Metabolic features of hormone secreting cells in pancreatic islets and analyses of their dysfunction using pseudo-islets

• Project/Area Number: 20K08918
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 54040:Metabolism and endocrinology-related
• Research Institution: Nihon University
• Principal Investigator: ISHIHARA Hisamitsu 日本大学, 医学部, 教授 (60361086)
• Co-Investigator(Kenkyū-buntansha): 小須田 南 日本大学, 医学部, 助手 (40811609)
• 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: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2021: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: pancreatic beta-cells / pancreatic alpha-cells / glucose metabolism / metabolic reprogramming / インスリン分泌 / グルカゴン分泌 / 遺伝子発現修飾 / 偽膵島 / 2型糖尿病 / 膵ランゲルハンス島 / インスリン / グルカゴン

Outline of Research at the Start

糖尿病の約9割は、2型糖尿病と呼ばれる生活習慣の問題が大きく影響する病気です。その原因は、膵臓にある細胞の塊である膵ランゲルハンス島（別名膵島）が障害されることです。膵島には、血糖値を下げるインスリンを分泌する細胞と血糖値を上昇させるグルカゴンを分泌する細胞があります。本研究では、インスリン分泌細胞とグルカゴン分泌細胞に類似した2種類の培養細胞を用います。血糖値が高くなるとインスリンやグルカゴンの分泌が変化しますが、2型糖尿病では適切に分泌変化が起こらなくなっています。これらの細胞に遺伝子操作を施し、どこがおかしくなると糖尿病のようになるかを、突き止めることを目指してこの研究を行います。

Outline of Final Research Achievements

By analyzing effects of modulation of expression levels of several enzymes constituting the TCA cycle in insulin secreting MIN6 cells, we have suggested that glutamate is important for the metabolism-secretion coupling in insulin secretion evoked by glucose. We have also identified Sox11 as a transcription factor regulating general glucose metabolism in insulin secreting cells. Since Sox11 is known to be a factor important for the epithelial mesenchymal transition, Sox11 may be important for metabolic reprogramming in cell fate determination in insulin secreting cells.
In addition, we successfully generated pseudo-islets from insulin secreting MIN6 cells and glucagon secreting aTC1 cells. Our analyses suggested that the two cell types establish a specific configuration in the cell clusters, which may be important for their hormone secreting properties.

Academic Significance and Societal Importance of the Research Achievements

本研究の成果は、膵β細胞のインスリン分泌機構の解明を進めるもので、ひいてはインスリン分泌を増強させる薬剤の開発につながる。インスリン分泌を増強する場合には、β細胞の疲弊をもたらさないことが重要であるが、本研究ではインスリン分泌と細胞の運命の両者を制御する因子の候補が見出された。このような因子の存在は、β細胞の疲弊無しにインスリン分泌を増強する薬剤の開発の第一歩となるものである。このターゲットを修飾する食品成分の同定ができれば、2型糖尿病発症・進展予防の食事戦略の開発にもつながる。さらに、偽膵島を用いた解析は、複数の構成細胞からなる膵島のホルモン分泌メカニズムの解明に展開できると考えられる。

