Mechanism and prevention strategy of inflammation and fibrosis caused by quantitative and qualitative transformation of organ lipids in inflammation cellular society

• Project Area: Preventive medicine through inflammation cellular sociology
• Project/Area Number: 17H06395
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Complex systems
• Research Institution: University of Tsukuba
• Principal Investigator: 島野 仁 筑波大学, 医学医療系, 教授 (20251241)
• Co-Investigator(Kenkyū-buntansha): 松坂 賢 筑波大学, 医学医療系, 教授 (70400679) ; 中川 嘉 筑波大学, 国際統合睡眠医科学研究機構, 准教授 (80361351)
• Project Period (FY): 2017-06-30 – 2022-03-31
• Project Status: Granted (Fiscal Year 2020)
• Budget Amount: ¥91,520,000 (Direct Cost: ¥70,400,000、Indirect Cost: ¥21,120,000); Fiscal Year 2021: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000); Fiscal Year 2020: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000); Fiscal Year 2019: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000); Fiscal Year 2018: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000); Fiscal Year 2017: ¥21,320,000 (Direct Cost: ¥16,400,000、Indirect Cost: ¥4,920,000)
• Keywords: 脂質 / 炎症 / 線維化 / 糖尿病 / 異質 / 膵臓ラ氏島 / NASH / 未病 / １細胞解析 / 生活習慣病 / 慢性炎症 / 非アルコール性脂肪肝 / 脂質代謝 / CREBH / ChREBP / 動脈硬化 / Elovl6 / SREBP / 肥満 / 発現制御

Outline of Annual Research Achievements

２型糖尿病発症における炎症、線維化と病態の進行の網羅的に把握して新規のメカニズムを知ることが本研究の主題の一つである。　肥満２型糖尿病モデルdbdbマウスの正常対照、糖尿病発症前（６週齢）、発症後（１０週齢）の３病態における膵臓ラ氏島を単離して、昨年度まで条件検討の準備を行なっていたsingle cell transcriptome analysis を施行し、　病態に関わる細胞クラスターやマーカーないし責任遺伝子の探索をおこなった。　本年度は、特に病態の本体であるベータ細胞に注目して、正常から未病発症にわたってベータ細胞を５つのクラスターに分離できることを初めて示すことができた。　ベータ細胞で糖尿病態に関与が知られている因子の予想通りの変化が確認されるとともに、未病期にで特異的に上昇する新規の分子が発見され優れた病初期マーカーの可能性を詳細に検討している。
肝臓の脂質代謝と炎症、線維化の関連を探索する目的で実施している新規プロテアーゼ（R4）による脂質種特異的なSREBP-1活性化制御機構の解析では、in vitro実験の結果、 R4が飽和脂肪酸によるSREBP-1の活性化を亢進し、不飽和脂肪酸によるSREBP-1活性化抑制に働くことを明らかにした。また、マウス肝臓を用いたin vivo実験において、不飽和脂肪酸EPAによるSREBP-1 活性化抑制がR4ノックアウトマウスでは減弱することから、EPAによるSREBP-1抑制作用にR4が関わっていることを明らかにした。　さらに脂質分解、ストレス応答転写CREBHのKOマウスの高脂肪食負荷実験の肝臓では、著明に炎症、線維化が悪化し、数ヶ月後には多発性に腫瘍が発生した。今後、NASH-LC-HCCのモデルとして確立後SCTを検討する。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

本研究課題の技術的バックボーンであるsingle cell transcriptome analysisは、　組織からの細胞の１つ１つの単離が鍵であり、膵臓ラ氏島のサンプル抽出に当初苦労したが、条件や酵素の改良の結果、非常２つの質の高いサンプルとデータを得られた。　その困難さから、発生期のラ氏島やヒトのサンプルでの報告にかぎられており、新規病初期マーカーの発見を軸に、２型糖尿病モデルにおける初のSCT報告を目指す。
肝臓のSREBP-1の脂肪酸種特異的切断酵素の発見はほぼ解析がすみ投稿を予定している。

Strategy for Future Research Activity

膵臓実績の概要に記したように、２型糖尿病の病態にあたらしい解釈を与えるストーリーが見えてきており今後解析する。 また５つのクラスターを病態進行で俯瞰して、遺伝子群パスウェイを検討するとERストレスやER-golgiのソーティングに関する遺伝子が抽出され、炎症、線維化に先行する病態が炙り出された。今後、詳細に解析するとともにベータ細胞以外のクラスターとの相互作用を検討していく。　ベータ細胞のみならず、アルファ細胞、デルタ細胞、PP細胞、内皮細胞、単球マクロファージ系、ストローマ系、ductal, 腺房細胞など多くの細胞種がクラスターとして分類されており、pseudtime 解析、interactome解析を実施予定して、病態との関連を明らかにしていく。
肝臓のSREBP-1の新規特異的切断酵素は、膜タンパクであるが膜の流動性やストレス応答に関わっている様で、今後炎症特に線維化との関連を明らかにしていく。

Research Products

• [Journal Article] New perspective on type?2 diabetes, dyslipidemia and non‐alcoholic fatty liver disease (2020)
  • Author(s): Matsuzaka Takashi、Shimano Hitoshi
  • Journal Title: J Diabetes Investig. Volume: 11 Pages: 532-534
