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Principal Investigator: Kate Tarvestad Laise
Organization: UNIVERSITY OF LOUISVILLE
Fiscal Year: 2024
Award: $35,822
Funding agency: National Eye Institute
Project Summary
The proposed research is relevant to public health because corneal blindness is the 4th leading cause of
blindness globally and is often preventable with proper healthcare. Blindness arises due to damaged integrity of
the corneal epithelium as the result of physical, chemical, or thermal trauma and/or as a consequence of
diseases, surgery, or anti-cancer drugs. Regardless of the cause, disruption of the epithelial layer is extremely
painful due to the dense sensory nerve innervation, makes the eye vulnerable to infection, and can result in
blindness. Despite the widespread incidence and the array of medical problems associated with epithelial
damage, there are no FDA-approved drugs that promote corneal wound healing or epithelial homeostasis. It is
well-established that proteins of the receptor tyrosine kinase (RTK) family can induce cell proliferation, migration,
survival, and differentiation of epithelial cells. We are investigating the signaling axis of one of these proteins, c-
Met, and its cognate ligand hepatocyte growth factor (HGF). c-Met activity can not only promote the wound
healing/re-epithelialization phenotypes stated previously but can also prevent transformation of keratocytes to
fibroblasts and promote neuronal growth. However, c-Met is limited for therapeutic reasons in that RTKs have
transient activation and are desensitized following ligand stimulation. We hypothesize that extending c-Met
receptor activity by disrupting its downregulation will help restore corneal homeostasis following
wounding. This hypothesis will be tested through the following aims: Aim 1: Determine if ubiquitylation of the c-
Met receptor limits its signaling in corneal epithelial cells in vitro. We hypothesize that inhibiting negative
regulation of c-Met signaling will prolong receptor activity and accelerate healing. We will test this by blocking c-
Met ubiquitylation in vitro by genetic knockout of CBL genes from immortalized human corneal epithelial cells
and measuring receptor phosphorylation, trafficking, in vitro wound healing, and cell migration and proliferation.
Aim 2: Determine effect of CBL knockout and c-Met activation in corneal wound healing murine models in vivo.
We hypothesize that HGF-induced c-Met signaling will not only aid in re-epithelialization, but also will decrease
fibrotic markers in the stromal layer and promote axonal outgrowth of corneal nerves. We will test this by using
corneal epithelial-specific, inducible CBL knockout murine models. We will wound the corneas and monitor gross
corneal morphology and homeostasis, re-epithelialization, fibrosis and inflammatory markers, and nerve
restoration. These studies are innovative because we will manipulate c-Met signaling regulation to extend
signaling and promote healing of all layers of the cornea. The proposed research is relevant to the National Eye
Institute’s mission of eliminating vision loss and improving quality of life through research.
