Elucidating the Function of Chemokine CCL20 in Inflammatory Bowel Disease using Genome-Edited Mouse Models

• Project/Area Number: 20K17025
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 53010:Gastroenterology-related
• Research Institution: Oita University
• Principal Investigator: Ariki Shimpei 大分大学, 医学部, 医員 (40849634)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2022: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: ケモカイン / 炎症性腸疾患 / ゲノム編集マウス / 疾患モデルマウス / CCL20 / CCR6 / CCR2

Outline of Research at the Start

炎症性腸疾患（IBD）は原因不明で再燃寛解を繰り返し慢性的に経過する難治性疾患である。近年Th17細胞、Treg細胞という新しいT細胞サブセットが見出され、IBDの病態形成との関係が指摘されている。Th17細胞、Treg細胞はケモカイン受容体CCR6を発現しており、細胞遊走因子であるケモカインCCL20に誘引されて組織へ集積してくるが、IBD病態形成に関するCCL20の生理学的役割は不明である。本研究では、当講座で作製したCCL20欠損マウスを用いて、定常状態におけるCCL20の腸管組織の恒常性に果たす生理学的役割を解明し、IBD病態モデルを作製および解析しCCL20の病態学的役割も解明する。

Outline of Final Research Achievements

To elucidate the physiological role of chemokines and chemokine receptors in inflammatory bowel disease, CCL20-, CCR2- and CCR6-deficient mice were generated by genome editing and analyzed for DSS-induced colitis; all mutant mice showed more severe colitis than wild-type mice. The frequencies of CD4+ T cells in the Peyer's patches were reduced in CCL20-deficient mice compared to those in wild-type mice after DSS treatment. In contrast, when CCR2/CCR6 double-deficient mice were generated, DSS-induced colitis was conversely milder than wild-type mice. We found that CCR2+CCR6+Th17 cells in the colonic lamina propria produced high levels of GM-CSF, suggesting that the migration of these inflammatory Th17 cells is important in the pathogenesis of colitis.

Academic Significance and Societal Importance of the Research Achievements

ダイナミックな免疫システムを理解するためには、白血球遊走を司るケモカインの役割を解明する必要がある。これまで、多発性硬化症や潰瘍性大腸炎の治療にα4インテグリン阻害薬が開発されており、今後もリンパ球のホーミングや遊走に関連した分子標的薬の開発が益々盛んになると考えられる。従って、ケモカインによる免疫制御機構の理解が自己免疫疾患の治療薬開発には欠かせないが、本研究により炎症性腸疾患におけるCCL20/CCR6およびCCR2の役割を分子生物学的手法を用いて個体レベルで明らかになったことは、学術的および社会的に意義が大きいと言える。

