Integrated analysis of microRNAs that regulate cartilage damage and its functional analysis in chondrocytes

• Project/Area Number: 26293338
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Orthopaedic surgery
• Research Institution: Okayama University
• Principal Investigator: HIROHATA SATOSHI 岡山大学, 保健学研究科, 教授 (90332791)
• Co-Investigator(Kenkyū-buntansha): 伊藤 佐智夫 岡山大学, 医歯(薬)学総合研究科, 助教 (30335624) ; 成瀬 恵治 岡山大学, 医歯(薬)学総合研究科, 教授 (40252233) ; 西田 圭一郎 岡山大学, 医歯(薬)学総合研究科, 准教授 (80284058) ; 大月 孝志 岡山大学, 保健学研究科, 非常勤研究員 (10534802)
• Research Collaborator: NINOMIYA Yoshifumi 岡山大学, 医歯(薬)学総合研究科, 教授 (70126241)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥16,250,000 (Direct Cost: ¥12,500,000、Indirect Cost: ¥3,750,000); Fiscal Year 2016: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2015: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2014: ¥8,450,000 (Direct Cost: ¥6,500,000、Indirect Cost: ¥1,950,000)
• Keywords: 遺伝子 / アグリカナーゼ / 関節病学 / 細胞外マトリックス

Outline of Final Research Achievements

Aggrecan is a major proteoglycan in cartilage and is degraded by aggrecanase. Aggrecanases plays a role in the early stage of osteoarthritis. This research aimed the integrated analysis of microRNAs whose targets are the aggrecanase mRNAs and extracellular matrix proteins. We try to examine how aggrecanases are regulated by microRNAs.
In this study, we first stimulated chondrocytic cells by cytokine as well as mechanical stress. We then compared the change of microRNA expression between two different stimulations. By using several database, we identified the target gene for microRNAs of interest. Then we added another stimulation condition and selected the microRNA of interest. We next examined the effect of inhibitor of microRNA and confirmed that inhibitor abolished the effect of microRNA. Finally, we confirmed that the target of microRNA is actually proteases that induced in osteoarthritis. In conclusion, we identified the novel microRNA that may play a role in osteoarthritis.

Research Products

• [Journal Article] Early initiation of eicosapentaenoic acid and statin treatment is associated with better clinical outcomes than statin alone in patients with acute coronary syndromes: 1-year outcomes of a randomized controlled study. (2017)
  • Author(s): Nosaka K, Miyoshi T, Iwamoto M, Kajiya M, Okawa K, Tsukuda S, Yokohama F, Sogo M, Nishibe T, Matsuo N, Hirohata S, Ito H, Doi M
  • Journal Title: Int J Cardiol. Volume: 228 Pages: 173-179
  • DOI: 10.1016/j.ijcard.2016.11.105
  • Peer Reviewed / Open Access
• [Journal Article] A rapid and precise method for measuring plasma apoE-rich HDL using polyethylene glycol and cation-exchange chromatography: a pilot study on the clinical significance of apoE-rich HDL measurements. (2017)
  • Author(s): Ikeda T, Shinohata R, Murakami M, Hina K, Kamikawa S, Hirohata S, Kusachi S, Tamura A, Usui S.
  • Journal Title: Clin Chim Acta. Volume: 465 Pages: 112-118
  • DOI: 10.1016/j.cca.2016.12.016
  • Peer Reviewed
• [Journal Article] Loss of ADAMTS4 reduces high fat diet-induced atherosclerosis and enhances plaque stability in ApoE(-/-) mice. (2016)
  • Author(s): Kumar S, Chen M, Li Y, Wong FH, Thiam CW, Hossain MZ, Poh KK, Hirohata S, Ogawa H, Angeli V, Ge R.