Terms: <Acceleration><Anatomic Sites><Anatomic structures><Anatomy><Anterior><Anti-Cancer Agents><Antineoplastic Agents><Antineoplastic Drugs><Antineoplastics><Assay><Autoimmune Diseases><Autoregulation><Axon><Back><Binding><Bioassay><Biochemical><Biological Assay><Biology><Blindness><Body Tissues><Brain><Brain Nervous System><CBL><CBL gene><CBL2><Cancer Drug><Cas-Br-M (Murine) Ecotropic Retroviral Transforming Sequence><Cell Communication and Signaling><Cell Growth in Number><Cell Line><Cell Locomotion><Cell Migration><Cell Movement><Cell Multiplication><Cell Proliferation><Cell Signaling><CellLine><Cellular Migration><Cellular Motility><Cellular Proliferation><Chemicals><Cicatrix><Complication><Comprehension><Cornea><Corneal Injury><Corneal Opacity><Cranial Nerve V><Cytokine Receptors><Descemet's membrane><Diabetes Mellitus><Disease><Disorder><Dorsum><Down-Regulation><Drugs><Dryness><E3 Ligase><E3 Ubiquitin Ligase><Encephalon><Endothelium><Environment><Epithelial Cells><Epithelium><Eye><Eyeball><FDA approved><Family><Fibroblasts><Fibrosis><Fifth Cranial Nerve><Foreign Bodies><Generalized Growth><Genetic><Goals><Growth><Growth Agents><Growth Factor><Growth Substances><HGF gene><HGF/SF><Health><Healthcare><Hepatocyte Growth Factor><Hepatopoietin A><Homeostasis><Human><Human Figure><Human body><In Vitro><Incidence><Infection><Infectious Agent><Inflammation><Inflammation Mediators><Inflammatory><Intervention><Intervention Strategies><Intracellular Communication and Signaling><KO mice><Knock-out><Knock-out Mice><Knockout><Knockout Mice><Lamina Elastica Posterior><Leukoma><Ligands><Light><Lung Fibroblast-Derived Mitogen><Measures><Medical><Medication><Mission><Modeling><Modern Man><Molecular><Molecular Interaction><Monitor><Morphology><National Eye Institute><Neoplastic Disease Chemotherapeutic Agents><Nerve><Nerve Fibers><Nerve Regeneration><Nervus Trigeminus><Neuro-regeneration><Neuroregeneration><Null Mouse><Nutrient><O element><O2 element><Operative Procedures><Operative Surgical Procedures><Oxygen><PTK Receptors><Pain><Painful><Patients><Peripheral Nerves><Pharmaceutical Preparations><Pharmacodynamics><Phenotype><Phosphorylation><Photoradiation><Physiological Homeostasis><Physiology><Process><Proliferating><Protein Phosphorylation><Proteins><Proteins Growth Factors><Public Health><QOL><Quality of life><Receptor Protein><Receptor Protein-Tyrosine Kinases><Receptor Signaling><Receptor Tyrosine Kinase Gene><Recurrence><Recurrent><Regulation><Research><Research Support><Retina><Role><Scars><Scatter Factor><Sensory><Sight><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Strains Cell Lines><Structure><Surgical><Surgical Interventions><Surgical Procedure><System><Tamoxifen><Temperature><Testing><Therapeutic><Tissue Growth><Tissues><Transmembrane Receptor Protein Tyrosine Kinase><Transmission><Trauma><Trigeminal Nerve><Trigeminal nerve structure><Tumor-Specific Treatment Agents><Tyrosine Kinase Linked Receptors><Tyrosine Kinase Receptors><Ubiquitin Protein Ligase><Ubiquitin-Protein Ligase Complexes><Ubiquitin-Protein Ligase E3><Vision><Wound Repair><afferent nerve><anti-cancer drug><autoimmune condition><autoimmune disorder><autoimmunity disease><axon growth><axonal growth><biological signal transduction><cell motility><cornea opacity><corneal><corneal epithelial><corneal epithelial wound healing><corneal epithelium><corneal healing><corneal regeneration><corneal wound><corneal wound healing><cultured cell line><cytokine><desensitization><develop therapy><diabetes><drug/agent><epithelial injury><epithelium regeneration><expectation><healing><health care><improved><in vivo><in vivo Model><infectious organism><inflammation marker><inflammatory marker><inflammatory mediator><innervation><innovate><innovation><innovative><intervention development><interventional strategy><intraepithelial><migration><mouse model><murine model><nerve supply><nervous system regeneration><neural regeneration><neuronal growth><neuroregenerative><ontogeny><pathogen><pharmacologic><preservation><prevent><preventing><re-epithelialization><receptor><regenerate epithelium><regenerated nerve><response><restoration><sensory nerve><side effect><social role><surgery><therapy development><tissue wound><trafficking><transmission process><treatment development><ubiquitin-protein ligase><vision loss><visual function><visual loss><wound><wound healing><wound recovery><wound resolution><wounding><wounds>