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Principal Investigator: Panteleimon Rompolas
Organization: UNIVERSITY OF PENNSYLVANIA
Fiscal Year: 2024
Award: $732,045
Funding agency: National Eye Institute
PROJECT SUMMARY
Fuchs' endothelial corneal dystrophy (FECD) is a degenerative ocular disease affecting 4% of people over 40
and more that >300 million people globally. A hallmark of FECD is the accelerated loss of corneal endothelial
cells (CECs), which are required for maintaining corneal transparency and visual clarity. This study will
elucidate the underexplored regenerative potential of the corneal endothelium and use this knowledge to
devise innovative therapeutic interventions for FECD. Our hypothesis, grounded in preliminary experiments, is
that CECs, although naturally dormant, harbor an innate capacity for regeneration. Using innovative live
imaging approaches, genetic mouse models, and modified mRNA technology to manipulate CECs in vivo, we
identified a prospective therapeutic pathway: stimulating CEC proliferation in vivo to renew the corneal
endothelium, thus restoring corneal transparency and vision in FECD patients. To test this, we propose a two-
pronged approach utilizing relevant animal models. Aim 1 centers on elucidating the proliferative potential of
CECs in normal physiology and during FECD progression. Using established mouse models that recapitulate
the main features of FECD, we will perform real-time analysis of CEC dynamics by two-photon microscopy.
Moreover, we will investigate the molecular blueprint of a pro-regenerative endothelial cell identity, that can be
targeted for therapeutic intervention in FECD. For this we will perform in vivo assays of precision corneal
endothelial ablation combined with single-cell genomic analyses to elucidate the molecular programs that
influence the proliferation of CECs in vivo. To further illuminate the signaling pathways responsible for
regulating the cell cycle of CECs in response to changes of their biomechanical environment during FECD
disease progression, we will test the requirement of YAP, the downstream effector of the Hippo pathway, for
corneal endothelial function and regenerative ability. Aim 2 builds on our preliminary data demonstrating
effective control of CECs with injectable mRNA-encoded growth factors to induce their proliferation in vivo. We
propose a new mRNA-based therapy for FECD by directly targeting CECs to prompt the regeneration of a
diseased corneal endothelium, restoring its functionality. Toward this, we will systematically test the optimal
composition and delivery of our mRNA therapeutic factors to develop a robust treatment protocol. We will
comprehensively evaluate the effectiveness and safety of our mRNA treatment in two complementary mouse
models of early- and late-onset FECD, then scale up to pre-clinical trials in canine and non-human primate
models. Our team of investigators with deep expertise in regenerative biology and FECD disease modeling is
optimally equipped to carry out this research. With this study, we are aiming to not only revolutionize our
understanding of corneal endothelial physiology and FECD pathophysiology, but also develop groundbreaking
mRNA-based treatments to bring restored sight for patients with FECD and possibly other ocular degenerative
diseases closer to reality.
Terms: <2-photon microscopy><Ablation><Acceleration><Address><Affect><Aging><Animal Model><Animal Models and Related Studies><Antibodies><Assay><Behavior><Bioassay><Biological Assay><Biology><Biomechanics><Body Tissues><CRE Recombinase><Canine Species><Canis familiaris><Cell Body><Cell Cycle><Cell Density><Cell Division Cycle><Cell Growth in Number><Cell Multiplication><Cell Proliferation><Cell surface><Cells><Cellular Proliferation><Clinical><Complex><Cornea><Corneal Diseases><Corneal Disorder><Corneal Endothelium><Corneal dystrophy><Corneal edema><Data><Degenerative Disorder><Descemet's membrane><Disease><Disease Progression><Disorder><Dogs><Dogs Mammals><Dropsy><Dysfunction><Edema><Elderly><Endothelial Cells><Endothelium><Enterobacteria phage P1 Cre recombinase><Environment><Eye><Eye diseases><Eyeball><Fuchs endothelial corneal dystrophy><Fuchs' Endothelial Dystrophy><Fuchs' dystrophy><Functional disorder><Gene Expression><Gene variant><Genes><Genetic><Goals><Growth Agents><Growth Factor><Growth Substances><Human><Hydrops><Image><Impairment><Injectable><Injury><Investigators><Ion Pumps><Knowledge><Label><Lamina Elastica Posterior><Laser Electromagnetic><Laser Radiation><Lasers><Medicine><Messenger RNA><Methods><Mice><Mice Mammals><Mission><Modeling><Modern Man><Molecular><Monitor><Murine><Mus><National Eye Institute><Natural regeneration><Ocular Pathology><Pathogenesis><Pathway interactions><Patients><Persons><Phenotype><Physiology><Physiopathology><Proliferating><Proteins Growth Factors><Regeneration><Regenerative capacity><Regulation><Reporter><Research><Research Personnel><Researchers><Role><Safety><Sight><Signal Pathway><Specificity><Strategic Planning><Technology><Testing><Therapeutic><Therapeutic Intervention><Time><Tissues><Translating><Treatment Protocols><Treatment Regimen><Treatment Schedule><Vision><Visual><Wound Repair><absorption><advanced age><aged mice><aged mouse><allele variant><allelic variant><assess effectiveness><bacteriophage P1 recombinase Cre><biomechanical><canine><canine animal model><canine model><cornea disorder><cornea edema><corneal><corneal endothelial><degenerative condition><degenerative disease><determine effectiveness><disease model><disease phenotype><disorder model><dog model><domestic dog><dystrophia epithelialis corneae><early onset><effectiveness assessment><effectiveness evaluation><efficacy testing><elderly mice><endothelial regeneration><epigenomics><evaluate effectiveness><examine effectiveness><experiment><experimental research><experimental study><experiments><eye disorder><functional restoration><gain of function><genetic variant><genomic variant><geriatric><imaging><imaging approach><imaging based approach><in vivo><in vivo Model><injuries><innovate><innovation><innovative><intervention therapy><lipid based nanoparticle><lipid nanoparticle><loss of function><mRNA><migration><model of animal><mouse genetics><mouse model><murine model><new drug treatments><new drugs><new pharmacological therapeutic><new therapeutic approach><new therapeutic intervention><new therapeutic strategies><new therapeutics><new therapy><new therapy approaches><new treatment approach><new treatment strategy><next generation therapeutics><non-human primate><nonhuman primate><novel drug treatments><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel therapeutic approach><novel therapeutic intervention><novel therapeutic strategies><novel therapeutics><novel therapy><novel therapy approach><ocular disease><ocular disorder><old mice><ophthalmopathy><pathophysiology><pathway><pre-clinical development><pre-clinical trial><preclinical development><preclinical trial><progenitor cell population><progenitor population><programs><prospective><regenerate><regeneration ability><regeneration biology><regeneration capacity><regeneration potential><regenerative><regenerative approach><regenerative biology><regenerative potential><regenerative strategy><regenerative technique><response><restore function><restore functionality><restore lost function><restore sight><restore vision><scale up><senescence><senescent><senior citizen><sight restoration><single cell genomics><social role><stem and progenitor cell population><stem cell population><synergism><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><tool><two photon excitation microscopy><two photon microscopy><uptake><vision restoration><visual function><wound healing><wound recovery><wound resolution>