  • Journal Title: Sci Rep. Volume: 6 Issue: 1 Pages: 31130-31130
  • DOI: 10.1038/srep31130
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] MMP-13 is constitutively produced in human chondrocytes and co-endocytosed with ADAMTS-5 and TIMP-3 by the endocytic receptor LRP1 (2016)
  • Author(s): Yamamoto K, Okano H, Miyagawa W, Visse R, Shitomi Y, Santamaria S, Dudhia J, Troeberg L, Strickland DK, Hirohata S, Nagase H
  • Journal Title: Matrix Biol Volume: S0945-053X(16) Pages: 30031-30032
  • DOI: 10.1016/j.matbio.2016.03.007
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Subclavian steal syndrome: a case report and review of advances in diagnostic and treatment approaches. (2016)
  • Author(s): Komatsubara I, Kondo J, Akiyama M, Takeuchi H, Nogami K, Usui S, Hirohata S, Kusachi S
  • Journal Title: Cardiovasc Revasc Med Volume: 17(1) Issue: 1 Pages: 54-58
  • DOI: 10.1016/j.carrev.2015.11.001
  • Peer Reviewed
• [Journal Article] Eosinophil Cationic Protein Shows Survival Effect on H9c2 Cardiac Myoblast Cells with Enhanced Phosphorylation of ERK and Akt/GSK-3β under Oxidative Stress. (2015)
  • Author(s): Ishii H, Kamikawa S, Hirohata S, Mizutani A, Abe K, Seno M, Oohashi T, Ninomiya Y
  • Journal Title: Acta Med Okayama. Volume: 69（3） Pages: 145-153
  • NAID: 120005621908
  • Peer Reviewed
• [Journal Article] Interleukin-1β induced nuclear factor-κB binds to a disintegrin-like and metalloproteinase with thrombospondin type 1 motif 9 promoter in human chondrosarcoma cells. (2015)
  • Author(s): Altuntas A, Halacli SO, Cakmak O, Erden G, Akyol S, Ugurcu V, Hirohata S, Demircan K.
  • Journal Title: Mol Med Rep. Volume: in press Issue: 1 Pages: 595-600
  • DOI: 10.3892/mmr.2015.3444
  • Peer Reviewed
• [Journal Article] Low serum level of secreted frizzled-related protein 5, an anti-inflammatory adipokine, is associated with coronary artery disease. (2014)
  • Author(s): Miyoshi T, Doi M, Usui S, Iwamoto M, Kajiya M, Takeda K, Nosaka K, Nakayama R, Okawa K, Takagi W, Nakamura K, Hirohata S, Ito H
  • Journal Title: Atherosclerosis Volume: 233（2） Pages: 454-459
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] ADAMTS1 inhibits lymphangiogenesis by attenuating phosphorylation of the lymphatic endothelial cell-specific VEGF receptor. (2014)
  • Author(s): Inagaki J, Takahashi K, Ogawa H, Asano K, Hatipoglu OF, Zeynel Cilek M, Obika M, Ohtsuki T, Hofmann M, Kusachi S, Ninomiya Y, Hirohata S
  • Journal Title: Exp Cell Res. Volume: 323(2) Pages: 263-275
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] ADAMTS4 and ADAMTS5 Knockout Mice Are Protected from Versican but Not Aggrecan or Brevican Proteolysis during Spinal Cord Injury. (2014)
  • Author(s): Demircan K, Topcu V, Takigawa T, Akyol S, Yonezawa T, Ozturk G, Ugurcu V, Hasgul R, Yigitoglu MR, Akyol O, McCulloch DR, Hirohata S.
  • Journal Title: Biomed Res Int. Volume: 2014 Pages: 693746-693746
  • DOI: 10.1155/2014/693746
  • Peer Reviewed / Open Access
• [Journal Article] Early eicosapentaenoic acid treatment after percutaneous coronary intervention reduces acute inflammatory responses and ventricular arrhythmias in patients with acute myocardial infarction: A randomized, controlled study. (2014)
  • Author(s): Doi M, Nosaka K, Miyoshi T, Iwamoto M, Kajiya M, Okawa K, Nakayama R, Takagi W, Takeda K, Hirohata S, Ito H
  • Journal Title: Int J Cardiol. Volume: 176(3) Issue: 3 Pages: 577-582
  • DOI: 10.1016/j.ijcard.2014.08.055
  • Peer Reviewed
• [Presentation] Molecular and biochemical analysis of HYBID (HYaluronan-Binding protein Involved in hyaluronan Depolymerization) gene expression (2017)
  • Author(s): Ohtsuki T, Yoshida H, Shinaoka A, Kumagishi-Shinaoka K, Cilek MZ, Hatipoglu OF, Inagaki J, Nishida K,Okada Y, Hirohata S.
  • Organizer: 第30回日本軟骨代謝学会
  • Place of Presentation: 京都市
  • Year and Date: 2017-03-03
• [Presentation] Long forced inspiratory time is associated with high transpulmonary pressure in patients with acute respiratory failure. (2016)
  • Author(s): Hayashi K, Hirayama T, Yumoto T, Tsukahara K, Iida A, Nosaka N, Sato K, Ugawa T, Nakao A, Hirohata S.