Research Products

• [Journal Article] GM-CSF-producing CCR2+ CCR6+ Th17 cells are pathogenic in dextran sodium sulfate-induced colitis model in mice (2023)
  • Author(s): Ariki Shimpei、Ozaka Sotaro、Sachi Nozomi、Chalalai Thanyakorn、Soga Yasuhiro、Fukuda Chiaki、Kagoshima Yomei、Ekronarongchai Supanuch、Mizukami Kazuhiro、Kamiyama Naganori、Murakami Kazunari、Kobayashi Takashi
  • Journal Title: Genes to Cells Volume: 28 Issue: 4 Pages: 267-276
  • DOI: 10.1111/gtc.13008
  • Peer Reviewed
• [Journal Article] CCL20/CCR6 chemokine signaling is not essential for pathogenesis in an experimental autoimmune encephalomyelitis mouse model of multiple sclerosis (2023)
  • Author(s): Sachi Nozomi、Kamiyama Naganori、Saechue Benjawan、Ozaka Sotaro、Dewayani Astri、Ariki Shimpei、Chalalai Thanyakorn、Soga Yasuhiro、Fukuda Chiaki、Kagoshima Yomei、Ekronarongchai Supanuch、Kobayashi Takashi
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 641 Pages: 123-131
  • DOI: 10.1016/j.bbrc.2022.11.088
  • Peer Reviewed
• [Journal Article] TRAF6 signaling pathway in T cells regulates anti-tumor immunity through the activation of tumor specific Th9 cells and CTLs (2022)
  • Author(s): Dewayani Astri、Kamiyama Naganori、Sachi Nozomi、Ozaka Sotaro、Saechue Benjawan、Ariki Shimpei、Goto Mizuki、Chalalai Thanyakorn、Soga Yasuhiro、Fukuda Chiaki、Kagoshima Yomei、Maekawa Yoichi、Kobayashi Takashi
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 613 Pages: 26-33
  • DOI: 10.1016/j.bbrc.2022.04.125
  • Peer Reviewed
• [Journal Article] Saireito, a Japanese herbal medicine, alleviates leaky gut associated with antibiotic-induced dysbiosis in mice (2022)
  • Author(s): Ozaka S、Sonoda A、Ariki S、Minata M、Kamiyama N、Hidano S、Sachi N、Ito K、Kudo Y、Dewayani A、Chalalai T、Ozaki T、Soga Y、Fukuda C、Mizukami K、Ishizawa S、Nishiyama M、Fujitsuka N、Mogami S、Kubota K、Murakami K、Kobayashi T
  • Journal Title: PLOS ONE Volume: 17 Issue: 6 Pages: e0269698-e0269698
  • DOI: 10.1371/journal.pone.0269698
  • Peer Reviewed / Open Access
• [Journal Article] Comprehensive lipidomics of lupus-prone mice using LC-MS/MS identifies the reduction of palmitoylethanolamide that suppresses TLR9-mediated inflammation. (2022)
  • Author(s): Ozaki T, Kamiyama N, Saechue B, Soga Y, Gotoh R, Nakayama T, Fukuda C, Dewayani A, Chalalai T, Ariki S, Ozaka S, Sonoda A, Hirose H, Gendo Y, Noguchi K, Sachi N, Hidano S, Maeshima K, Gotoh K, Masaki T, Ishii K, Osada Y, Shibata H, Kobayashi T
  • Journal Title: Genes Cells. Volume: 27 Issue: 7 Pages: 493-504
  • DOI: 10.1111/gtc.12944
  • Peer Reviewed / Open Access
• [Journal Article] Acute pancreatitis soon after COVID-19 vaccination (2022)
  • Author(s): Ozaka Sotaro、Kodera Takamoto、Ariki Shimpei、Kobayashi Takashi、Murakami Kazunari
  • Journal Title: Medicine Volume: 101 Issue: 2 Pages: e28471-e28471
  • DOI: 10.1097/md.0000000000028471
