線維化におけるＴＧＦ－β情報伝達の関与についての分子基盤の解明

2012 Fiscal Year Research-status Report

• Project/Area Number: 24591658
• Research Category: Grant-in-Aid for Scientific Research (C)
• Research Institution: Kumamoto University
• Principal Investigator: 尹 浩信 熊本大学, その他の研究科, 教授 (20282634)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: 遺伝子 / 細胞・組織 / シグナル伝達

Research Abstract

全身性強皮症、肺線維症の組織および病変部の培養線維芽細胞においてDNA microarry法にて過剰発現する分子、発現低下する分子を検討し、またrealtime PCRを用いたmicro RNA arrayによって過剰発現するmicro RNA、発現低下するmicro RNAを同定した。
正常皮膚線維芽細胞を用いてintegrin αVβ5 stable transfectant作成し、integrin αVβ5の過剰発現を確認した。
COL1A2 enhancer、promoterをベクターに組み込んで作成したtransgeneを用いて線維芽細胞特異的integrin αVβ5 transgenic mouseを作成すし、現在phenotypeを解析している。

Current Status of Research Progress

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

Reason

線維化病変に過剰発現ないし発現低下する分子を同定することができた。さらに過剰発現するmicro RNA、発現低下するmicro RNAをも同定することができた。integrin αVβ5 stable transfectantも作成でき、線維芽細胞特異的integrin αVβ5 transgenic mouseも完成した。
以上より研究は順調で十分達成できたと考える。

Strategy for Future Research Activity

integrin αVβ5 stable transfectantを用いてDNA microarry法、realtime PCRを用いたmicro RNA array法、情報伝達protein arrayを行い、線維化phenotypeを形成しているか、すなわちTGF-β受容体が過剰発現しているか、TGF-β情報伝達経路（Smad2、Smad 3、Smad4、p300/CBP、p38 MAPK、PI3 kinase）が過剰リン酸化しているか、I型コラーゲンをはじめとする細胞外マトリックス蛋白および遺伝子が過剰発現しているか、MMP-1などのproteaseの発現が低下しているか、TIMP-1などのprotease inhibitorの発現が亢進しているか詳細に検討する。
線維芽細胞特異的integrin αV transgenic mouseのphenotypeを解析する。

Expenditure Plans for the Next FY Research Funding

integrin αVβ5 stable transfectantの解析および線維芽細胞特異的integrin αV transgenic mouseのphenotypeの解析に用いる研究試薬に全額使用する。

