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Suneel Gupta, Nishant Rajiv Sinha, Lynn M Martin, Landon Keele, Nathan P. Hesemann, Prashant Sinha, Elizabeth A Giuliano, Ratnakar Tripathi, Rajiv R Mohan; Evaluations of localized topical tissue-targeted AAV5-Id3 gene delivery to treat. Invest. Ophthalmol. Vis. Sci. 2022;63(7):117 – A0215.
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© ARVO (1962-2015); The Authors (2016-present)
Inhibitor of differentiation 3 (Id3), a transcriptional repressor gene, over-expression effectively inhibited differentiation of corneal fibroblast to myofibroblast in vitro. This study examined the therapeutic potential of targeted AAV5-Id3 gene delivery into stroma to treat corneal fibrosis using an established rabbit model.
The study was approved by the institutional animal care and use committee and followed ARVO guidelines. Eighteen New Zealand White rabbits were divided into 3 groups. Only one eye of each rabbit was used. Eyes of group-1 (naïve) received vehicle (n=6); group-2 received AAV5-naked following alkali injury, (n-=6); and group-3 received AAV5-Id3 following alkali injury (n=6). Gene vector into stroma was introduced using an optimized reported topical method. Eyes were periodically examined clinically with the slit lamp microscope, stereomicroscope, pachymetry, tonometry, and in vivo confocal microscopy. Fantes and Modified Hackett-McDonald scores were used to record clinical findings. At the end of 4-weeks, rabbits were humanely euthanized, and corneas were collected for histological, immunofluorescence, and qRT-PCR analyses.
Multimodal clinical imaging and ocular eye examinations revealed that AAV5-Id3 gene delivery significantly reduced corneal fibrosis compared to the AAV5-naked group (p < 0.01) without significant ocular manifestations. The qRT-PCR data indicated a significant reduction in mRNA levels of profibrotic genes; αSMA, fibronectin, collagen I, and collagen III (1.4-to-3.2-fold; p<0.01 or p<0.001) in eyes delivered AAV5-Id3 as compared to the AAV5-naked vector group. Histological and immunofluorescence data (H&E, Cd11b, and α-SMA staining) showed noticeably reduced inflammation and pro-fibrotic proteins levels in AAV5-Id3 therapy group compared to the AAV5-naked vector group. Histological and molecular data corresponds well with masked clinical exams.
Targeted AAV5-Id3 gene transfer into stroma effectively reduces corneal fibrosis in vivo in rabbits without gross clinical toxicity.
This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.
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