  • Peer Reviewed / Open Access
• [Journal Article] Analysis of the Prevalence and Species of <i>Anisakis</i> nematode in Sekisaba, <i>Scomber japonicus</i> Caught in Coastal Waters off Saganoseki, Oita in Japan (2021)
  • Author(s): Hidano Shinya、Mizukami Kazuhiro、Yahiro Takaaki、Shirakami Kohei、Ito Hideyuki、Ozaka Sotaro、Ariki Shimpei、Saechue Benjawan、Dewayani Astri、Chalalai Thanyakorn、Soga Yasuhiro、Goto Mizuki、Sonoda Akira、Ozaki Takashi、Sachi Nozomi、Kamiyama Naganori、Nishizono Akira、Murakami Kazunari、Kobayashi Takashi
  • Journal Title: Japanese Journal of Infectious Diseases Volume: 74 Issue: 5 Pages: 387-391
  • DOI: 10.7883/yoken.JJID.2020.859
  • NAID: 130008090913
  • ISSN: 1344-6304, 1884-2836
  • Year and Date: 2021-09-30
  • Peer Reviewed / Open Access
• [Journal Article] Protease inhibitory activity of secretory leukocyte protease inhibitor ameliorates murine experimental colitis by protecting the intestinal epithelial barrier (2021)
  • Author(s): Ozaka S, Sonoda A, Ariki S, Kamiyama N, Hidano S, Sachi N, Ito K, Kudo Y, Minata M, Saechue B, Dewayani A, Chalalai T, Soga Y, Takahashi Y, Fukuda C, Mizukami K, Okumura R, Kayama H, Murakami K, Takeda K, Kobayashi T.
  • Journal Title: Genes to Cells Volume: 26 Issue: 10 Pages: 807-822
  • DOI: 10.1111/gtc.12888
  • Peer Reviewed / Open Access
• [Journal Article] Monomorphic epitheliotropic intestinal T-cell lymphoma presenting as melena with long-term survival: A case report and review of literature (2021)
  • Author(s): Ozaka Sotaro、Inoue Kunimitsu、Okajima Tomoya、Tasaki Takako、Ariki Shimpei、Ono Hideki、Ando Takeaki、Daa Tsutomu、Murakami Kazunari
  • Journal Title: World Journal of Gastroenterology Volume: 27 Issue: 38 Pages: 6501-6510
  • DOI: 10.3748/wjg.v27.i38.6501
  • Peer Reviewed / Open Access
• [Journal Article] Development of a portable reverse transcription loop‐mediated isothermal amplification system to detect the E1 region of Chikungunya virus in a cost‐effective manner (2020)
  • Author(s): Saechue Benjawan、Kamiyama Naganori、Fukuda Chiaki、Watanabe Kei、Soga Yasuhiro、Goto Mizuki、Dewayani Astri、Ariki Shimpei、Hirose Haruna、Ozaka Sotaro、Sachi Nozomi、Hidano Shinya、Shirin Tahmina、Mondal Dinesh、Murakami Kazunari、Kobayashi Takashi
  • Journal Title: Genes to Cells Volume: 9 Issue: 9 Pages: 615-625
  • DOI: 10.1111/gtc.12797
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Identification of pathogenic Th17 cells in gastrointestinal tract using chemokine receptor deficient mice (2022)
  • Author(s): Shimpei Ariki, Sotaro Ozaka, Nozomi Sachi, Yomei Kagoshima, Thanyakorn Chalalai, Naganori Kamiyama, Takashi Kobayashi
  • Organizer: 第51回日本免疫学会学術集会
• [Presentation] Physiological function of chemokine CCL20 and its receptor CCR6 in multiple sclerosis in mice (2022)
  • Author(s): Nozomi Sachi, Naganori Kamiyama, Sotaro Ozaka, Shimpei Ariki, Thanyakorn Chalalai, Yomei Kagoshima, Takashi Kobayashi