Research Products

• [Journal Article] miR-150 down-regulation contributes to the constitutive type I collagen overexpression in scleroderma dermal fibroblasts via the induction of integrin β3. (2013)
  • Author(s): Honda N, Jinnin M, Kira-Etoh T, Makino K, Kajihara I, Makino T, Fukushima S, Inoue Y, Okamoto Y, Hasegawa M, Fujimoto M, Ihn H.
  • Journal Title: Am J Pathol, Volume: 182 Pages: 206-216
  • Peer Reviewed
• [Journal Article] The rs2910164 G>C polymorphism in microRNA-146a is associated with the incidence of malignant melanoma. (2013)
  • Author(s): Yamashita J, Iwakiri T, Fukushima S, Jinnin M, Miyashita A, Hamasaki T, Makino T, Aoi J, Masuguchi S, Inoue Y, Ihn H.
  • Journal Title: Melanoma Res, Volume: 23 Pages: 13-20
  • Peer Reviewed
• [Journal Article] The miR-146a rs2910164G/C polymorphism is associated with telangiectasia in systemic sclerosis. (2013)
  • Author(s): Sakoguchi A, Jinnin M, Makino T, Kajihara I, Makino K, Honda N, Nakayama W, Inoue K, Fukushima S, Ihn H.
  • Journal Title: Clin Exp Dermatol, Volume: 38 Pages: 99-100
  • Peer Reviewed
• [Journal Article] microRNA-7 down-regulation mediates excess collagen expression in localized scleroderma. (2013)
  • Author(s): Etoh M, Jinnin M, Makino K, Yamane K, Nakayama W, Aoi J, Honda N, Kajihara I, Makino T, Fukushima S, Ihn H.
  • Journal Title: Arch Dermatol Res, Volume: 305 Pages: 9-15
  • Peer Reviewed
• [Journal Article] Down-regulation of miR-124/ -214 in cutaneous squamous cell carcinoma mediates abnormal cell proliferation via the induction of ERK. (2013)
  • Author(s): Yamane K, Jinnin M, Etoh T, Kobayashi Y, Shimozono N, Fukushima S, Masuguchi S, Maruo K, Inoue Y, Ishihara T, Aoi J, Oike Y, Ihn H.
  • Journal Title: J Mol Med, Volume: 91 Pages: 69-81
  • Peer Reviewed
• [Journal Article] Intravenous immunoglobulin treatment recovers the down-regulatied levels of Th1 cytokines in the sera and skin of scleroderma patients. (2013)
  • Author(s): Kudo H, Jinnin M, Yamane K, Makino T, Kajihara I, Makino K, Honda N, Nakayama W, Fukushima S, Ihn H.
  • Journal Title: J Dermatol Sci, Volume: 69 Pages: 77-80
  • Peer Reviewed
• [Journal Article] Serum epidermal growth factor receptor levels in patients with malignant melanoma. (2013)
  • Author(s): Kanemura H, Fukushima S, Yamashita J, Jinnin M, Sakai K, Masuguchi M, Aoi J, Makino T, Inoue Y, Ihn H.
  • Journal Title: Clin Exp Dermatol, Volume: 38 Pages: 172-177
  • Peer Reviewed
• [Journal Article] Serum miR-21 levels in patients with dermatomyositis. (2013)
  • Author(s): Shimada S, Jinnin M, Ogata A, Makino T, Kajihara I, Makino K, Honda N, Nakayama W, Inoue K, Fukushima S, Ihn H.
  • Journal Title: Clin Exp Rheumatol, Volume: 31 Pages: 161-162
  • Peer Reviewed
• [Journal Article] Serum levels of interleukin-1α in patients with systemic sclerosis. (2013)
  • Author(s): Maekawa T, Jinnin M, Ohtsuki M, Ihn H.
  • Journal Title: J Dermatol, Volume: 40 Pages: 98-101
  • Peer Reviewed
• [Journal Article] Suppresion of UV-induced wrinkle formation by induction of HSP70 expression in mice. (2013)
  • Author(s): Matsuda M, Hoshino T, Yamakawa N, Tahara K, Adachi H, Sobue G, Maji D, Ihn H, Mizushima T.
  • Journal Title: J Invest Dermatol, Volume: 133 Pages: 919-928
  • Peer Reviewed
• [Journal Article] Pyodermatitis vegetans associated with multiple myeloma. (2013)
