Investigation using 4-dimentional live imaging into the effects of aqueous cytokines on conjunctival scarring

• Project/Area Number: 15K15636
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Ophthalmology
• Research Institution: Kumamoto University
• Principal Investigator: Tanihara Hidenobu 熊本大学, 大学院生命科学研究部(医), 教授 (60217148)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 4次元イメージング / MCP-1 / 緑内障 / 生体分子 / 創傷治癒 / 生理活性

Outline of Final Research Achievements

The conditioned medium of conjunctival fibroblast cells increased migration of monocyte THP-1 cells. On the other hand, the conditioned medium of myofibroblast cells presented lower chemotactic activity of THP-1 migration in a comparison with medium of fibroblast. These migrations were inhibited by CCR2 inhibitor and MCP-1 neutralizing antibody. Additionally, the concentration of MCP-1 was decreased in conditioned medium of myofibroblast compared with conditioned medium of fibroblast.
We performed 4-dimentional live imaging using LysM-eGFP mice, and found that the number of LysM positive cell and the cell velocities were increased by MCP-1 administration to conjunctival surface. The effects of MCP-1 on conjunctiva, such as increased cell number and cell velocities, were suppressed by CCR2 inhibitor. These results indicated that LysM positive cells were accumulated and activated by MCP-1 through CCR2 receptor at conjunctiva.

Research Products

• [Journal Article] Molecular mechanisms underlying the filtration bleb-maintaining effects of suberoylanilide hydroxamic acid (SAHA). (2017)
  • Author(s): Futakuchi A, Inoue T, Fujimoto T, Kuroda U, Inoue-Mochita M, Takahashi E, Ohira S, Tanihara H.
  • Journal Title: Investigative Ophthalmology & Visual Sciences Volume: 58 Issue: 4 Pages: 2421-2429
  • DOI: 10.1167/iovs.16-21403
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Prospective observation of filtration blebs in high-risk eyes with limbal-based conjunctival flap. (2016)
  • Author(s): Matsumoto M, Inoue T, Kojima S, Nakashima KI, Fukushima A, Tanihara H.
  • Journal Title: Canadian Journal of Ophthalmology Volume: 51 Issue: 6 Pages: 431-437
  • DOI: 10.1016/j.jcjo.2016.05.019
  • Peer Reviewed
• [Journal Article] Change in visual acuity and associated risk factors after trabeculectomy with adjunctive mitomycin C. (2016)
  • Author(s): Kashiwagi K, Kogure S, Mabuchi F, Chiba T, Yamamoto T, Kuwayama Y, Araie M; Collaborative Bleb-Related Infection Incidence and Treatment Study Group.
  • Journal Title: Acta Ophthalmologica Volume: 94 Issue: 7 Pages: 561-570
  • DOI: 10.1111/aos.13058
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Comparing trabeculectomy outcomes between first and second operated eyes: A multicenter study. (2016)
  • Author(s): Iwasaki K, Takamura Y, Nishida T, Sawada A, Iwao K, Shinmura A, Kunimatsu-Sanuki S, Yamamoto T, Tanihara H, Sugiyama K, Nakazawa T, Inatani M.
  • Journal Title: PLOS ONE Volume: 11 Issue: 9 Pages: e0162569-e0162569
  • DOI: 10.1371/journal.pone.0162569
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The effects of ripasudil (K-115), a Rho kinase inhibitor, on activation of human conjunctival fibroblasts. (2016)
  • Author(s): Futakuchi A, Inoue T, Fujimoto T, Inoue-Mochita M, Kawai M, Tanihara H.
  • Journal Title: Experimental Eye Research Volume: 149 Pages: 107-115
  • DOI: 10.1016/j.exer.2016.07.001
  • Peer Reviewed / Open Access
• [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 Issue: 3 Pages: e0151947-e0151947
  • DOI: 10.1371/journal.pone.0151947
  • 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 Issue: 3 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): Tomokazu Fujimoto, Toshihiro Inoue, Miyuki Inoue-Mochita, Hidenobu Tanihara
  • Journal Title: Experimental Eye Research Volume: 145 Pages: 393-400
  • DOI: 10.1016/j.exer.2016.02.007
  • Peer Reviewed / Open Access
• [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 Issue: 1 Pages: 19640-19640
  • 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 Issue: 1 Pages: e0147080-e0147080
  • DOI: 10.1371/journal.pone.0147080
  • 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 Issue: 3 Pages: 288-290
  • 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 Issue: 1 Pages: 26-34
  • DOI: 10.1111/aos.12829
  • Peer Reviewed / Open Access
• [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 Issue: 10 Pages: e0139751-e0139751
  • DOI: 10.1371/journal.pone.0139751
  • Peer Reviewed / Open Access
• [Journal Article] In vivo imaging of mitochondrial axonal transport in the diseased and aged mammalian CNS (2015)
  • Author(s): Takihara et al.
  • Journal Title: Proceedings of the National Academy of Sciences of the U.S.A. Volume: 112 Issue: 33 Pages: 10515-10520
  • DOI: 10.1073/pnas.1509879112
  • Peer Reviewed / Open Access / Int'l Joint Research
• [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 Issue: 9 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 Issue: 6 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: 印刷中 Issue: 7 Pages: 755-761
  • DOI: 10.1001/jamaophthalmol.2015.0525
  • Peer Reviewed / Open Access
• [Presentation] Effects of interleukin (IL)-6 signaling activity on fibrogenic activity of human trabecular meshwork cells. (2016)
  • Author(s): Inoue T, Inoue-Mochita, M, Fujimoto T, Tanihara H.
  • Organizer: 12th European Glaucoma Society (EGS) Congress
  • Place of Presentation: Prague, Czech Republic
  • Year and Date: 2016-06-19
  • Int'l Joint Research
• [Presentation] Effects of interleukin (IL)-6 signaling activity on fibrogenic activity of human trabecular meshwork cells. (2016)
  • Author(s): Inoue T, Inoue-Mochita, M, Fujimoto T, Tanihara H.
  • Organizer: The Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO)
  • Place of Presentation: Seatle, Washington, USA
  • Year and Date: 2016-05-01
  • Int'l Joint Research
• [Presentation] Four-dimensional imaging revealed influences of monocyte chemoattractant protein-1 on immune cell dynamics in the subconjunctival tissue. (2016)
  • Author(s): Kojima S, Inoue T, Fujioto T, Tanihara H.
  • Organizer: The Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO)
  • Place of Presentation: Seatle, Washington, USA
  • Year and Date: 2016-05-01
  • Int'l Joint Research
• [Presentation] Effects of ripasudil (K-115), a Rho kinase inhibitor, on the activation of human conjunctival fibroblasts. (2016)
  • Author(s): Futakuchi A, Inoue T, Fujimoto T, Inoue-Mochita M, Tanihara H.
  • Organizer: The Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO)
  • Place of Presentation: Seatle, Washington, USA
  • Year and Date: 2016-05-01
  • Int'l Joint Research
• [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
• [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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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