  • DOI: 10.1111/jdi.13258
  • Peer Reviewed / Open Access
• [Journal Article] CREBH Improves Diet-Induced Obesity, Insulin Resistance, and Metabolic Disturbances by FGF21-Dependent and FGF21-Independent Mechanisms (2020)
  • Author(s): Satoh A, Han S, Araki M, Nakagawa Y, Ohno H, Mizunoe Y, Kumagai K, Murayama Y, Osaki Y, Iwasaki H, Sekiya M, Konishi M, Itoh N, Matsuzaka T, Sone H, and Shimano H.
  • Journal Title: iScience Volume: 23 Issue: 3 Pages: 100930-100930
  • DOI: 10.1016/j.isci.2020.100930
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Hepatocyte ELOVL Fatty Acid Elongase 6 Determines Ceramide Acyl-Chain Length and Hepatic Insulin Sensitivity in Mice (2020)
  • Author(s): Matsuzaka T, Kuba M, Koyasu S, Takeuchi Y, Yahagi N, Miyamoto T, Sekiya M, Nakagawa Y, Ema M, Takahashi S, Tokiwa H, Shimano H.
  • Journal Title: Hepatology Volume: - Pages: 1609-1625
  • DOI: 10.1002/hep.30953
  • Description: in press
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Handgrip strength predicts new prediabetes cases among adults: A prospective cohort study (2020)
  • Author(s): Manda Chrispin Mahala、Hokimoto Takahiro、Okura Tomohiro、Isoda Hiroko、Shimano Hitoshi、Wagatsuma Yukiko
  • Journal Title: Preventive Medicine Reports Volume: 17 Pages: 101056-101056
  • DOI: 10.1016/j.pmedr.2020.101056
  • Peer Reviewed / Open Access
• [Journal Article] Cathepsin B overexpression induces degradation of perilipin 1 to cause lipid metabolism dysfunction in adipocytes (2020)
  • Author(s): Mizunoe Yuhei、Kobayashi Masaki、Hoshino Shunsuke、Tagawa Ryoma、Itagawa Rei、Hoshino Ayana、Okita Naoyuki、Sudo Yuka、Nakagawa Yoshimi、Shimano Hitoshi、Higami Yoshikazu
  • Journal Title: Sci Rep. Volume: 10 Pages: 634-634
  • DOI: 10.1038/s41598-020-57428-6
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Elucidation of Molecular Mechanism of a Selective PPARα Modulator, Pemafibrate, through Combinational Approaches of X-ray Crystallography, Thermodynamic Analysis, and First-Principle Calculations (2020)
  • Author(s): Kawasaki Mayu、Kambe Akira、Yamamoto Yuta、Arulmozhiraja Sundaram、Ito Sohei、Nakagawa Yoshimi、Tokiwa Hiroaki、Nakano Shogo、Shimano Hitoshi
  • Journal Title: Int J Mol Sci. Volume: 21 Pages: 361-361
  • DOI: 10.3390/ijms21010361
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Transcriptional co-repressor CtBP2 orchestrates epithelial-mesenchymal transition through a novel transcriptional holocomplex with OCT1 (2020)
  • Author(s): Ma Yang、Sekiya Motohiro、Kainoh Kenta、Matsuda Takaaki、Iwasaki Hitoshi、Osaki Yoshinori、Sugano Yoko、Suzuki Hiroaki、Takeuchi Yoshinori、Miyamoto Takafumi、Yahagi Naoya、Nakagawa Yoshimi、Matsuzaka Takashi、Shimano Hitoshi
  • Journal Title: Biochem Biophys Res Commun. Volume: 523 Pages: 354-360
  • DOI: 10.1016/j.bbrc.2019.12.070
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Association between Lysosomal Dysfunction and Obesity-Related Pathology: A Key Knowledge to Prevent Metabolic Syndrome (2019)
  • Author(s): Mizunoe Yuhei、Kobayashi Masaki、Tagawa Ryoma、Nakagawa Yoshimi、Shimano Hitoshi、Higami Yoshikazu