  • Author(s): Kajihara I, Ichihara A, Higo J, Kidou M, Ihn H.
  • Journal Title: J Dermatol, Volume: 40 Pages: 222-223
  • Peer Reviewed
• [Journal Article] Diccoidin domain receptor 2 -micro RNA 196a -mediated negative feedback against excess type I collagen expression is impaired in scleroderma dermal fibroblasts. (2013)
  • Author(s): Makino K, Jinnin M, Aoi J, Hirano A, Kajihara I, Makino T, Sakai K, Fukushima S, Inoue Y, Ihn H.
  • Journal Title: J Invest Dermatol, Volume: 133 Pages: 110-119
  • Peer Reviewed
• [Journal Article] Decreased miR-7 expression in the skin and sera of patients with dermatomyositis. (2013)
  • Author(s): Oshikawa Y, JInnin M, Makino T, Kajihara I, Makino K,Honda N, Nakayama W, Inoue K, Fukushima S, Ihn H.
  • Journal Title: Acta Derm Venereol Volume: 4 Pages: 144‐149
  • Peer Reviewed
• [Journal Article] Hidradenocarcinoma showing prominent mucinous and squamous differentiation and associated with pgetoid cells. (2013)
  • Author(s): Honda Y, Tanigawa H, Harada M, Fukushima S, Masuguchi S, Ishihara T, Ihn H, Iyama KI.
  • Journal Title: J Cutan Pathol, Volume: 印刷中 Pages: 印刷中
  • Peer Reviewed
• [Journal Article] Dots/ globules on dermoscopy in nail-apparatus melanoma. (2013)
  • Author(s): Inoue Y, Menzies SW, Fukushima S, Nishi-Kogushi H, Miyashita A, Masuguchi S, Muchemwa F, Kageshita T, Ihn H.
  • Journal Title: Int J Dermatol, Volume: 印刷中 Pages: 印刷中
  • Peer Reviewed
• [Journal Article] Scleroderma dermal fibroblasts overexpress vascular endothelial growth factor due to autocrine transforming growth factor  signaling. (2013)
  • Author(s): Kajihara I, Jinnin M, Honda N, Makino K, Makino T, Masuguchi S, Sakai K, Fukushima S, Inoue Y, Ihn H.
  • Journal Title: Mod Rheumatol, Volume: 印刷中 Pages: 印刷中
  • Peer Reviewed
• [Journal Article] Serum chemokine levels as prognostic markers in patients with early systemic sclerosis: a multicentre, prospective, observational study. (2013)
  • Author(s): Hasegawa M, Asano M, Endo H, Fujimoto M, Goto D, Ihn H, Inoue K, Ishikawa O, Kawaguchi Y, Kuwana M, Ogawa F, Takahashi H, Tanaka S, Sato S, Takehara K.
  • Journal Title: Mod Rheumatol, Volume: 印刷中 Pages: 印刷中
  • Peer Reviewed
• [Journal Article] Case of paraneoplastic pemphigus with follicular lymphoma treated with rituximab. (2013)
  • Author(s): Aoi J, Makino K, Sakai k, Masuguchi S, Fukushima S, Jinnin M, Inoue Y, Koga H, Hashimoto T, Ihn H.
  • Journal Title: J Dermatol, Volume: 40 Pages: 285-286
  • Peer Reviewed
• [Journal Article] The down-regulation of microRNA let-7a contributes to excessive expression of type I collagen in systemic and localized scleroderma. (2013)
  • Author(s): Makino K, Jinnin M, Hirano A, Yamane K, Eto M, Kusano T, Honda N, Kajihara K, Makino T, Sakai K, Masuguchi S, Fukushima S, Ihn H.
  • Journal Title: J Immunol, Volume: 190 Pages: 3905-3915
  • Peer Reviewed
• [Journal Article] Clinical significance of cobble stone appearance on the skin of patients with systemic sclerosis. (2012)
  • Author(s): Kajihara I, Jinnin M, Makino T, Toihata-Yonemitsu A, Sakai K, Masuguchi S, Fukushima S, Ihn H.
  • Journal Title: J Eur Acad Dermatol Venereol, Volume: 26 Pages: 123－125
  • Peer Reviewed
• [Journal Article] Investigation of prognostic factors for skin sclerosis and lung function in Japanese patients with systemic sclerosis: a multicenter prospective observational study. (2012)
