4次元イメージングを用いた結膜瘢痕化におけるサイトカインの作用に関する研究

2015 Fiscal Year Research-status Report

• Project/Area Number: 15K15636
• Research Institution: Kumamoto University
• Principal Investigator: 谷原 秀信 熊本大学, 大学院生命科学研究部, 教授 (60217148)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: 生理活性 / 生体分子 / 緑内障

Outline of Annual Research Achievements

我々がこれまで行ってきた緑内障眼における房水中の生理活性物質のプロテオミクス的解析により、正常眼(白内障眼)や原発解放隅角緑内障眼と比較して、一般的に濾過手術成績が不良とされるぶどう膜炎続発緑内障・血管新生緑内障では顕著に房水中IL-6、IL-8、MCP-1濃度が高いことが示され、これらの炎症性サイトカインが緑内障濾過手術に負の影響を持つ可能性が示唆されている。
この結果を受けて、当該年度ではまず、MCP-1生理活性が結膜局所での炎症反応に与える効果について検証を行った。線維芽細胞由来と筋線維芽細胞由来の細胞培養上清が単核球細胞の遊走能に与える影響をTHP-1細胞株を用いて検証したところ、筋線維芽細胞由来上清の方が遊走能促進効果が弱く、筋線維芽細胞への分化により単核球を遊走が抑えられていることが明らかになった。またこれらTHP-1細胞の遊走は抗MCP-1中和抗体でベースラインまで抑制されることが分かり、MCP-1が本反応の主体をなしていることが明らかになった。次に、末梢組織でマクロファージが主に標識されるLysosome M(LysM)-eGFPマウスにアイカップを装着し、眼球結膜表面にMCP-1負荷を行った。その結果、定常状態と比較しMCP-1投薬後30-60分後にはLysM陽性細胞数が増加し、またその活動性を示す細胞速度が有意に増加していた。これらの結果から、MCP-1は眼内環境不均衡の代表的なマーカーであり、それ単独で結膜創傷治癒過程に強い影響を及ぼしうる可能性を示唆しているものと推測した。

Current Status of Research Progress

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

Reason

2光子顕微鏡を用いた動物術後モデルの結膜組織の4次元ライブイメージング法を確立するとともに、結膜炎症病態の解析を進行させている。
これまでに我々が行ってきた緑内障眼臨床サンプルを使用したプロテオミクス解析、臨床手術成績評価、in vitroおよびin vivoでの基礎研究のデータから得られた過剰な結膜創傷治癒過程を惹起しうる生理活性物質群の中から、主なターゲット分子としてMCP-1の存在を明らかとすることができた。以上の点から、順調に本研究が進捗しているものと判断する。

Strategy for Future Research Activity

本プロジェクトにより、緑内障眼においてMCP-1が眼内環境不均衡の代表的なマーカーであり、それ単独で結膜創傷治癒過程に強い影響を及ぼしうることが証明された。今後、MCP-1が緑内障治療における有力なターゲットとなりえるかについて検証をさらに進行させる。
眼内環境の病的状態からのリバランスを目指し、CCR2に着眼した研究を遂行する。CCR2はMCP-1を含めた多くのCCケモカインのレセプターであるため、CCR2阻害によるマクロファージ遊走能や活動性の抑制、さらには緑内障手術成績の改善の可能性について、in vivo緑内障手術モデルを用いた検証を予定する。