  • Organizer: 欧州集中治療学会
  • Place of Presentation: イタリア ミラノ
  • Year and Date: 2016-10-01
  • Int'l Joint Research
• [Presentation] LDL particle size associates with apoE-containing HDL in patients who undergo coronary computed tomographic angiography. (2016)
  • Author(s): Shinohata R, Ikeda T, Murakami M, Hina K, Kamikawa S, Hirohata S, Kusachi S, Nakajima K, Ikeda S, Usui S.
  • Organizer: 米国臨床化学会(AACC)年次集会
  • Place of Presentation: 米国フィラデルフィア
  • Year and Date: 2016-07-31
  • Int'l Joint Research
• [Presentation] 変形性関節症へのヒアルロン酸投与、いつ投与すべきか？～動物モデルからの知見～ (2016)
  • Author(s): 大月孝志、品岡 玲、熊岸香苗、河野真優美、篠原真歩、浅野恵一、稲垣純子、大橋俊孝、西田圭一郎、廣畑 聡
  • Organizer: 第48回日本結合組織学会学術大会
  • Place of Presentation: 長崎市
  • Year and Date: 2016-06-24
• [Presentation] Distribution of versican and its degradation by ADAMTS protease in the tumor microenvironment. (2016)
  • Author(s): Satoshi Hirohata, Keiichi Asano, Junko Inagaki, Takashi Ohtsuki, Nao Yamanokuchi, Shogo Watanabe, Toshitaka Oohashi
  • Organizer: KSBMB International Conference 2016
  • Place of Presentation: 韓国ソウル市
  • Year and Date: 2016-05-18
  • Int'l Joint Research / Invited
• [Presentation] Distribution of versican and its degradation by ADAMTS protease in the tumor microenvironment (2016)
  • Author(s): Satoshi Hirohata, Keiichi Asano, Junko Inagaki, Takashi Ohtsuki, Nao Yamanokuchi, Shogo Watanabe, Toshitaka Oohashi
  • Organizer: KSBMB International Conference 2016
  • Place of Presentation: ソウル
  • Year and Date: 2016-05-18
  • Int'l Joint Research / Invited
• [Presentation] The diverse function of a disintegrin and metalloproteinase with thrombospondin motif-1 (ADAMTS1) (2016)
  • Author(s): Satoshi Hirohata
  • Organizer: TIDRC International Symposium -Inflammation and Tumor microenvironment-
  • Place of Presentation: 韓国ソウル市
  • Year and Date: 2016-05-17
  • Int'l Joint Research / Invited
• [Presentation] The diverse function of a disintegrin and metalloproteinase with thrombospondin motif-1 (ADAMTS1) (2016)
  • Author(s): Satoshi Hirohata
  • Organizer: TIDRC International Symposium -Inflammation and Tumor microenvironment-
  • Place of Presentation: ソウル
  • Year and Date: 2016-05-17
  • Int'l Joint Research / Invited
• [Presentation] 細胞外マトリックス分解酵素ADAMTS1の多様性 (2016)
  • Author(s): 廣畑 聡、大月孝志、稲垣純子
  • Organizer: 日本薬学会第136年会
  • Place of Presentation: 横浜市
  • Year and Date: 2016-03-27
  • Invited
• [Presentation] The Impact of Dyslipidemia and MDA-LDL on Coronary Slow Flow Phenomenon in Patients with Unstable Angina Pectoris (2016)
  • Author(s): Naofumi Amioka, Tatsuya Yamaji, Naoaki Matsuo, Kenzo Kagawa, Yukari Nakano, Daisuke Yamada, Nobuhiko Ohnishi, Atsushi Takaishi, Masayuki Ueeda, Satoshi Hirohata, Hiroshi Itoh
  • Organizer: 第８０回日本循環器学会学術集会
  • Place of Presentation: 仙台市
  • Year and Date: 2016-03-18
• [Presentation] ラット変形性関節症モデルへのヒアルロン酸投与による軟骨保護効果の解析 (2016)
  • Author(s): 大月孝志、品岡玲、熊岸加苗、佐藤美來、金道幸子、長谷川みさ、海原弘貴、浅野恵一、障子友理、稲垣純子、大橋俊孝、西田圭一郎、廣畑 聡
  • Organizer: 第29回日本軟骨代謝学会
  • Place of Presentation: 広島市
  • Year and Date: 2016-02-19
• [Presentation] Association of Extracellular Matrix Cleavage and Tumor Vasculature Formation (2016)
  • Author(s): Keiichi Asano, Junko Inagaki, Yuri Shoji, Takashi Ohtsuki, Toshitaka Oohashi and Satoshi Hirohata
  • Organizer: 第９回高度医療都市を創出する未来技術国際シンポジウム
  • Place of Presentation: 岡山市
  • Year and Date: 2016-02-05
  • Int'l Joint Research
• [Presentation] 伸展刺激の強度による細胞外マトリックスタンパクおよびマトリックス分解酵素発現への影響 (2015)
  • Author(s): 大月孝志、金道幸子、長谷川みさ、稲垣純子、浅野恵一、 障子友理、熊岸加苗、西田圭一郎、大橋俊孝、廣畑 聡
  • Organizer: BMB 2015
  • Place of Presentation: 神戸市
  • Year and Date: 2015-12-01
• [Presentation] ADAMTS1 null mice demonstrated omphalocele phenotype (2015)
  • Author(s): Satoshi Hirohata, Omer Faruk Hatipoglu, Eriko Kusunoki, Takashi Ohtsuki, Junko Inagaki, Shozo Kusachi, Yoshifumi Ninomiya.
