The anterior surface of the eye functions as a barrier to the external environment and protects the delicate underlying structures from injury, in part, through the elaboration of the limbal and corneal epithelia. As self-renewing tissues, these epithelia are governed by stem cells, which play a crucial role in tissue homeostasis, regeneration, transplantation, gene therapy and in the pathogenesis of several anterior ocular surface diseases. Equally important for proper vision is the need for corneal transparency, which is achieved through avascularity. It is well-accepted that the limbal epithelium is the site f the corneal epithelial stem cells;however, major questions remain unresolved concerning how the limbal epithelium is regulated. Likewise, our understanding of factors that control angiogenesis is incomplete. microRNAS (miRNAs) are a major class of regulatory molecules that are part of the RNAi silencing machinery. While some studies have been directed towards deciphering the roles of miRNAs in the corneal epithelium, little is known about the miRNA signature in the stem cell-enriched limbal epithelium. We have recently discovered that miRs-103/107 are limbal-preferred. Furthermore, we have evidence that miRs-103/107 function to insure proper limbal epithelial cell-cell contact, autophagy and impact on cell cycle quiescence. Until recently, it was believed that miR-184, the most abundant corneal epithelial miRNA, functioned to attenuate miR-205, which insured proper cell migration and cell survival. We now have evidence that miR-184 may directly prevent corneal epithelial angiogenesis. Proper vision requires both a stabile limbal epithelial and corneal clarity;therefore we propose to focus on: (1 The roles of miRs-103/107 in assuring the integrity of the limbal epithelium and (2) how miR-184 functions to maintain corneal avascularity. To accomplish these goals, we will capitalize on our ability to elucidate miRNA target proteins and modulate miRNA and target protein levels in submerged cultures of human limbal and corneal epithelial keratinocytes and human microvascular endothelial cells. We will manipulate these cultured cells to form either 3-D organotypic rafts, or endothelial tubes, which mimic the in vivo tissues. We will assess the functional consequences of such miRNA and protein modulations with a combination of biochemical, molecular biological, cell biological and physiological approaches. By focusing on the biology of these miRNAs and their target proteins, our proposal represents a novel approach to understand: (i) how the limbal epithelium is regulated;and (ii) what contributes to the maintenance of corneal avascularity. Such knowledge has relevance to stem cell biology and clinically to ex vivo corneal epithelial transplantation. This proposal also has clinical implicatins beyond just corneal avascularity and may impact on pathological retinal angiogenesis. Ultimately our studies will provide rationales for the development of innovative treatment regimens focused on the use of either: (i) inhibitors of specific miRNAs or their targets;or (ii) delivery of miRNAs in patients with diseases that affect the ocular anterior segmental epithelia.

Public Health Relevance

Information from this project will impact on our understanding of how: (i) the stem cell-enriched limbal epithelium maintains proper cell-cell contacts, autophagy and cell cycle quiescence;properties that are essential for homeostasis;and (ii) the corneal epithelium assures avascularity. The contribution of stem cells to the wellbeing of the corneal epithelium is undeniable as these cells govern self-renewal and tissue regeneration processes. Transparency is essential for proper vision and requires an avascular cornea.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
2R01EY019463-05
Application #
8759983
Study Section
(DPVS)
Program Officer
Mckie, George Ann
Project Start
2009-04-01
Project End
2019-06-30
Budget Start
2014-08-01
Budget End
2015-06-30
Support Year
5
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Northwestern University at Chicago
Department
Dermatology
Type
Schools of Medicine
DUNS #
City
Chicago
State
IL
Country
United States
Zip Code
60611
Park, Jong Kook; Peng, Han; Yang, Wending et al. (2016) miR-184 exhibits angiostatic properties via regulation of Akt and VEGF signaling pathways. FASEB J :
Cipolla, Gabriel A; Park, Jong Kook; de Oliveira, Liana A et al. (2016) A 3'UTR polymorphism marks differential KLRG1 mRNA levels through disruption of a miR-584-5p binding site and associates with pemphigus foliaceus susceptibility. Biochim Biophys Acta 1859:1306-13
Park, Jong Kook; Peng, Han; Katsnelson, Julia et al. (2016) MicroRNAs-103/107 coordinately regulate macropinocytosis and autophagy. J Cell Biol 215:667-685
Park, Jong Kook; Yang, Wending; Katsnelson, Julia et al. (2015) MicroRNAs Enhance Keratinocyte Proliferative Capacity in a Stem Cell-Enriched Epithelium. PLoS One 10:e0134853
Peng, Han; Park, Jong Kook; Katsnelson, Julia et al. (2015) microRNA-103/107 Family Regulates Multiple Epithelial Stem Cell Characteristics. Stem Cells 33:1642-56
Peng, Han; Kaplan, Nihal; Yang, Wending et al. (2014) FIH-1 disrupts an LRRK1/EGFR complex to positively regulate keratinocyte migration. Am J Pathol 184:3262-71
Hamanaka, Robert B; Glasauer, Andrea; Hoover, Paul et al. (2013) Mitochondrial reactive oxygen species promote epidermal differentiation and hair follicle development. Sci Signal 6:ra8
Peng, Han; Katsnelson, Julia; Yang, Wending et al. (2013) FIH-1/c-kit signaling: a novel contributor to corneal epithelial glycogen metabolism. Invest Ophthalmol Vis Sci 54:2781-6
Peng, Han; Kaplan, Nihal; Hamanaka, Robert B et al. (2012) microRNA-31/factor-inhibiting hypoxia-inducible factor 1 nexus regulates keratinocyte differentiation. Proc Natl Acad Sci U S A 109:14030-4
Kaplan, Nihal; Fatima, Anees; Peng, Han et al. (2012) EphA2/Ephrin-A1 signaling complexes restrict corneal epithelial cell migration. Invest Ophthalmol Vis Sci 53:936-45

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