Novel Combination BMP7 and HGF Gene Therapy Instigates Selective Myofibroblast Apoptosis and Reduces Corneal Haze In Vivo.

Gupta, Suneel ORCID: 0000-0002-6141-3728, Fink, Michael K, Ghosh, Arkasubhra ORCID: 0000-0002-6570-5891, Tripathi, Ratnakar, Sinha, Prashant R, Sharma, Ajay, Hesemann, Nathan P, Chaurasia, Shyam S ORCID: 0000-0001-8725-676X, Giuliano, Elizabeth A and Mohan, Rajiv R (2018) Novel Combination BMP7 and HGF Gene Therapy Instigates Selective Myofibroblast Apoptosis and Reduces Corneal Haze In Vivo. Investigative ophthalmology & visual science, 59 (2). pp. 1045-1057.

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Official URL: http://dx.doi.org/10.1167/iovs.17-23308

Abstract

<h4>Purpose</h4>We tested the potential of bone morphogenic protein 7 (BMP7) and hepatocyte growth factor (HGF) combination gene therapy to treat preformed corneal fibrosis using established rabbit in vivo and human in vitro models.<h4>Methods</h4>Eighteen New Zealand White rabbits were used. Corneal fibrosis was produced by alkali injury. Twenty-four hours after scar formation, cornea received topically either balanced salt solution (BSS; n = 6), polyethylenimine-conjugated gold nanoparticle (PEI2-GNP)-naked plasmid (n = 6) or PEI2-GNP plasmids expressing BMP7 and HGF genes (n = 6). Donor human corneas were used to obtain primary human corneal fibroblasts and myofibroblasts for mechanistic studies. Gene therapy effects on corneal fibrosis and ocular safety were evaluated by slit-lamp microscope, stereo microscopes, quantitative real-time PCR, immunofluorescence, TUNEL, modified MacDonald-Shadduck scoring system, and Draize tests.<h4>Results</h4>PEI2-GNP-mediated BMP7+HGF gene therapy significantly decreased corneal fibrosis in live rabbits in vivo (Fantes scale was 0.6 in BMP7+HGF-treated eyes compared to 3.3 in -therapy group; P < 0.001). Corneas that received BMP7+HGF demonstrated significantly reduced mRNA levels of profibrotic genes: α-SMA (3.2-fold; P < 0.01), fibronectin (2.3-fold, P < 0.01), collagen I (2.1-fold, P < 0.01), collagen III (1.6-fold, P < 0.01), and collagen IV (1.9-fold, P < 0.01) compared to the -therapy corneas. Furthermore, BMP7+HGF-treated corneas showed significantly fewer myofibroblasts compared to the -therapy controls (83%; P < 0.001). The PEI2-GNP introduced >104 gene copies per microgram DNA of BMP7 and HGF genes. The recombinant HGF rendered apoptosis in corneal myofibroblasts but not in fibroblasts. Localized topical BMP7+HGF therapy showed no ocular toxicity.<h4>Conclusions</h4>Localized topical BMP7+HGF gene therapy treats corneal fibrosis and restores transparency in vivo mitigating excessive healing and rendering selective apoptosis in myofibroblasts.

Item Type:	Article
Uncontrolled Keywords:	Cornea, Animals, Rabbits, Corneal Opacity, Disease Models, Animal, Fibrosis, Gold, Polyethyleneimine, Hepatocyte Growth Factor, Drug Combinations, Tonometry, Ocular, In Situ Nick-End Labeling, Apoptosis, Intraocular Pressure, Plasmids, Female, Metal Nanoparticles, Bone Morphogenetic Protein 7, Myofibroblasts, Real-Time Polymerase Chain Reaction, Administration, Ophthalmic, Genetic Therapy
Depositing User:	Symplectic Admin
Date Deposited:	31 May 2019 10:01
Last Modified:	19 Jan 2023 00:42
DOI:	10.1167/iovs.17-23308
Open Access URL:	http://10.0.4.143/iovs.17-23308
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3043817