Corneal blindness affects millions of individuals world-wide and usually can only be treated by transplantation of donated tissue. In many regions of the world donor tissue is not available;in the US, the supply of corneas for transplant is threatened by increasing numbers of eyes with refractive surgeries. The long-term goal of this project is to address corneal blindness by developing therapies based on stem cells. This project has already identified and characterized adult human corneal stromal stem cells (CSSC), which can be expanded in culture while retaining the ability to adopt a keratocyte phenotype and to elaborate abundant stroma-like connective tissue. When CSSC were cultured on a scaffolding of aligned paralel nanofibers of poly(ester urethane)urea (PEUU), a biodegradable polymer, they secreted connective tissue with aligned collagen similar to that of normal corneal stroma. When CSSC were injected into stromas of lumican knockout mice, collagen in the stroma became more organized and corneal transparency was increased. Importantly, human CSSC exhibited an immune privilege in mice, remaining viable for months without immune rejection. The new proposal will test specific hypotheses emanating directly from these exciting findings.
Aim 1 documents the process of tissue regeneration by CSSC by testing the predictions that (a) CSSC first remove existing tissue by temporally regulated expression of a specific group of matrix metalloproteinases;(b) after this removal process, CSSC rebuild stromal tissue by deposition of new connective tissue;and (c) this regeneration process can remodel scar tissue similar to that causing most human corneal blindness.
Aim 2 combines CSSC with PEUU scaffolding to produce a novel bioengineered tissue and tests the predictions that (a) CSSC monolayers on PEUU nanofiber scaffolding can integrate and function as transparent stromal tissue in mouse corneal transplants;(b) CSSC can be incorporated into multilayered PEUU scaffolding with the thickness and strength of a corneal stroma;and (c) the multilayered constructs can functionally replace stroma after transplantation into rabbit corneas using deep anterior lamellar keratoplasty. The scientific impact of this study will be a demonstration of effectiveness of two novel stem-cell based therapeutic approaches for corneal scarring, indicating a readiness for translation of each to clinical trials.

Public Health Relevance

Corneal blindness is a health problem world-wide. For many individuals with this condition there is no option for restoration of vision. This project will use adult human stem cells to develop therapy for corneal scarring, a major source of blindness. These stem cells can be produced in large numbers, are safe, and do cause tissue rejection. Successful development of these therapies may open new opportunity to restore vision to large numbers of people.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY016415-07
Application #
8245697
Study Section
Anterior Eye Disease Study Section (AED)
Program Officer
Mckie, George Ann
Project Start
2005-04-01
Project End
2015-03-31
Budget Start
2012-04-01
Budget End
2013-03-31
Support Year
7
Fiscal Year
2012
Total Cost
$445,178
Indirect Cost
$151,331
Name
University of Pittsburgh
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Morgan, Siân R; Dooley, Erin P; Kamma-Lorger, Christina et al. (2016) Early wound healing of laser in situ keratomileusis-like flaps after treatment with human corneal stromal stem cells. J Cataract Refract Surg 42:302-9
Palchesko, Rachelle N; Funderburgh, James L; Feinberg, Adam W (2016) Engineered Basement Membranes for Regenerating the Corneal Endothelium. Adv Healthc Mater 5:2942-2950
Funderburgh, James L; Funderburgh, Martha L; Du, Yiqin (2016) Stem Cells in the Limbal Stroma. Ocul Surf 14:113-20
Syed-Picard, Fatima N; Du, Yiqin; Hertsenberg, Andrew J et al. (2016) Scaffold-free tissue engineering of functional corneal stromal tissue. J Tissue Eng Regen Med :
Romanowski, Eric G; Stella, Nicholas A; Brothers, Kimberly M et al. (2016) Predatory bacteria are nontoxic to the rabbit ocular surface. Sci Rep 6:30987
Hertsenberg, Andrew J; Funderburgh, James L (2016) Generation of Corneal Keratocytes from Human Embryonic Stem Cells. Methods Mol Biol 1341:285-94
Wang, Siran; Ghezzi, Chiara E; White, James D et al. (2015) Coculture of dorsal root ganglion neurons and differentiated human corneal stromal stem cells on silk-based scaffolds. J Biomed Mater Res A 103:3339-48
Syed-Picard, Fatima N; Du, Yiqin; Lathrop, Kira L et al. (2015) Dental pulp stem cells: a new cellular resource for corneal stromal regeneration. Stem Cells Transl Med 4:276-85
Massie, Isobel; Dziasko, Marc; Kureshi, Alvena et al. (2015) Advanced imaging and tissue engineering of the human limbal epithelial stem cell niche. Methods Mol Biol 1235:179-202
Kureshi, Alvena K; Funderburgh, James L; Daniels, Julie T (2014) Human corneal stromal stem cells exhibit survival capacity following isolation from stored organ-culture corneas. Invest Ophthalmol Vis Sci 55:7583-8

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