Research Products

• [Journal Article] A large-scale functional analysis of genes expressed differentially in insulin secreting MIN6 sublines with high versus mildly reduced glucose-responsiveness, (2023)
  • Author(s): Tanaka A, Kosuda M, Yamana M, Furukawa A, Nagasawa A, Fujishiro M, Kohno G, Ishihara H.
  • Journal Title: Scientific Reports Volume: 13 Issue: 1 Pages: 5654-5654
  • DOI: 10.1038/s41598-023-32589-2
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Identification of serum metabolome signatures associated with retinal and renal complications of type 2 diabetes. (2023)
  • Author(s): Tomofuji Y, Suzuki K, Kishikawa T, Shojima N, Hosoe J, Inagaki K, Matsubayashi S, Ishihara H, Watada H, Ishigaki Y; BioBank Japan Project; Inohara H, Murakami Y, Matsuda K, Okada Y, Yamauchi T, Kadowaki T
  • Journal Title: Communications Medicine Volume: 3 Issue: 1 Pages: 5-5
  • DOI: 10.1038/s43856-022-00231-3
  • Peer Reviewed / Open Access
• [Journal Article] Glucagon Response to Glucose Challenge in Patients with Idiopathic Postprandial Syndrome (2022)
  • Author(s): Kosuda M, Watanabe K, Koike M, Morikawa A, Saito H, Kohno G, Ishihara H.
  • Journal Title: Journal of Nippon Medical School Volume: 89 Issue: 1 Pages: 102-107
  • DOI: 10.1272/jnms.JNMS.2022_89-205
  • NAID: 130008087911
  • ISSN: 1345-4676, 1347-3409
  • Year and Date: 2022-02-25
  • Peer Reviewed / Open Access
• [Journal Article] Metabolism-secretion coupling in glucose-stimulated insulin secretion (2022)
  • Author(s): Ishihara H
  • Journal Title: Diabetology International Volume: - Issue: 3 Pages: 463-470
  • DOI: 10.1007/s13340-022-00576-z
  • Peer Reviewed / Open Access
• [Journal Article] Fibrosis-4 index is closely associated with arterial damage and future risk of coronary heart disease in type 2 diabetes (2022)
  • Author(s): Watanabe K, Takakubo N, Saigusa T, Nagasawa A, Yamana M, Ojima M, Kameda W, Susa S, Ishizawa K, Ishihara H.
  • Journal Title: Int J Hypertension Volume: 2022 Pages: 2760027-2760027
  • DOI: 10.1155/2022/2760027
  • Peer Reviewed / Open Access
• [Journal Article] Aortic arch calcification with pericardial fat mass detected on a single chest X-ray image is closely associated with the predictive variables of future cardiovascular disease (2022)
  • Author(s): Watanabe K, Hada Y, Ishii K, Nagaoka K, Takase K, Kameda W, Susa S, Saigusa T, Egashira F, Ishihara H, Ishizawa K.
  • Journal Title: Heart Vessels Volume: 37 Issue: 4 Pages: 654-664
  • DOI: 10.1007/s00380-021-01948-2
  • Peer Reviewed / Open Access
• [Journal Article] Using recombinase-mediated cassette exchange to engineer MIN6 insulin-secreting cells based on a newly identified safe harbor locus (2021)
  • Author(s): Furukawa A, Tanaka A, Yamaguchi S, Kosuda M, Yamana M, Nagasawa A, Kohno G, Ishihara H.
  • Journal Title: Journal of Diabetes Investigation Volume: 12 Issue: 12 Pages: 2129-2140
  • DOI: 10.1111/jdi.13646
  • Peer Reviewed / Open Access
• [Journal Article] Effects of GLP-1RA and SGLT2i, alone or in combination, on mouse models of type 2 diabetes representing different disease stages (2021)
  • Author(s): Koike M, Saito H, Kohno G, Takubo M, Watanabe K, Ishihara H.
  • Journal Title: International J Molecular Science Volume: 22 Issue: 21 Pages: 11463-11463
  • DOI: 10.3390/ijms222111463
  • Peer Reviewed / Open Access
• [Journal Article] Impact of Plasma Xanthine Oxidoreductase Activity on the Mechanisms of Distal Symmetric Polyneuropathy Development in Patients with Type 2 Diabetes (2021)
  • Author(s): Fujishiro Midori、Ishihara Hisamitsu、Ogawa Katsuhiko、Murase Takayo、Nakamura Takashi、Watanabe Kentaro、Sakoda Hideyuki、Ono Hiraku、Yamamotoya Takeshi、Nakatsu Yusuke、Asano Tomoichiro、Kushiyama Akifumi
  • Journal Title: Biomedicines Volume: 9 Issue: 8 Pages: 1052-1052