  • Author(s): Hasegawa M, Asano Y, Endo H, Fujimoto M, Goto D, Ihn H, Inoue K, Ishikawa O, Kawaguchi Y, Kuwana M, Muro Y, Ogawa F, Sasaki T, Takahashi H, Tanaka S, Takehara K, Sato S.
  • Journal Title: Rheumatology, Volume: 51 Pages: 129－133
  • Peer Reviewed
• [Journal Article] Prediction of additional lymph node positivity and clinical outcome of micrometastases in sentinel lymph nodes in cutaneous melanoma: A multi-institutional study of 450 patients in Japan. (2012)
  • Author(s): Namikawa K, Yamazaki N, Nakai Y, Ihn H, Tomita Y, Uhara H, Takenouchi T, Kiyohara Y, Moroi Y, Yamamoto Y, Otsuka F, Kamiya H, Iizuka H, Hatta N, Kadono T.
  • Journal Title: J Dermatol, Volume: 39 Pages: 130－137
  • Peer Reviewed
• [Journal Article] Serum levels of soluble CD21 in patients with sclerosis. (2012)
  • Author(s): Tomita M, Kadono T, Yazawa N, Kawashima T, Tamaki Z, Ashida R, Ohmatsu H, Asano Y, Sugaya M, Kubo M, Ihn H, Tamaki K, Sato S.
  • Journal Title: Rheumatol Int, Volume: 32 Pages: 317－321
  • Peer Reviewed
• [Journal Article] Serum levels of soluble CD163 in patients with systemic sclerosis. (2012)
  • Author(s): Nakayama W, Jinnin M, Makino K, Kajihara I, Makino T, Fukushima S, Inoue Y, Ihn H.
  • Journal Title: Rheumatol Int, Volume: 32 Pages: 403－407
  • Peer Reviewed
• [Journal Article] Increased accumulation of thrombospondin-2 due to low degradation activity stimulates type I collagen expression in scleroderma fibroblasts. (2012)
  • Author(s): Kajihara I, Jinnin M, Yamane K, Makino T, Honda N, Igata T, Masuguchi S, Fukushima S, Okamoto Y, Hasegawa M, Fujimoto M, Ihn H.
  • Journal Title: Am J Pathol, Volume: 180 Pages: 703－714
  • Peer Reviewed
• [Journal Article] Cutaneous type adult T-cell leukemia/lymphoma successfully treated with narrow-band ultraviolet B phototherapy. (2012)
  • Author(s): Kudo K, Fukushima S, Masuguchi S, Sakai K, Jinnin M, Ihn H.
  • Journal Title: Clin Exp Dermatol, Volume: 37 Pages: 183－184
  • Peer Reviewed
• [Journal Article] The role of angiopoietin-like protein 2 in pathogenesis of dermatomyositis. (2012)
  • Author(s): Ogata A, Endo M, Aoi J, Takahashi O, Kadomatsu T, Miyata K, Tian Z, Jinnin M, Fukushima S, Ihn H, Oike Y.
  • Journal Title: Biochem Biophys Res Commun, Volume: 418 Pages: 494－499
  • Peer Reviewed
• [Journal Article] Circulating miR-142-3p levels in patients with systemic sclerosis. (2012)
  • Author(s): Makino K, Jinnin M, Kajihara I, Honda N, Sakai K, Masuguchi S, Fukushima S, Inoue Y, Ihn H.
  • Journal Title: Clin Exp Dermatol, Volume: 37 Pages: 34－39
  • Peer Reviewed
• [Journal Article] TGF-β-mediated down-regulation of microRNA-196a contributes to the constitutive up-regulated type I collagen expression in scleroderma dermal fibroblasts. (2012)
  • Author(s): Honda N, Jinnin M, Kajiwara I, Makino T, Makino K, Masuguchi S, Fukushima S, Okamoto Y, Hasegawa M, Fujimoto M, Ihn H.
  • Journal Title: J Immunol, Volume: 188 Pages: 3323－3331
  • Peer Reviewed
• [Journal Article] Increased serum levels of miR-1266 in patients with psoriasis vulgaris. (2012)
  • Author(s): Ichihara A, Jinnin M, Oyama R, Yamane K, Fujisawa A, Sakai K, Masuguchi S, Fukushima S, Maruo K, Ihn H.
  • Journal Title: Eur J Dermatol, Volume: 22 Pages: 68－71
  • Peer Reviewed
• [Journal Article] Impaired IL-17 signaling pathway contributes to the increased collagen expression in scleroderma fibroblasts. (2012)