  • Organizer: ゴードンカンファレンス matrix metalloproteinase
  • Place of Presentation: 米国・メイン州
  • Year and Date: 2015-08-02
  • Int'l Joint Research
• [Presentation] 変形性膝関節症の治療効果に関する生化学的解析～滑液内軟骨分解活性の検討～ (2015)
  • Author(s): 熊岸 加苗，大月 孝志，廣畑 聡, 河村 顕治， 大塚 愛二，坂田賢一郎，坂田 俊輔
  • Organizer: 第２８回日本臨床整形外科学会学術集会
  • Place of Presentation: 下関市
  • Year and Date: 2015-07-19
• [Presentation] HYALURONAN REDUCED EXTRACELLULAR MATRIX DEGRADATION BOTH IN VITRO AND IN VIVO (2015)
  • Author(s): Takashi Ohtsuki, Yuri Shoji, Keiichi Asano, Aya Hirata, Teruyuki Kawadi, Junko Ingaki, Kanae Kumagishi, Keiichiro Nishida, Toshitaka Oohashi, Satoshi Hirohata.
  • Organizer: HA2015
  • Place of Presentation: フィレンツェ
  • Year and Date: 2015-06-07 – 2015-06-11
• [Presentation] HYALURONAN TREATMENT REDUCED MATRIX DEGRADATION BOTH IN VITRO AND IN VIVO RAT OSTEOARTHRITIS MODEL (2015)
  • Author(s): Yuri Shoji, Keiichi Asano, Aya Hirata, Teruyuki Kawadi, Junko Ingaki, Kanae Kumagishi, Keiichiro Nishida, Toshitaka Oohashi, Satoshi Hirohata.
  • Organizer: 10th International conference HYALURONAN 2015
  • Place of Presentation: イタリア・フィレンツェ
  • Year and Date: 2015-06-07
  • Int'l Joint Research
• [Presentation] Perinatal lethality in ADAMTS1 deficient mouse (2014)
  • Author(s): Satoshi Hirohata, Omer Faruk Hatipoglu, Eriko Kusunoki, Takashi Ohtsuki, Junko Inagaki, Keiichi Asano, Yuri Shoji, Teruyuki Kawadi, Aya Hirata, Yoshifumi Ninomiya
  • Organizer: American Society for Matrix Biology Biennial Meeting
  • Place of Presentation: 米国 クリーブランド
  • Year and Date: 2014-10-12 – 2014-10-15
• [Presentation] ADAMTS cleaved versican in tumor angiogenic area (2014)
  • Author(s): Keiichi Asano, Junko Inagaki, Matthias Hofmann, Teruyuki Kawadi, Aya Hirata, Yuri Shoji, Takashi Ohtsuki, Yoshifumi Ninomiya, Satoshi Hirohata
  • Organizer: American Society for Matrix Biology Biennial Meeting
  • Place of Presentation: 米国 クリーブランド
  • Year and Date: 2014-10-12 – 2014-10-15
• [Presentation] Decreased Sox9 mRNA expression by cytokine stimulation was ameliorated by hyaluronan in chondrosarcoma cells (2014)
  • Author(s): Takashi Ohtsuki, Satoshi Hirohata, Yuri Shoji, Keiichi Asano, Aya Hirata, Teruyuki Kawadi, Junko Ingaki, Kanae Kumagishi, Keiichiro Nishida, Toshitaka Oohashi, Aiji Ohtsuka
  • Organizer: American Society for Matrix Biology Biennial Meeting
  • Place of Presentation: 米国 クリーブランド
  • Year and Date: 2014-10-12 – 2014-10-15
• [Presentation] 変形性膝関節症の治療効果に関する生化学的解析 (2014)
  • Author(s): 熊岸加苗、大月孝志、廣畑 聡、河村顕治、大塚愛二、坂田賢一郎、坂田俊輔
  • Organizer: 第27回日本臨床整形外科学会学術集会
  • Place of Presentation: 仙台市
  • Year and Date: 2014-07-20 – 2014-07-21
