Corneal haze and neovascularization affect over 1.5 million Americans every year and are among the leading causes of blindness worldwide. Gene therapy is an attractive and novel approach to prevent/treat these corneal disorders. However, clinical utility of gene therapy is severely limited due to unavailability of tissue-targeted gene transfer methods. To develop tissue-targeted selective gene therapy approaches for the cornea, we hypothesized that foreign genes can be selectively expressed in the cornea at the desired site for selected duration using appropriate vectors and vector-delivery techniques. Preliminary in vivo experiments performed with AAV serotype 2 or 5, lentivirus or plasmid vectors containing CMV or hybrid CMV+chicken-?-actin promoter, and defined vector-delivery techniques demonstrated that transgene can be precisely expressed in keratocytes of normal/damaged corneas in vivo for the desired time period. Decorin (a small leucine-rich proteoglycan) gene therapy has been shown to prevent fibrosis and angiogenesis in various disease animal models. These reports led us to hypothesize that selective expression of decorin in keratocytes can inhibit/prevent corneal haze and neovascularization with minimal side effects. The in vitro studies performed to test this hypothesis demonstrated competence of decorin to inhibit keratocyte transformation to myofibroblasts. This transformation is known to cause corneal haze in vivo. We further hypothesize that selective tissue-targeted gene transfer approaches can be used to develop animal models for studying the specific function of disease-causing genes such as TGF? and BIGH3 in the adult cornea in vivo without altering their expression in vital organs. The function of such genes cannot be studied using conventional transgenic approaches because TGF?-deficient transgenic animals suffer lethal defects and die by 4 weeks of age.
The specific aims to test the hypotheses are 1) vector and vector-delivery techniques regulate level, duration, and location of transgene expression in keratocytes in vivo 2) decorin gene therapy can control corneal haze and 3) decorin gene therapy can inhibit corneal neovascularization. Using a mouse model, tested vectors, and optimized vector delivery-techniques, we will test Specific Aim 1 and will define short- and long-term selective gene transfer approaches for the cornea. Rabbit models will be used to test Specific Aims 2 and 3 by delivering decorin into keratocytes with optimal selective gene transfer methods.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
3R01EY017294-03S1
Application #
7845166
Study Section
Anterior Eye Disease Study Section (AED)
Program Officer
Shen, Grace L
Project Start
2007-04-01
Project End
2012-03-31
Budget Start
2009-04-01
Budget End
2010-03-31
Support Year
3
Fiscal Year
2009
Total Cost
$41,188
Indirect Cost
Name
University of Missouri-Columbia
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
153890272
City
Columbia
State
MO
Country
United States
Zip Code
65211
Gupta, Suneel; Fink, Michael K; Ghosh, Arkasubhra et al. (2018) Novel Combination BMP7 and HGF Gene Therapy Instigates Selective Myofibroblast Apoptosis and Reduces Corneal Haze In Vivo. Invest Ophthalmol Vis Sci 59:1045-1057
Anumanthan, Govindaraj; Wilson, Philip J; Tripathi, Ratnakar et al. (2018) Blockade of KCa3.1: A novel target to treat TGF-?1 induced conjunctival fibrosis. Exp Eye Res 167:140-144
Marlo, Todd L; Giuliano, Elizabeth A; Tripathi, Ratnakar et al. (2018) Altering equine corneal fibroblast differentiation through Smad gene transfer. Vet Ophthalmol 21:132-139
Anumanthan, Govindaraj; Gupta, Suneel; Fink, Michael K et al. (2018) KCa3.1 ion channel: A novel therapeutic target for corneal fibrosis. PLoS One 13:e0192145
Chaurasia, Shyam S; Lim, Rayne R; Parikh, Bhav H et al. (2018) The NLRP3 Inflammasome May Contribute to Pathologic Neovascularization in the Advanced Stages of Diabetic Retinopathy. Sci Rep 8:2847
Marlo, Todd L; Giuliano, Elizabeth A; Sharma, Ajay et al. (2017) Development of a novel ex vivo equine corneal model. Vet Ophthalmol 20:288-293
Gronkiewicz, Kristina M; Giuliano, Elizabeth A; Sharma, Ajay et al. (2016) Molecular mechanisms of suberoylanilide hydroxamic acid in the inhibition of TGF-?1-mediated canine corneal fibrosis. Vet Ophthalmol 19:480-487
Mohan, Rajiv R; Morgan, Brandie R; Anumanthan, Govindaraj et al. (2016) Characterization of Inhibitor of differentiation (Id) proteins in human cornea. Exp Eye Res 146:145-53
Gronkiewicz, K M; Giuliano, E A; Kuroki, K et al. (2016) Development of a novel in vivo corneal fibrosis model in the dog. Exp Eye Res 143:75-88
Sharma, Ajay; Anumanthan, Govindaraj; Reyes, Marcos et al. (2016) Epigenetic Modification Prevents Excessive Wound Healing and Scar Formation After Glaucoma Filtration Surgery. Invest Ophthalmol Vis Sci 57:3381-9

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