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

Abstract

Purpose:

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.

Methods:

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.

Results:

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.

Conclusions:

Localized topical BMP7+HGF gene therapy treats corneal fibrosis and restores transparency in vivo mitigating excessive healing and rendering selective apoptosis in myofibroblasts.

• Authors: Suneel Gupta, Michael K Fink, Arkasubhra Ghosh, Ratnakar Tripathi, Prashant R Sinha, Ajay Sharma, Nathan P Hesemann, Shyam S Chaurasia, Elizabeth A Giuliano, Rajiv R Mohan
• Journal: Investigative ophthalmology & visual science (Invest Ophthalmol Vis Sci) Vol. 59 Issue 2 Pg. 1045-1057 (02 01 2018) ISSN: 1552-5783 [Electronic] United States
• PMID: 29490341 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• Bone Morphogenetic Protein 7
• Drug Combinations
• Hepatocyte Growth Factor
• Gold
• Polyethyleneimine

Topics

• Administration, Ophthalmic
• Animals
• Apoptosis (drug effects)
• Bone Morphogenetic Protein 7 (genetics)
• Cornea (pathology)
• Corneal Opacity (pathology, therapy)
• Disease Models, Animal
• Drug Combinations
• Female
• Fibrosis (therapy)
• Genetic Therapy (methods)
• Gold (chemistry)
• Hepatocyte Growth Factor (genetics)
• In Situ Nick-End Labeling
• Intraocular Pressure
• Metal Nanoparticles (chemistry)
• Myofibroblasts (pathology)
• Plasmids (genetics)
• Polyethyleneimine (chemistry)
• Rabbits
• Real-Time Polymerase Chain Reaction
• Tonometry, Ocular