Research Products

• [Journal Article] The Influence of Phacoemulsification on Surgical Outcomes of Trabeculectomy with Mitomycin-C for Uveitic Glaucoma. (2016)
  • Author(s): Nishizawa A, Inoue T, Ohira S, Takahashi E, Saruwatari J, Iwao K, Tanihara H.
  • Journal Title: PLoS One Volume: 11 Pages: e0151947
  • DOI: 10.1371/journal.pone.0151947. eCollection 2016.
  • Peer Reviewed / Open Access
• [Journal Article] Visualization of Intravital Immune Cell Dynamics After Conjunctival Surgery Using Multiphoton Microscopy. (2016)
  • Author(s): Kojima S, Inoue T, Kikuta J, Furuya M, Koga A, Fujimoto T, Ueta M, Kinoshita S, Ishii M, Tanihara H.
  • Journal Title: Investigative Ophthalmology & Visual Science Volume: 57 Pages: 1207-1212
  • DOI: 10.1167/iovs.15-18507.
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Live cell imaging of actin dynamics in dexamethasone-treated porcine trabecular meshwork cells. (2016)
  • Author(s): Fujimoto T, Inoue T, Inoue-Mochita M, Tanihara H.
  • Journal Title: Experimental Eye Research Volume: 145 Pages: 393-400
  • DOI: 10.1016/j.exer.2016.02.007.
  • Peer Reviewed
• [Journal Article] Effects of K-115 (Ripasudil), a novel ROCK inhibitor, on trabecular meshwork and Schlemm's canal endothelial cells. (2016)
  • Author(s): Kaneko Y, Ohta M, Inoue T, Mizuno K, Isobe T, Tanabe S, Tanihara H.
  • Journal Title: Scientific Reports Volume: 6 Pages: 19640
  • DOI: 10.1038/srep19640.
  • Peer Reviewed / Open Access
• [Journal Article] Factors Influencing Aqueous Proinflammatory Cytokines and Growth Factors in Uveitic Glaucoma. (2016)
  • Author(s): Ohira S, Inoue T, Iwao K, Takahashi E, Tanihara H.
  • Journal Title: PLoS One Volume: 11 Pages: e0147080
  • DOI: 10.1371/journal.pone.0147080. eCollection 2016.
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Bleb-like Finding Caused by Idiopathic Scleral Thinning in an Eye With Absolute Glaucoma. (2016)
  • Author(s): Matsumura R, Inoue T, Tanihara H.
  • Journal Title: Journal of Glaucoma Volume: 25 Pages: e288-290
  • DOI: 10.1097/IJG.0000000000000341.
  • Peer Reviewed
• [Journal Article] One-year clinical evaluation of 0.4% ripasudil (K-115) in patients with open-angle glaucoma and ocular hypertension. (2016)
  • Author(s): Tanihara H, Inoue T, Yamamoto T, Kuwayama Y, Abe H, Fukushima A, Suganami H, Araie M; K-115 Clinical Study Group.
  • Journal Title: Acta Ophthalmologica Volume: 94 Pages: e26-34
  • DOI: 10.1111/aos.12829.
  • Peer Reviewed
• [Journal Article] Postoperative Changes in Aqueous Monocyte Chemotactic Protein-1 Levels and Bleb Morphology after Trabeculectomy vs. Ex-PRESS Shunt Surgery. (2015)
  • Author(s): Shobayashi K, Inoue T, Kawai M, Iwao K, Ohira S, Kojima S, Kuroda U, Nakashima K, Tanihara H.
  • Journal Title: PLoS One Volume: 10 Pages: e0139751
  • DOI: 10.1371/journal.pone.0139751. eCollection 2015.
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] In vivo imaging of axonal transport of mitochondria in the diseased and aged mammalian CNS. (2015)
  • Author(s): Takihara Y, Inatani M, Eto K, Inoue T, Kreymerman A, Miyake S, Ueno S, Nagaya M, Nakanishi A, Iwao K, Takamura Y, Sakamoto H, Satoh K, Kondo M, Sakamoto T, Goldberg JL, Nabekura J, Tanihara H.
  • Journal Title: Proceedings of the National Academy of Sciences of the United States of America Volume: 112 Pages: 10515-10520
  • DOI: 10.1073/pnas.1509879112.
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Effect of laser suture lysis on filtration openings: a prospective three-dimensional anterior segment optical coherence tomography study. (2015)
  • Author(s): Cho HK, Kojima S, Inoue T, Fukushima A, Kee C, Tanihara H.
  • Journal Title: Eye Volume: 29 Pages: 1220-1225
  • DOI: 10.1038/eye.2015.129.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Simultaneous increase in multiple proinflammatory cytokines in the aqueous humor in neovascular glaucoma with and without intravitreal bevacizumab injection. (2015)
  • Author(s): Ohira S, Inoue T, Shobayashi K, Iwao K, Fukushima M, Tanihara H.
  • Journal Title: Investigative Ophthalmology & Visual Science Volume: 56 Pages: 3541-3548
  • DOI: 10.1167/iovs.14-15918.
  • Peer Reviewed / Open Access
