RFA-HL-23-019 RMIP II-Collaborative U01. Contact PI - Gary Yam Project Abstract Corneal blindness due to corneal scarring affects millions of people worldwide. Corneal transplantation is effective but has limitations. This Multi-PI project is aimed to establish a new stem cell-based approach to reduce the need for corneal allograft transplantation. The scopes of work in this project are to validate and confirm procedures to generate Good Manufacturing Practice (GMP) stem cells, verify the cell safety, and analyze clinical data towards a cell-based therapy for corneal scarring disorders. The Yam Lab and the Corneal Task Force at the University of Pittsburgh, following the innovative work by the late James L. Funderburgh, have confirmed the regenerative effects of human corneal stromal stem cells (CSSC) in animal models of corneal scarring. The Hsu Lab and the Immunologic Monitoring and Cellular Products Laboratory (IMCPL) at UPMC Hillman Cancer Center have immense experience in establishing Investigation New Drugs (IND) manufacturing procedures, quality assurance systems and documentation to support GMP cell product manufacturing and processing for FDA-registered clinical trials. This proposal is a joint force of the two expertise to pave a route to stem cell-based therapy in future clinical trials and clinical applications treating corneal scarring. Specific Aim 1 will establish Chemistry and Manufacturing Control (CMC) for GMP grade CSSC for allocation towards Investigation New Drugs (IND) in FDA. We will characterize and compare the quality of GMP raised CSSC and lab-generated cells, and establish a complete GMP production protocol. Specific Aim 2 will elucidate the pharmacological and toxicological effects of GMP-CSSC. We will study cell safety and efficacy using our established mouse corneal injury model and will determine CSSC distribution, metabolism, absorption, excretion, metabolism, and tumorigenicity. Specific Aim 3 will run clinical data mining and analysis that will direct future clinical trial design. We will do retrospective data analysis using corneal keratitis and scarring patient database at the UMPC Eye Center and obtain statistical data to direct future clinical trial design on patient eligibility and treatment outcome analysis. This project is established based on our solid basic and translational studies and data, and will lead to establish GMP-grade CSSC ready for clinical trials and to design clinical trials for treating corneal blindness.