  • DOI: 10.3390/biomedicines9081052
  • Peer Reviewed / Open Access
• [Journal Article] Glucagon responses to glucose challenge in patients with idiopathic postprandial syndrome (2021)
  • Author(s): Kosuda M, Watanabe K, Koike M, Morikawa A, Saito H, Kohno G, Ishihara H.
  • Journal Title: J Nippon Medical School Volume: -
  • NAID: 130008087911
  • Peer Reviewed
• [Journal Article] Effect of adult weight gain on non-alcoholic fatty liver disease and its association with anthropometric parameters in the lean Japanese population (2020)
  • Author(s): Tanaka S, Fujishiro M, Watanabe K, Imatake K, Suzuki Y, Abe M, Ishihara H, Tani S.
  • Journal Title: Diagnostics Volume: 10 Issue: 11 Pages: 863-863
  • DOI: 10.3390/diagnostics10110863
  • Peer Reviewed / Open Access
• [Journal Article] A rare case of fulminant type 1 diabetes mellitus accompanied by both acute pancreatitis and myocarditis - case report (2020)
  • Author(s): Egashira F, Kawashima M, Morikawa A, Kosuda M, Ishihara H, Watanabe K.
  • Journal Title: BMC Endocr Disord Volume: 20 Issue: 1 Pages: 127-127
  • DOI: 10.1186/s12902-020-00607-3
  • Peer Reviewed / Open Access
• [Journal Article] Comparison of tofogliflozin versus glimepiride as the third oral agent added to metformin plus a dipeptidyl peptidase-4 inhibitor in Japanese patients with type 2 diabetes: A randomized, 24-week, open-label, controlled trial (STOP-OB) (2020)
  • Author(s): Kitazawa T, Seino H, Ohashi H, Inazawa T, Inoue M, Ai M, Fujishiro M, Kuroda H, Yamada M, Anai M, Ishihara H.
  • Journal Title: Diabetes Obes Metab Volume: 22 Issue: 9 Pages: 1659-1663
  • DOI: 10.1111/dom.14059
  • Peer Reviewed / Open Access
• [Journal Article] Seasonal variation in hydration status among community-dwelling elderly in Japan (2020)
  • Author(s): Tanaka S, Fujishiro M, Watanabe K, Imatake K, Suzuki Y, Abe M, Ishihara H, Tani S
  • Journal Title: Geriatr Gerontol Int Volume: 20 Issue: 10 Pages: 904-910
  • DOI: 10.1111/ggi.14010
  • Peer Reviewed
• [Presentation] 膵a細胞株aTC1およびb細胞株MIN6からなる偽膵島の形態とインスリン分泌応答 (2022)
  • Author(s): 盛川愛 斉藤一樹 河野玄太 田久保正洋 小須田南 石原寿光
  • Organizer: 第65回日本糖尿病学会年次学術集会
• [Presentation] グルコース応答性インスリン分泌におけるTCA回路酵素の役割の検討 (2022)
  • Author(s): 河野玄太 山名碧 小須田南 石原寿光
  • Organizer: 第65回日本糖尿病学会年次学術集会
• [Presentation] 時計遺伝子chrono (Ciart) 欠損マウスにおける糖代謝の解析 (2022)
  • Author(s): 斉藤一樹 山口賢 田久保正洋 内匠透 河野玄太 石原寿光
  • Organizer: 第65回日本糖尿病学会年次学術集会
• [Presentation] グルカゴン分泌制御機構研究の新展開：膵島の統合的理解を目指して (2022)
  • Author(s): 石原 寿光
  • Organizer: 第59回日本糖尿病学会関東甲信越地方会（WEB開催）
  • Invited
• [Presentation] 効率的に遺伝子発現修飾が可能なMIN6細胞の作製と応用 (2021)
  • Author(s): 石原 寿光
  • Organizer: 第64回日本糖尿病学会年次学術総会（WEB開催）
  • Invited
• [Presentation] 2型糖尿病モデルマウスを用いたGLP-1受容体作動薬およびSGLT2阻害薬の薬効メカニズムの解析 (2021)
  • Author(s): 小池 将夫、斎藤 一樹、田久保 正洋、渡邉 健太郎、石原 寿光
  • Organizer: 第64回日本糖尿病学会年次学術総会（WEB開催）
• [Presentation] Postprandial syndrome におけるグルカゴン分泌の解析 (2021)
  • Author(s): 小須田 南、渡邉 健太郎、小池 将夫、盛川 愛、斎藤 一樹、河野 玄太、石原 寿光
  • Organizer: 第64回日本糖尿病学会年次学術総会（WEB開催）
• [Presentation] 2型糖尿病の経過中に多脾症合併膵体尾部低形成症が判明した1例 (2021)
  • Author(s): 田久保正洋，小須田南，長澤瑛子，山名碧，江頭富士子，渡邉健太郎，今津博雄，石原寿光
  • Organizer: 第58回日本糖尿病学会関東甲信越地方会（WEB開催）
• [Presentation] ミトコンドリアピルビン酸輸送担体欠損インスリン分泌細胞株の樹立と解析 (2020)
  • Author(s): 西岡秀典，山名碧，長澤瑛子，小須田南，渡邉健太郎，石原寿光
  • Organizer: 第63回日本糖尿病学会年次学術集会（WEB開催）
• [Presentation] Roles of glycolysis distinct from mitochondrial metabolism for signal generation in Insulin secreting cells (2020)
  • Author(s): Ishihara H, Nishioka H, Yamana M, Nagasawa A, Kosuda M, Koike M, Kohno G, Saito H
  • Organizer: American Diabetes Association 80th Scientific Sessions
  • Int'l Joint Research
• [Patent(Industrial Property Rights)] 動物細胞の染色体上への外来遺伝子の挿入方法 (2020)
  • Inventor(s): 石原 寿光
  • Industrial Property Rights Holder: 石原 寿光
  • Industrial Property Rights Type: 特許
  • Filing Date: 2020