  • Organizer: 第51回日本免疫学会学術集会
• [Presentation] Zika virus induces pathogenic T cell-attracting chemokines in the central nervous system and exacerbates experimental autoimmune encephalomyelitis (2022)
  • Author(s): Naganori Kamiyama, Thanyakorn Chalalai, Sotaro Ozaka, Shimpei Ariki, Yomei Kagoshima, Nozomi Sachi, Takashi Kobayashi
  • Organizer: 第51回日本免疫学会学術集会
• [Presentation] Secretory leukocyte protease inhibitor ameliorates murine experimental colitis by protecting the intestinal epithelial barrier (2022)
  • Author(s): Sotaro Ozaka, Shimpei Ariki, Yomei Kagoshima, Naganori Kamiyama, Nozomi Sachi, Thanyakorn Chalalai, Astri Dewayani, Takashi Kobayashi
  • Organizer: 第51回日本免疫学会学術集会
• [Presentation] Physiological relevance of TRAF6 signaling in dendritic cells in controlling C. rodentium infection (2022)
  • Author(s): Thanyakorn Chalalai, Naganori Kamiyama, Shimpei Ariki, Sotaro Ozaka, Nozomi Sachi, Yomei Kagoshima, Ryu Okumura, Kiyoshi Takeda, Takashi Kobayashi
  • Organizer: 第51回日本免疫学会学術集会
• [Presentation] Functional analysis of chemokine receptor CCR2 in a murine congenital toxoplasmosis model (2022)
  • Author(s): Noganori Kamiyama, Nozomi Sachi, Sotaro Ozaka, Shimpei Ariki, Thanyakorn Chalalai, Yasuhiro Soga, Chiaki Fukuda, Yomei Kagoshima, Supanuch Ekronarongchai, Masahiro Yamamoto, Takashi Kobayashi
  • Organizer: 第15回寄生虫感染免疫研究会
• [Presentation] Secretory leukocyte protease inhibitor ameliorates experimental colitis by protecting the intestinal epithelial barrier (2022)
  • Author(s): Sotaro Ozaka, Shimpei Ariki, Naganori Kamiyama, Yomei Kagoshima, Nozomi Sachi, Thanyakorn Chalalai, Yasuhiro Soga, Kazuhiro Mizukami, Kazunari Murakami, Takashi Kobayashi
  • Organizer: 第12回 癌・炎症と抗酸化研究会
• [Presentation] T細胞特異的TRAF6欠損マウスを用いた腸管におけるnon-pathogenic Th17細胞に関する研究 (2021)
  • Author(s): 有木晋平, 小坂聡太郎, 神山長慶, Benjawan Saechue, Astri Dewayani, Thanyakorn Chalalai, 佐知望美, 曽我泰裕, 福田千瑛, 水上一弘, 村上和成, 小林隆志
  • Organizer: 第44回日本分子生物学会年会
• [Presentation] CCL20欠損マウスを用いた多発性硬化症の発症におけるCCL20の機能解析 (2021)
  • Author(s): Nozomi Sachi, Naganori Kamiyama, Benjawan Saechue, Sotaro Ozaka, Astri Dewayani, Shimpei Ariki, Thanyakorn Chalalai, Yasuhiro Soga, Chiaki Fukuda, Takashi Kobayashi
  • Organizer: 第44回日本分子生物学会年会
• [Presentation] 大建中湯のSLPI発現増強を介した腸管保護作用の解析 (2021)
  • Author(s): 皆田美月, 小坂聡太郎, 有木晋平, 神山長慶, 佐知望美, Benjawan Saechue, Astri Dewayani, Thanyakorn Chalalai, 曽我泰裕, 福田千瑛, 水上一弘, 村上和成, 小林隆志
  • Organizer: 第44回日本分子生物学会年会
• [Presentation] 柴苓湯はマウスにおいて抗菌薬によるdysbiosisに起因するLeaky gutを軽減する (2021)
  • Author(s): 小坂聡太郎, 有木晋平, 皆田美月, 神山長慶，佐知望美，Benjawan Saechue，Astri Dewayani, Thanyakorn Chalalai, 曽我泰裕, 福田千瑛, 水上一弘, 村上和成, 小林隆志
  • Organizer: 第44回日本分子生物学会年会
• [Presentation] TRAF6 regulates Th9 cells and cytotoxic T lymphocytes in tumor immunity (2021)