  • Author(s): Nakajima T, Jinnin M, Honda N, Kajihara I, Makino T, Masuguchi S, Fukushima S, Okamoto Y, Hasegawa M, Fujimoto M, Ihn H.
  • Journal Title: J Immunol, Volume: 188 Pages: 3573-3583
  • Peer Reviewed
• [Journal Article] Photosensitivity and acute liver insufficiency in late-onset erythropoietic protoporphyria with a chromosome 18q abnormality. (2012)
  • Author(s): Oshikawa Y, Fukushima S, Miyake T, Kawaguchi T, Motomura K, Nakashima Y, Nakamura K, Jinnin M, Ihn H.
  • Journal Title: Case Rep Dermatol Volume: ４ Pages: 14-149
  • Peer Reviewed
• [Journal Article] Overexpression of hepatocyte growth factor receptor in scleroderma dermal fibroblasts is caused by autocrine transforming growth factor βsignaling. (2012)
  • Author(s): Kajihara I, Jinnin M, Makino T, Masuguchi S, Sakai K, Fukushima S, Maruo K, Inoue Y, Ihn H.
  • Journal Title: BioScience Trends, Volume: 6 Pages: 136－142
  • Peer Reviewed
• [Journal Article] microR-92a expression in the sera and dermal fibroblasts increases in patients with scleroderma. (2012)
  • Author(s): Sing T, Jinnin M, Yamane K, Honda N, Makino K, Kajihara I, Makino T, Sakai K, Mashuguchi S, Fukushima S, Ihn H.
  • Journal Title: Rheumatology, Volume: 51 Pages: 1550-1556
  • Peer Reviewed
• [Journal Article] Kinase family member 20A as is a novel melanoma-associated antigen. (2012)
  • Author(s): Yamashita J, Fukushima S, Jinnin M, Honda N, Makino K, Sakai K, Masuguchi S, Inoue Y, Ihn H.
  • Journal Title: Acta Derm Venereol, Volume: 92 Pages: 593-597
  • Peer Reviewed
• [Journal Article] CD163 expression is increased in the involved skin and sera of patients with systemic lupus erythematosus. (2012)
  • Author(s): Nakayama W, Jinnin M, Makino K, Kajihara I, Makino T, Fukushima S, Sakai k, Inoue Y, Ihn H.
  • Journal Title: Eur J Dermatol, Volume: 22 Pages: 512-517
  • Peer Reviewed
• [Journal Article] Serum angiopoietin-like protein 3 concentrations in rheumatic diserases. (2012)
  • Author(s): Hayashi Y, Jinnin M, Makino T, Kajihara I, Makino K, Honda N, Nakayama W, Inoue K, Fukushima S, Ihn H.
  • Journal Title: Eur J Dermatol, Volume: 22 Pages: 500-504
  • Peer Reviewed
• [Journal Article] Diagnostic and prognostic value of subcutaneous tissue biopsy in patients with cardiac amyloidosis. (2012)
  • Author(s): Takashio S, Izumiya Y, Jinnin M, Yamamuro M, Kojima S, Ihn H, Ogawa H.
  • Journal Title: Am J Cardiol, Volume: 110 Pages: 1507-1511
  • Peer Reviewed
• [Journal Article] Tsukushi controls the hair cycle by regulating TGF-β1 signaling. (2012)
  • Author(s): Niimori D, Kawano R, Felemban A, Niimori-Kita K, Tanaka H, Ihn H, Ohta K.
  • Journal Title: Dev Biol, Volume: 372 Pages: 81-87
  • Peer Reviewed
• [Journal Article] Case of blastic plasmaytoid dendritic cell neoplasm. (2012)
  • Author(s): Aoi J, Ogata A, Makino T, Sakai K, Masuguchi S, Fukushima S, Jinnin M, Inoue Y, Ihn H.
  • Journal Title: J Dermatol, Volume: 39 Pages: 1066-1067
  • Peer Reviewed
• [Journal Article] Serum concentration of angiogenin in patients with rheumatic diseases. (2012)
  • Author(s): Kuwahara A, Jinnin M, Makino T, Kajihara I, Makino K, Honda N, Nakayama W, Inoue K, Fukushima S, Ihn H.
  • Journal Title: BioScience Trends, Volume: 6 Pages: 229-233
  • Peer Reviewed