  • Journal Title: Int J Mol Sci. Volume: 20 Pages: 3688-3688
  • DOI: 10.3390/ijms20153688
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The selective peroxisome proliferator-activated receptor alpha modulator (SPPARMα) paradigm: conceptual framework and therapeutic potential (2019)
  • Author(s): Fruchart Jean-Charles、Santos Raul D.Ponte-Negretti Carlos I、Pradhan Aruna、Ray Kausik K.、Reiner ?eljko、Ridker Paul M.、Ruscica Massimiliano、Sadikot Shaukat、Shimano Hitoshi. et al
  • Journal Title: Cardiovasc Diabetol. Volume: 18 Pages: 71-71
  • DOI: 10.1186/s12933-019-0864-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Rho‐associated, coiled‐coil‐containing protein kinase?1 as a new player in the regulation of hepatic lipogenesis (2019)
  • Author(s): Miyamoto Takafumi、Matsuzaka Takashi、Shimano Hitoshi
  • Journal Title: J Diabetes Investig. Volume: 10 Pages: 1165-1167
  • DOI: 10.1111/jdi.13051
  • Peer Reviewed / Open Access
• [Journal Article] Octacosanol and policosanol prevent high-fat diet-induced obesity and metabolic disorders by activating brown adipose tissue and improving liver metabolism (2019)
  • Author(s): Sharma Rahu, Takafumi Miyamoto, et al.
  • Journal Title: Sci Rep Volume: 9 Pages: 1-12
  • DOI: 10.1038/s41598-019-41631-1
  • NAID: 120006714297
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Transcriptome network analysis identifies protective role of the LXR/SREBP-1c axis in murine pulmonary fibrosis. (2019)
  • Author(s): Shichino S, Ueha S, Hashimoto S, Otsuji M, Abe J, Tsukui T, Deshimaru S, Nakajima T, Kosugi-Kanaya M, Shand FH, Inagaki Y, Shimano H, Matsushima K.
  • Journal Title: JCI Insight Volume: 4(1) Pages: 122163-122163
  • DOI: 10.1172/jci.insight.122163
  • Peer Reviewed / Open Access
• [Journal Article] Elovl6 regulates mechanical damage-induced keratinocyte death and skin inflammation. (2018)
  • Author(s): Nakamura Y, Matsuzaka T, Tahara-Hanaoka S, Shibuya K, Shimano H, Nakahashi-Oda C, Shibuya A.
  • Journal Title: Cell Death and Diseases Volume: 9(12) Pages: 1181-1181
  • DOI: 10.1038/s41419-018-1226-1
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The Peroxisome Proliferator-Activated Receptor α (PPARα) Agonist Pemafibrate Protects against Diet-Induced Obesity in Mice (2018)
  • Author(s): Araki Masaya、Nakagawa Yoshimi、Oishi Asayo、Han Song-iee、Wang Yunong、Kumagai Kae、Ohno Hiroshi、Mizunoe Yuhei、Iwasaki Hitoshi、Sekiya Motohiro、Matsuzaka Takashi、Shimano Hitoshi
  • Journal Title: International journal of molecular sciences Volume: 19 Pages: 2148-2148
  • DOI: 10.3390/ijms19072148
  • NAID: 120006531296
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Transgenic Mice Overexpressing SREBP-1a in Male ob/ob Mice Exhibit Lipodystrophy and Exacerbate Insulin Resistance. (2018)
  • Author(s): Ohno H, Matsuzaka T, Tang N, Sharma R, Motomura K, Shimura T, Satoh A, Han SI, Takeuchi Y, Aita Y, Iwasaki H, Yatoh S, Suzuki H, Sekiya M, Nakagawa Y, Sone H, Yahagi N, Yamada N, Higami Y, Shimano H.