  • Author(s): Astri Dewayani, Naganori Kamiyama, Shinya Hidano, Nozomi Sachi, Sotaro Ozaka, Shimpei Ariki, Benjawan Saechue, Yasuhiro Soga, Thanyakorn Chalalai, Chiaki Fukuda, Takashi Kobayashi
  • Organizer: 第44回日本分子生物学会年会
• [Presentation] Zika virus induces Th17 cell-attracting chemokines in the central nervous system and exacerbates neurological disorders. (2021)
  • Author(s): Naganori Kamiyama, Benjawan Saechue, Astri Dewayani, Chalalai Thanyakorn, Sotaro Ozaka, Shimpei Ariki, Chiaki Fukuda, Yasuhiro Soga, Mizuki Minata, Nozomi Sachi, Shinya Hidano, Takashi Kobayashi.
  • Organizer: 第44回日本分子生物学会年会
• [Presentation] Verification of T cell inhibitory effect of lipid mediator Oleylethanolamide (2021)
  • Author(s): Yasuhiro Soga, Naganori Kamiyama, Takashi Ozaki, Benjawan Saechue, Astri Dewayani, Nozomi Sachi, Sotaro Ozaka, Shimpei Ariki, Thanyakorn Chalalai, Chiaki Fukuda, Takashi Kobayash
  • Organizer: 第44回日本分子生物学会年会
• [Presentation] Identification of a responsible amino acid in prME protein for cell entry of Zika virus using single-round infectious particles (SRIPs) (2021)
  • Author(s): Chiaki Fukuda, Naganori Kamiyama, Benjawan Saechue, Astri Dewayani, Chalalai Thanyakorn, Sotaro Ozaka, Shimpei Ariki, Yasuhiro Soga, Mizuki Minata, Nozomi Sachi, Shinya Hidano, Ryosuke Suzuki,Takashi Kobayashi
  • Organizer: 第44回日本分子生物学会年会
• [Presentation] 樹状細胞におけるTRAF6シグナルがCitrobacter rodentium感染性腸炎に与える役割. (2021)
  • Author(s): 望月晶子, Thanyakorn chalalai, 伊藤加奈子, 有木 晋平, 小坂聡太郎, 佐知望美, 神山長慶, 小林隆志.
  • Organizer: 第11回 癌・炎症と抗酸化研究会(CIA研究会).
• [Presentation] マウス腸炎モデルを用いた大建中湯のSLPI発現を介した腸管保護作用の解析 (2021)
  • Author(s): 小坂聡太郎, 皆田美月, 有木晋平, 水上一弘, 神山長慶, 小林隆志, 村上和成.
  • Organizer: 第29回日本消化器関連学会週間 (JDDW 2021)
• [Presentation] 上部消化管出血を契機に診断され長期生存している単形性上皮向性腸管T細胞リンパ腫の1例 (2021)
  • Author(s): 小坂聡太郎, 井上邦光, 岡嶋智也, 田崎貴子, 有木晋平, 小野英樹, 村上和成.
  • Organizer: 第111回日本消化器内視鏡学会九州支部例会
• [Presentation] TRAF6 promotes the migration of Th17 cells into the CNS by regulating CCR6 expression in experimental autoimmune encephalomyelitis mice (2020)
  • Author(s): Naganori Kamiyama, Benjawan Saechue, Astri Dewayani, Chalalai Thanyakorn, Sotaro Ozaka, Shimpei Ariki, Yasuhiro Soga, Mizuki Goto, Nozomi Sachi, Shinya Hidano, Takashi Kobayashi
  • Organizer: 第43回日本分子生物学会年会
• [Presentation] TRAF6 Signaling in Th9 cells Regulates Anti-Tumor Immune Response (2020)
  • Author(s): Astri Dewayani, Naganori Kamiyama, Shinya Hidano, Nozomi Sachi, Sotaro Ozaka, Shimpei Ariki, Benjawan Saechue, Mizuki Goto, Yasuhiro Soga, Thanyakorn Chalalai, Takashi Kobayashi
  • Organizer: 第43回日本分子生物学会年会
• [Presentation] 抗生物質起因性大腸炎マウスモデルに対する柴苓湯の有効性の解明 (2020)
  • Author(s): 小坂 聡太郎, 園田 光, 有木 晋平, 水上 一弘, 村上 和成
  • Organizer: 第106回日本消化器病学会総会
• [Presentation] DSS腸炎モデルを用いたSLPIの結腸における生理的機能の解析 (2020)
  • Author(s): 小坂 聡太郎, 有木 晋平, 園田 光, 神山 長慶, 飛彈野 真也, 水上 一弘, 小林 隆志, 村上 和成
  • Organizer: 第28日本消化器関連学会週間(JDDW) 第62回日本消化器病学会大会