学系研究科 月田研究室
  • URL: http://www.fbs.osaka-u.ac.jp/labs/tsukita/