  • Journal Title: Endocrinology Volume: 159(6) Pages: 2308-2323
  • DOI: 10.1210/en.2017-03179
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Molecular association model of PPARα and its new specific and efficient ligand, pemafibrate: Structural basis for SPPARMα (2018)
  • Author(s): Yamamoto Yuta、Takei Kenta、Arulmozhiraja Sundaram、Sladek Vladimir、Matsuo Naoya、Han Song-iee、Matsuzaka Takashi、Sekiya Motohiro、Tokiwa Takaki、Shoji Mitsuo、Shigeta Yasuteru、Nakagawa Yoshimi、Tokiwa Hiroaki、Shimano Hitoshi
  • Journal Title: Biochem Biophys Res Commun Volume: 499 Pages: 239-245
  • DOI: 10.1016/j.bbrc.2018.03.135
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Saturated Fatty Acids Undergo Intracellular Crystallization and Activate the NLRP3 Inflammasome in MacrophagesHighlights (2018)
  • Author(s): Karasawa Tadayoshi、Kawashima Akira、Usui-Kawanishi Fumitake、Watanabe Sachiko、Kimura Hiroaki、Kamata Ryo、Shirasuna Koumei、Koyama Yutaro、Sato-Tomita Ayana、Matsuzaka Takashi、Tomoda Hiroshi、Park Sam-Yong、Shibayama Naoya、Shimano Hitoshi、Kasahara Tadashi、Takahashi Masafumi
  • Journal Title: Arterioscler Thromb Vasc Biol. Volume: 38 Pages: 744-756
  • DOI: 10.1161/atvbaha.117.310581
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Epigenetic modulation of Fgf21 in the perinatal mouse liver ameliorates diet-induced obesity in adulthood (2018)
  • Author(s): X Yuan, K Tsujimoto, K Hashimoto, K Kawahori, N Hanzawa, M Hamaguchi, T Seki, M Nawa, T Ehara, Y Kitamura, I Hatada, M Konishi, N Itoh, Y Nakagawa, H Shimano, T Takai-Igarashi, Y Kamei, Y Ogawa.
  • Journal Title: Nat. Commun. Volume: 9 Pages: 636-636
  • DOI: 10.1038/s41467-018-03038-w
  • NAID: 120006454799
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Epigenetic modulation of Fgf21 in the perinatal mouse liver ameliorates diet-induced obesity in adulthood. (2018)
  • Author(s): Yuan X, Tsujimoto K, Hashimoto K, Kawahori K, Hanzawa N, Hamaguchi M, Seki T, Nawa M, Ehara T, Kitamura Y, Hatada I, Konishi M, Itoh N, Nakagawa Y, Shimano H, Takai-Igarashi T, Kamei Y, Ogawa Y.
  • Journal Title: Nat Commun. Volume: 9(1) Pages: 636-636
  • NAID: 120006454799
  • Peer Reviewed / Open Access
• [Journal Article] A candidate functional SNP rs7074440 in TCF7L2 alters gene expression through C-FOS in hepatocytes. (2018)
  • Author(s): Piao X, Yahagi N, Takeuchi Y, Aita Y, Murayama Y, Sawada Y, Shikama A, Masuda Y, Nishi-Tatsumi M, Kubota M, Izumida Y, Sekiya M, Matsuzaka T, Nakagawa Y, Sugano Y, Iwasaki H, Kobayashi K, Yatoh S, Suzuki H, Yagyu H, Kawakami Y, Shimano H.
  • Journal Title: FEBS Letters Volume: 592(3) Pages: 422-433
  • DOI: 10.1002/1873-3468.12975
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Novel non-alcoholic steatohepatitis model with histopathological and insulin-resistant features. (2018)
  • Author(s): Owada Y, Tamura T, Tanoi T, Ozawa Y, Shimizu Y, Hisakura K, Matsuzaka T, Shimano H, Nakano N, Sakashita S, Matsukawa T, Isoda H, Ohkohchi N.