• [Presentation] 胎生期ADAMTS1ノックアウトマウスの解析 (2014)
  • Author(s): 楠絵里子、稲垣純子、大月孝志、川地輝幸、平田彩、浅野恵一、マティアス ホフマン、二宮善文、廣畑 聡
  • Organizer: 第46回日本結合組織学会学術大会 第61回マトリックス研究会大会
  • Place of Presentation: 名古屋市
  • Year and Date: 2014-06-05 – 2014-06-07
• [Presentation] 担がんマウスモデルにおけるバーシカン分解の解析 (2014)
  • Author(s): 浅野恵一、稲垣純子、マティアス ホフマン、川地輝幸、平田彩、大月孝志、二宮善文、廣畑 聡
  • Organizer: 第46回日本結合組織学会学術大会 第61回マトリックス研究会大会
  • Place of Presentation: 名古屋市
  • Year and Date: 2014-06-05 – 2014-06-07
• [Presentation] ヒアルロン酸短期投与によるラット変形性関節症モデルの関節軟骨保護効果の検討 (2014)
  • Author(s): 大月孝志、廣畑聡 、障子友理、浅野恵一 、平田彩 、川地輝幸、稲垣純子、熊岸加苗、西田圭一郎、二宮善文
  • Organizer: 第46回日本結合組織学会学術大会 第61回マトリックス研究会大会
  • Place of Presentation: 名古屋市
  • Year and Date: 2014-06-05 – 2014-06-07
• [Presentation] ADAMTS1はVEGFCと結合してリンパ管新生を抑制する (2014)
  • Author(s): 稲垣純子、高橋克之、小川弘子、浅野恵一、Omer Faruk Hatipoglu、 Mehmet Zeynel Cilek、小比賀真就、大月孝志、 Matthias Hofmann、 草地省蔵、二宮善文、廣畑聡
  • Organizer: 第46回日本結合組織学会学術大会 第61回マトリックス研究会大会
  • Place of Presentation: 名古屋市
  • Year and Date: 2014-06-05 – 2014-06-07
• [Presentation] 新規X線造影剤による変形性膝関節症ラット関節軟骨の高解像度マイクロＣＴ造影 (2014)
  • Author(s): 大橋俊孝、加来田博貴、芳谷 学、大月孝志、大野充昭、山田翔也、前原亜美、廣畑 聡、西田圭一郎、窪木拓男、二宮善文
  • Organizer: 第46回日本結合組織学会学術大会 第61回マトリックス研究会大会
  • Place of Presentation: 名古屋市
  • Year and Date: 2014-06-05 – 2014-06-07
• [Presentation] Cyclic tensile strain (CTS) down-regulates interleukin(IL)-1β and Tumor necrosis factor(TNF)-α induced matrix degrading enzymes mRNA expression in human chondrosarcoma cell line OUMS-27 by mechano sensor. (2014)
  • Author(s): Ohtsuki T, Kawadi T, Hirata A, Asano K, Inagaki J, Kumagishi K, Nishida K, Ninomiya Y, Hirohata S
  • Organizer: International symposium on mechanobiology 2014
  • Place of Presentation: 岡山市
  • Year and Date: 2014-05-20 – 2014-05-23
• [Presentation] Creation of Novel Articular Cartilage Specific X-ray Imaging Probes Composed of Lysine Oligomers. (2014)
  • Author(s): Oohashi T, Hagaya M, Ono M, Yamada S, Ohtsuki T, Maehara A, Hirohata S, Nishida K, Kuboki T, Kakuta H, NinomiyaY.
  • Organizer: International symposium on mechanobiology 2014
  • Place of Presentation: 岡山市
  • Year and Date: 2014-05-20 – 2014-05-23
• [Presentation] ADAMTS1 inhibits lymphangiogenesis by attenuating phosphorylation of the lymphatic endothelial cell-specific VEGF receptor (2014)
  • Author(s): Junko Inagaki, Katsuyuki Takahashi, Hiroko Ogawa, Omer Hatipoglu, Mehmet Cilek, Masanari Obika, Takashi Ohtsuki, Shozo Kusachi, Yoshifumi Ninomiya, Satoshi Hirohata
  • Organizer: The 18th International Vascular Biology Meeting
  • Place of Presentation: 京都市
  • Year and Date: 2014-04-14 – 2014-04-17