• [Presentation] Interleukin-23 signaling contributes to the increased collagen production in scleroderma fibroblasts. (2012)
  • Author(s): Nakayama W, Jinnin M, Fukushima S, Ihn H.
  • Organizer: The 37 th Annual Meeting of the Japanese Society for Investigative Dermatology
  • Place of Presentation: ロワジールホテル＆スパタワー那覇（Okinawa）
  • Year and Date: 20121207-20121209
• [Presentation] microRNA-150 down-regulation contributes to the constitutive type I collagen overexpression in scleroderma dermal fibroblasts via the induction of integrinβ3. (2012)
  • Author(s): Honda N, Jinnin M, Makino K, Makino T, Fukushima S, Inoue Y, Okamoto Y, Hasegawa M, Fujimoto M, Ihn H.
  • Organizer: The 37 th Annual Meeting of the Japanese Society for Investigative Dermatology
  • Place of Presentation: ロワジールホテル＆スパタワー那覇（Okinawa）
  • Year and Date: 20121207-20121209
• [Presentation] Excessive expression of type I collagen in systemic and localized scleroderma is caused by the down-regulation of microRNA let-7a. (2012)
  • Author(s): Makino K, Jinnin M, Yamane K, Eto M, Kusano T, Honda N, Kajihara I, Makino T, Sakai K, Masuguchi S, Fukushima S, Ihn H.
  • Organizer: The 37 th Annual Meeting of the Japanese Society for Investigative Dermatology
  • Place of Presentation: ロワジールホテル＆スパタワー那覇（Okinawa）
  • Year and Date: 20121207-20121209
• [Presentation] Intravenous immunoglobulin treatment recovers the down-regulated levels of Th1 cytokines in the sera and skin of scleroderma patients. (2012)
  • Author(s): Kudo H, Jinnin M, Yamane K, Makino T, Kajihara I, Makino K, Honda N, Nakayama W, Fukushima S, Ihn H.
  • Organizer: The 37 th Annual Meeting of the Japanese Society for Investigative Dermatology
  • Place of Presentation: ロワジールホテル＆スパタワー那覇（Okinawa）
  • Year and Date: 20121207-20121209
• [Presentation] Decreased adiponectin levels in the patients with diffuse cutaneous systemic sclerosis. (2012)
  • Author(s): Fukushima S, Jinnin M, Arakawa H, Makino T, Kajihara I, Makino K, Honda N, Inoue Y, Ihn H.
  • Organizer: The 37 th Annual Meeting of the Japanese Society for Investigative Dermatology
  • Place of Presentation: ロワジールホテル＆スパタワー那覇（Okinawa）
  • Year and Date: 20121207-20121209
• [Presentation] Dysregulated discoidin domain receptor 2-microRNA196a ‐mediated negative feedback against excess type I collagen expression in scleroderma dermal fibroblasts. (2012)
  • Author(s): Makino K, Jinnin M, Aoi J, Kajihara I, Makino T, Sakai K, Fukushima S, Inoue Y, Ihn H.
  • Organizer: American College of Rheumatology
  • Place of Presentation: Washington D.C.（アメリカ）
  • Year and Date: 20121109-20121114
• [Presentation] microRNA-150 down-regulation contributes to the constitutive type I collagen overexpression in scleroderma dermal fibroblasts via the induction of integrin β3. (2012)
  • Author(s): Honda N, JInnin M, Makino K, Makino T, Fukushima S, Inoue Y, Okamoto Y, Hasegawa M, Fujimoto M, Ihn H.
  • Organizer: 42 nd annual ESDR Meeting
  • Place of Presentation: Venice（イタリア）
  • Year and Date: 20120919-20120922
• [Presentation] The levels of adiponectin are decreased in the skin and sera of diffuse cutaneous systemic sclerosis patients. (2012)
  • Author(s): Kubo Y, Fukushima S, Jinnin M, Makino T, Kajihara I, Ichihara A, Makino K, Ihn H.
  • Organizer: Annual European Congress of Rheumatology
  • Place of Presentation: Berlin(ドイツ）
  • Year and Date: 20120606-20120609