Cornea - ARVO

ARVO 2013 Annual Meeting Abstracts by Scientific Section/Group - CorneaLu Chen, Sammy Grimaldo, Tatiana Ecoiffier, Don Yuen. Center forEye Disease & Development, Program in Vision Science and Schoolof Optometry, University of California, Berkeley, CA.Purpose: MicroRNAs are a class of small non-coding RNAs thatnegatively regulate gene expression by binding to complimentarysequences of target messenger RNAs. Their roles in the cornea stillremain largely unknown. Here we present our new findings on theanti-lymphangiogenesis function of microRNA-184 in the corneaduring an inflammatory response in vivo.Methods: The murine in vivo suture placement model was used toinduce corneal inflammatory lymphangiogenesis (LG). Mice wererandomly selected to receive subconjunctival injections of eithersynthetic microRNA-184 mimics or control twice a week after theprocedure. Whole-mount corneas were sampled and stained withLYVE-1, the lymphatic marker, for immunofluorescent microscopicassays. Results were analyzed by NIH-ImageJ software.Results: Compared to the control condition, the lymphatic invasionarea was significantly reduced after the treatment of MicroRNA-184mimics in the inflamed cornea.Conclusions: MicroRNA-184 suppresses corneallymphangiogenesis. Its further investigation may provide noveltherapies for lymphatic related disorders in the cornea, such asinflammation and transplant rejection.Commercial Relationships: Lu Chen, None; Sammy Grimaldo,None; Tatiana Ecoiffier, None; Don Yuen, NoneSupport: This work is supported in part by research grants from NIHand University of California at Berkeley (LC).Program Number: 2096 Poster Board Number: D0235Presentation Time: 11:00 AM - 12:45 PMLive Imaging of Lymphatic Valve Formation after CornealTransplantationGyeong Jin Kang 1 , Tatiana Ecoiffier 1 , Young-Kwon Hong 2 , LuChen 1 . 1 Center for Eye Disease and Development, Program in VisionScience and School of Optometry, University of California, Berkeley,CA; 2 Department of Surgery, Norris Comprehensive Cancer Center,University of Southern California, Los Angeles, CA.Purpose: The lymphatic pathway is a major mediator for transplantrejection. Most recently, we have provided the first evidence onlymphatic valve formation during corneal inflammatorylymphangiogenesis (Truong et al. 2011). The purpose of this realtimein vivo study is to investigate the time course and pattern oflymphangiogenesis as well as lymphatic valve formation induced bycorneal transplantation with our newly developed live imagingsystem and Prox-1-GFP mice.Methods: Standard orthotopic corneal transplantation was performedwith Prox1-GFP mice as recipients. Corneal grafts of the same micewere continuously observed by the live imaging system forlongitudinal analysis. Prox-1 positive lymphatic vessels and valveswere evaluated at both limbal and corneal areas.Results: Prox-1 positive lymphatic vessels and valves were formedafter corneal transplantation. As corneal lymphangiogenesisproceeded, more valves were observed inside vessel cavities.Furthermore, lymphatic valvulogenesis was initiated inside limbalvessels before spreading into the central cornea.Conclusions: We have shown, for the first time, cornealtransplantation induces both lymphangiogenesis and valvulogenesisin vivo and in real time. Further investigation on this newphenomenon may reveal new mechanisms underlying transplantrejection. Since the lymphatic system plays an important role in manyother functions, this study may also offer new insights to ourunderstanding and treatment of other lymphatic related disordersoutside the eye.Commercial Relationships: Gyeong Jin Kang, None; TatianaEcoiffier, None; Young-Kwon Hong, None; Lu Chen, NoneSupport: This work is supported in part by research grants from NIHand University of California at Berkeley (LC).Program Number: 2097 Poster Board Number: D0236Presentation Time: 11:00 AM - 12:45 PMNovel Characterization of MHC-Class II Positive Cells inEmbryonic Cornea during Spontaneous Lymphatic EventsDon Yuen, Guangyu Li, Lu Chen. Center for Eye Disease andDevelopment, Program in Vision Science and School of Optometry,University of California, Berkeley, CA.Purpose: We recently reported that unlike the adult cornea which isdevoid of lymphatic vessels, the immature cornea is supplied bylymphatics which undergo spontaneous regression duringdevelopment. Since lymphatic vessels mediate antigen presenting celltrafficking during an immune response, this study is to investigatewhether the distribution of dendritic cells in immature cornea differsfrom the adult stage and how it correlates with the lymphatic events.Methods: Corneal samples of different developmental stages wereisolated from C57B6 mice and stained with a panel of specificmarkers for MHC-Class II, CD45, CD11c, and LYVE-1 forimmunofluorescent microscopic assays.Results: In contrast to the adult cornea where MHC Class II positivecells were found in the peripheral area, embryonic cornea at E16.5also exhibited these cells in the central area. The cells expressedCD45 and CD11c as well. As the cornea became mature andunderwent spontaneous lymphatic formation and regression, theMHC Class II positive cells were more restricted to the peripheralarea.Conclusions: Our findings have shown, for the first time, MHCClass II positive cells are present in the central cornea duringdevelopment. Further investigation on this new phenomenon mayprovide novel insights into corneal embryogenesis as well as thecreation and regulation of immune privilege in the cornea.Commercial Relationships: Don Yuen, None; Guangyu Li, None;Lu Chen, NoneSupport: This work is supported in part by research grants from NIHand University of California at Berkeley (LC)Program Number: 2098 Poster Board Number: D0237Presentation Time: 11:00 AM - 12:45 PMDeletion of Foxc1 and/or Foxc2 from neural crest cells leads tocorneal vascularization and anterior segment dysgenesisTsutomu Kume, Kathryn M. Schultz, Amy Sasman, Seungwoon Seo.Medicine, Northwestern Univ Sch of Med, Chicago, IL.Purpose: Foxc1 and Foxc2, members of the forkhead/Foxtranscription factor family, are critical regulators of vasculardevelopment. Foxc1 and Foxc2 are both expressed in ocularmesenchymes of neural crest origin. However, little is known aboutthe role of Foxc1 and Foxc2 in ocular development, mainly due to themid-gestation lethality of global knockout mice. Previously, we havedemonstrated that deletion of Foxc1 in neural crest-derived cells(NCC) leads to aberrant vessel growth in the normally avascularcorneas of mice, and that the effect is cell-type specific, because thecorneas of mice lacking Foxc1 expression in vascular endothelialcells remained avascular. Therefore, we investigate the role of Foxc1and Foxc2 in NCC during ocular development.Methods: To specifically delete Foxc1 and/or Foxc2 from NCC,conditional knockout mice for Foxc2 (NC-Foxc2-/-) and for Foxc1and Foxc2 (NC-Foxc1-/-; Foxc2-/-) were generated by crossingfloxed-Foxc1 and floxed-Foxc2 mice with Wnt1-cre mice.Results: Adult NCC-specific Foxc2 mutant mice exhibit corneal©2013, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permissionto reproduce any abstract, contact the ARVO Office at arvo@arvo.org.