  • Journal Title: Pathol Int. Volume: 68(1) Pages: 12-22
  • DOI: 10.1111/pin.12612
  • NAID: 120006399145
  • Peer Reviewed
• [Journal Article] Critical role of CREBH-mediated induction of transforming growth factor β2 by hepatitis C virus infection in fibrogenic responses in hepatic stellate cells (2018)
  • Author(s): Chida Takeshi、Ito Masahiko、Nakashima Kenji、Kanegae Yumi、Aoshima Takuya、Takabayashi Shuji、Kawata Kazuhito、Nakagawa Yoshimi、Yamamoto Masahiro、Shimano Hitoshi、Matsuura Tomokazu、Kobayashi Yoshimasa、Suda Takafumi、Suzuki Tetsuro
  • Journal Title: Hepatology. Volume: 66 Pages: 1430-1443
  • DOI: 10.1002/hep.29319
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Mitochondrial intermediate peptidase is a novel regulator of sirtuin-3 activation by caloric restriction (2017)
  • Author(s): Kobayashi Masaki、Takeda Kanae、Narita Takumi、Nagai Keita、Okita Naoyuki、Sudo Yuka、Miura Yuri、Tsumoto Hiroki、Nakagawa Yoshimi、Shimano Hitoshi、Higami Yoshikazu
  • Journal Title: FEBS Lett Volume: 591 Pages: 4067-4073
  • DOI: 10.1002/1873-3468.12914
  • NAID: 120006401862
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Effect of sodium-glucose cotransporter 2 (SGLT2) inhibition on weight loss is partly mediated by liver-brain-adipose neurocircuitry (2017)
  • Author(s): Sawada Yoshikazu、Izumida Yoshihiko、Takeuchi Yoshinori、Aita Yuichi、Wada Nobuhiro、Li EnXu、Murayama Yuki、Piao Xianying、Shikama Akito、Masuda Yukari、Nishi-Tatsumi Makiko、Kubota Midori、Sekiya Motohiro、Matsuzaka Takashi、Nakagawa Yoshimi、Shimano Hitoshi、Yahagi Naoya
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 493 Pages: 40-45
  • DOI: 10.1016/j.bbrc.2017.09.081
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] 12. SREBP-regulated lipid metabolism: convergent physiology - divergent pathophysiology. (2017)
  • Author(s): Shimano H, Sato R.
  • Journal Title: Nat Rev Endocrinol. Volume: 13(12) Pages: 710-730
  • DOI: 10.1038/nrendo.2017.91
  • Peer Reviewed
• [Journal Article] Elovl6 Deficiency Improves Glycemic Control in Diabetic db / db Mice by Expanding β-Cell Mass and Increasing Insulin Secretory Capacity (2017)
  • Author(s): Zhao Hui、Matsuzaka Takashi、Nakano Yuta、Motomura Kaori、Tang Nie、Yokoo Tomotaka、Okajima Yuka、Han Song-iee、Takeuchi Yoshinori、Aita Yuichi、Iwasaki Hitoshi、Yatoh Shigeru、Suzuki Hiroaki、Sekiya Motohiro、Yahagi Naoya、Nakagawa Yoshimi、Sone Hirohito、Yamada Nobuhiro、Shimano Hitoshi
  • Journal Title: Diabetes. Volume: 66 Pages: 1833-1846
  • DOI: 10.2337/db16-1277
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 心血管疾患のリスク病態としてのNAFLD/NASH（合同シンポジウム） (2019)
  • Author(s): 島野仁
  • Organizer: 第51回日本動脈硬化学会総会・学術集会
  • Invited
• [Presentation] 脂質の量と質の変容がもたらす臓器病態と栄養戦略 (2019)
  • Author(s): 島野仁
  • Organizer: 第73回日本栄養・食糧学会大会
  • Invited
• [Presentation] A novel aspect of fatty acid chain length in lipotoxicity: Lessons from Elovl6 KO mice (2019)
  • Author(s): Hitoshi Shimano
  • Organizer: 60th International Conference on the Bioscience of Lipids
  • Int'l Joint Research / Invited
• [Presentation] A metabolite sensor CtBP2 integrates metabolic flux to orchestrate glucose and lipid homeostasis in liver (2019)
  • Author(s): Motohiro Sekiya, Hitoshi Shimano, Gokhan Hotamisligil
  • Organizer: 60th International Conference on the Bioscience of Lipids
  • Int'l Joint Research / Invited
• [Presentation] CREB3L3欠損による動脈硬化発症の分子メカニズム (2017)
  • Author(s): 中川 嘉、関谷 元博、松坂 賢、島野 仁
  • Organizer: 第32回 日本糖尿病合併症学会
• [Remarks] 筑波大学 医学医療系 内分泌代謝・糖尿病内科
  • URL: https://www.u-tsukuba-endocrinology.jp/
• [Remarks] 筑波大学 医学医療系 内分泌代謝・糖尿病代謝内科
  • URL: http://www.u-tsukuba-endocrinology.jp/index.html
• [Patent(Industrial Property Rights)] 生細胞内論理ゲートによるオルガネラの形態操作法 (2019)
  • Inventor(s): 宮本崇史、松坂賢、島野仁
  • Industrial Property Rights Holder: 宮本崇史、松坂賢、島野仁
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2019-211266
  • Filing Date: 2019