Targeted Gene Therapy and Nanomedicine Approaches to Treat Corneal Diseases

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Abstract Corneal fibrosis (scarring) is a 3rd leading cause of blindness in veterans, military personnel, and civilians. Globally, 23 million people have corneal opacity/fibrosis in one eye and 4.9 million people are blind in both eyes. Currently, >167,000 veterans are legally blind, >7000 veterans develop blindness each year and 2 million have impaired vision. The long-term goal of our research is to define efficient and safe therapies to treat corneal fibrosis in humans. In last 4 years of funding, we made many new discoveries and successfully advanced development including first time finding of MyoD (myoblast determination protein 1), IL-37 (interleukin 37), and autophagy-related genes in cornea and their significant role in corneal wound repair and fibrosis. Capitalizing on these, we propose testing three new hypotheses to advance the field of corneal wound healing and fibrosis treatment further. Our central hypothesis is that formation and elimination of myofibroblast in corneal stroma is a time-sensitive essential mechanism for scarless corneal repair in vivo and strategies targeting its timely resolution will provide non-surgical means of vision restoration. Currently, corneal transplantation is standard of care to restore vision and no-specific non-surgical therapies exist to cure corneal fibrosis without side effects in humans. This proposal tests three independent specific aims: Aim-1 tests a hypothesis that PEI2-GNP nanoparticle mediated localize and tissue-targeted MyoD gene silencing therapy will remove established corneal scars/fibrosis by facilitating de-differentiation of corneal myofibroblasts (CMFs) into fibroblast cells via an ERK/MAPK signaling in vivo in a rabbit model of wound healing. Aim-2 tests a hypothesis that dual-action IL-37 eye drops will inhibit corneal inflammation by limiting the availability of Caspase-1 and prevent corneal haze by binding to SMAD3 in vivo in an established rabbit CIT model that shows corneal inflammation and haze pathologies seen in patients. Aim-3 tests a hypothesis that open-field blast injury upsets corneal keratocytes’ homeostatic mechanism and wound healing properties in vivo via mTORC1-mediated dysfunctional autophagy through LC3I/II, Beclin, SQSTM1, LAMP1/2, TFEB, & vATPase proteins. Proposed studies will be accomplished employing established in vitro human and in vivo pre-clinical rabbit and rodent models of corneal fibrosis, actual open field blast site; clinical eye exams, multimodal 2D/3D eye imaging, histology, molecular and cellular assays, and transmission electron microscopy using our published method. Successful completion of project will advance the development of novel non-surgical strategies to treat corneal fibrosis, fill many knowledge gaps, and meet the mission of the Veterans Health Administration to provide best health care to our veterans.