• [Journal Article] Additive Intraocular Pressure-Lowering Effects of the Rho Kinase Inhibitor Ripasudil (K-115) Combined With Timolol or Latanoprost: A Report of 2 Randomized Clinical Trials. (2015)
  • Author(s): Tanihara H, Inoue T, Yamamoto T, Kuwayama Y, Abe H, Suganami H, Araie M; K-115 Clinical Study Group.
  • Journal Title: JAMA Ophthalmology Volume: 133 Pages: 755-761
  • DOI: 10.1001/jamaophthalmol.2015.0525.
  • Peer Reviewed
• [Presentation] The effect of phacoemulsification on failure of trabeculectomy in uveitic glaucoma patients. (2015)
  • Author(s): Nishizawa A, Takahashi E, Ohira S, Iwao K, Inoue T, Tanihara H.
  • Organizer: XXXIII Congress of the European Society of Cataract & Refractive Surgeons (ESCRS)
  • Place of Presentation: Barcelona, Spain
  • Year and Date: 2015-09-05 – 2015-09-09
• [Presentation] Two cases with scleral fixation of intraocular lens after trabeculectomy. (2015)
  • Author(s): Takahashi E, Nishizawa A, Inoue T, Kawabata K. Tanihara H.
  • Organizer: XXXIII Congress of the European Society of Cataract & Refractive Surgeons (ESCRS)
  • Place of Presentation: Barcelona, Spain
  • Year and Date: 2015-09-05 – 2015-09-09
  • Int'l Joint Research
• [Presentation] Impact of phacoemulsification on postoperative IOP after MMC trabeculectomy for uveitic glaucoma. (2015)
  • Author(s): Iwao K, Nishizawa A, Inoue T, Tanihara H.
  • Organizer: XXXIII Congress of the European Society of Cataract & Refractive Surgeons (ESCRS)
  • Place of Presentation: Barcelona, Spain
  • Year and Date: 2015-09-05 – 2015-09-09
  • Int'l Joint Research
• [Presentation] Imaging for Glaucoma Diagnosis and Monitoring: New Idea from Asia, Impact of anterior segment OCT on bleb evaluation. (2015)
  • Author(s): Inoue T.
  • Organizer: The 6th World Glaucoma Congress
  • Place of Presentation: Hong Kong, China
  • Year and Date: 2015-06-06 – 2015-06-09
  • Int'l Joint Research
• [Presentation] Rho kinase inhibitors as a new glaucoma medication, Presentation title: The role of actomyosin system in regulating aqueous outflow. (2015)
  • Author(s): Inoue T.
  • Organizer: The 6th World Glaucoma Congress
  • Place of Presentation: Hong Kong, China
  • Year and Date: 2015-06-06 – 2015-06-09
  • Int'l Joint Research
• [Presentation] Histone deacetylase inhibitor attenuates TGF-β2 induced human conjunctival fibroblast activation. (2015)
  • Author(s): Futakuchi A, Inoue T, Fujimoto T, Kuroda U, Inoue-Mochita M, Takahashi E, Shobayashi K, Ohira S, Kojima S, Tanihara H.
  • Organizer: The Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO)
  • Place of Presentation: Denver, Colorado, USA
  • Year and Date: 2015-05-03 – 2015-05-07
  • Int'l Joint Research
• [Presentation] The effect of VEGF-A on trabecular meshwork and Schlemm’s canal endothelial cells permeability. (2015)
  • Author(s): Fujimoto T, Inoue T, Kei Maki, Inoue-Mochita M, Kojima S, Tanihara H.
  • Organizer: The Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO)
  • Place of Presentation: Denver, Colorado, USA
  • Year and Date: 2015-05-03 – 2015-05-07
  • Int'l Joint Research
• [Presentation] TGF--activated kinase 1 is essential for tumor necrosis factor--induced epithelial mesenchymal transition in ARPE-19 cells. (2015)
  • Author(s): Matsubara K, Takahashi E, Haga A and Tanihara H.
  • Organizer: The Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO)
  • Place of Presentation: Denver, Colorado, USA
  • Year and Date: 2015-05-03 – 2015-05-07
  • Int'l Joint Research
• [Presentation] Merlin regulates hyaluronan endocytosis and epithelial mesenchymal transition in ARPE-19 cells. (2015)
  • Author(s): Takahashi E, Haga A and Tanihara H.
  • Organizer: The Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO)
  • Place of Presentation: Denver, Colorado, USA
  • Year and Date: 2015-05-03 – 2015-05-07
  • Int'l Joint Research
• [Presentation] The subtypes of microglia in rat after optic nerve crush injury. (2015)
  • Author(s): Haga A, Takahashi E, Inomata Y, Tanihara H.
  • Organizer: The Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO)
  • Place of Presentation: Denver, Colorado, USA
  • Year and Date: 2015-05-03 – 2015-05-07
  • Int'l Joint Research
• [Remarks] 熊本大学大学院生命科学研究部眼科学分野 研究内容
  • URL: http://www2.kuh.kumamoto-u.ac.jp/ganka/kyousitu/naiyou.html