The immediate objective of this application is the isolation and characterization of stem cells in the closely related limbo-corneal and conjunctive epithelial lineages. The new knowledge from these studies will provide the fundamental information needed to develop strategies for clinical reconstruction of corneal tissues. Recent studies indicate that a large variety of somatic stem cells express the multidrug resistance transporter (MDR) ATP binding Cassette (ABC) transporter, ABCG2/BCRP. The activity of this transporter causes ceils to appear in flow cytometry blue vs. red fluorescence emission plots as a characteristically isolated group of cells positioned to the left (blue) side of the normal blue/red ratio. These cells have been termed side population (SP) cells. Accordingly both the ABCG2 + and SP phenotypes have been proposed to be general features of stem cells. We have recently identified ABCG2 in the human limbal and conjunctival epithelia and isolated SP cells from these tissues in human and rabbit by fluorescence activated cell sorter. The isolated ceils display multiple features associated with the epithelial stem cell phenotype including, a) an extremely low intracellular 'granularity' and a small size; b) a quiescent phenotype in vivo; c) a selective resiliency to induced differentiation by phorbol esters in vitro; and d) slow recruitment to proliferation when set in culture. We propose now to characterize these cells in detail from several biological and biochemical perspectives. We will a) examine whether SP or ABCG2 + cells exhibit the most recognizable features of epithelial stem cells including slow cycling and long term proliferative capacity; b) determine the complexity and diversity of the stem/precursor cell population; and c) identify ocular surface SP cell associated genes. Completion of these initial studies will provide critical new information on the features of the ocular precursor cell pool.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY014878-03
Application #
7176762
Study Section
Special Emphasis Panel (ZRG1-AED (01))
Program Officer
Shen, Grace L
Project Start
2005-01-15
Project End
2009-12-31
Budget Start
2007-01-01
Budget End
2007-12-31
Support Year
3
Fiscal Year
2007
Total Cost
$411,475
Indirect Cost
Name
Mount Sinai School of Medicine
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
078861598
City
New York
State
NY
Country
United States
Zip Code
10029
Wolosin, J Mario; Ritch, Robert; Bernstein, Audrey M (2018) Is Autophagy Dysfunction a Key to Exfoliation Glaucoma? J Glaucoma 27:197-201
Bernstein, Audrey M; Ritch, Robert; Wolosin, Jose M (2018) Exfoliation Syndrome: A Disease of Autophagy and LOXL1 Proteopathy. J Glaucoma 27 Suppl 1:S44-S53
Lee, Hyun Jung; Wolosin, J Mario; Chung, So-Hyang (2017) Divergent effects of Wnt/?-catenin signaling modifiers on the preservation of human limbal epithelial progenitors according to culture condition. Sci Rep 7:15241
Wolosin, J Mario; Zamudio, Aldo; Wang, Zheng (2017) Application of JC1 for non-toxic isolation of cells with MDR transporter activity by flow cytometry. PLoS One 12:e0174905
Selver, Ozlem Barut; Durak, Ismet; G├╝rdal, Mehmet et al. (2016) Corneal recovery in a rabbit limbal stem cell deficiency model by autologous grafts of tertiary outgrowths from cultivated limbal biopsy explants. Mol Vis 22:138-49
Zamudio, Aldo; Wang, Zheng; Chung, So-Hyang et al. (2016) Inhibition of TGF? cell signaling for limbal explant culture in serumless, defined xeno-free conditions. Exp Eye Res 145:48-57
Want, Andrew; Gillespie, Stephanie R; Wang, Zheng et al. (2016) Autophagy and Mitochondrial Dysfunction in Tenon Fibroblasts from Exfoliation Glaucoma Patients. PLoS One 11:e0157404
Yang, Yuanquan; Wang, Zheng; Yang, Hua et al. (2013) TRPV1 potentiates TGF?-induction of corneal myofibroblast development through an oxidative stress-mediated p38-SMAD2 signaling loop. PLoS One 8:e77300
Jeon, Sohee; Choi, Seong Hyun; Wolosin, J Mario et al. (2013) Regeneration of the corneal epithelium with conjunctival epithelial equivalents generated in serum- and feeder-cell-free media. Mol Vis 19:2542-50
Yang, Yuanquan; Yang, Hua; Wang, Zheng et al. (2013) Functional TRPV1 expression in human corneal fibroblasts. Exp Eye Res 107:121-9

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