Candidate: The candidate is a board-certified, cornea-trained ophthalmic surgeon with research interest in stem cell-base therapy for limbal stem cell deficiency. Her immediate career development goals in the current proposal will be learning in more details on cell biology, in vivo microscopy, image and data processing, and cell-based therapy. Her long-term career goals are to become a leader in the field of stem cell-based therapy of limbal stem cell deficiency and to develop into an independent clinician-scientist. The candidate plans an R01 submission in year 4 to establish an independent laboratory compatible with Good Laboratory Practice (GLP) and Good Tissue Practice (GTP) to evaluate the proposed therapy in phase I/II clinical trials. Additional didactic training in stem cll biology, in vivo microscopy, tissue engineering, and responsible conduct of research will be obtained during the award period. Environment: The mentorship of Drs. James Funderburgh and Robert Hendricks, and advisory committee combined with the degree of institutional commitment will provide this candidate with the support needed to transition successfully into an independent research career. Research: Prevalence of corneal blindness is second only to cataract worldwide. One of the clinically important causes of corneal blindness is limbal stem cell deficiency (LSCD). LSCD is caused by chemical, thermal, or autoimmune injury to the limbus area; results in persistent inflammation, vascularization and conjunctivalisation leading to vision loss. Despite advances in surgical techniques, LSCD remains one of the most difficult and challenging conditions for clinicians to manage. Conventional keratoplasty, keratoprosthesis and even keratolimbal auto- or allografts do not have decent long-term results. To date, there is a clear need for a cell source with ability of tissue regeneration and standardized method for treating LSCD. Human adipose derived stem cells (ADSCs) have multipotentiality, plasticity, immunomodulatory characteristics that make them attractive for use as an alternative therapy in LSCD. We have shown that ADSCs augment proliferation of human corneal epithelial cells. The long-term goal of the proposed research is to understand ADSC biology and develop a new treatment paradigm for limbal stem cell deficiency while allowing the PI to establish an independent research career in severe ocular surface disorders. The PI is well prepared to meet the scientific and training goals of this application through the research experience already acquired in the proposed research and mentorship environment, and continued development in this environment covered in this proposal. The overall hypothesis is that local administration of ADSCs will restore limbal niche environment and re-establish corneal epithelial integrity by anti- inflammatory/immunoregulatory mechanism. We will test the hypothesis with following specific aims:
Specific Aim 1. Determine the cellular and molecular interaction between human ADSCs and LESCs to preserve stemness of LESCs in a 3D in vitro co-culture system.
Specific Aim 2. Determine the dynamics of interaction between ADSCs and LESCs in the limbal area and demonstrate the therapeutic effect of TSG-6 secreted by ADSCs, in an animal model of LSCD.
Corneal diseases affect about 6-8 million people worldwide each year. A major cause of corneal blindness is loss of the stem cells in the limbal area, a situation known as limbal stem cell deficiency. My long-term goal is to develop into an independent clinician-scientist with a focus on ocular surface stem cell biology and regenerative therapy, with the hopes of developing alternative treatment regimens by using mesenchymal stem cells. If successful, the application will provide a basis for more effective therapies for the 750,000 Americans who each year suffers from acute and severe injuries to the ocular surface
