Activation of Wnt signaling using LY-2090314 in human limbal epithelial stem cell maintenance (Faith Ampadu)

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PROJECT SUMMARY Limbal epithelial stem/progenitor cells (LSCs) in the human corneal epithelium are the front line of defense to maintain transparency of the cornea. Loss or dysfunction of the LSCs leads to limbal stem cell deficiency (LSCD), which causes pain, inflammation, and loss of corneal transparency in patients. Cultivated LSCs from a patient’s healthy LSCs can be transplanted into the diseased cornea to replenish the LSCs, but maintaining sufficient LSCs and preventing LSC differentiation in culture to allow for a successful transplant remain a major challenge in the field. We have previously shown that activation of Wnt signaling using a Wnt mimic could enhance the LSC expansion in culture. Additionally, canonical and noncanonical Wnt activation operate in an integrated manner in the LSCs instead of a mutually exclusive manner as we previously thought. In this proposal, the effect of Wnt activation using a Wnt small molecule LY- 2090314 that inhibits GSK-3β will be investigated first using a TopFlash assay. The function and population of cultivated LSCs following LY-2090314 treatment in a dose dependent manner will be evaluated as described using biomarkers in the parent grant. The proposed project will further shed light on the mechanism of Wnt signaling in the regulation of LSCs. The results could inform further development of Wnt small molecules to maximize the population of undifferentiated LSCs in culture. The knowledge gained from the proposed project will allow continued improvement of patient-specific LSCD treatment.