Feasibility of Direct Reprogramming of Myofibroblasts to Lung Epithelial Cells Utilizing PiggyBac Transposon System

Organ transplantation is the treatment of choice for management of end-stage organ diseases. Graft failure due to chronic rejection is characterized by fibrosis of structures within the allograft. Immunosuppression has reduced acute rejection but has not had an impact on chronic rejection. In transplanted lungs, epithelial injury predestines small airways to undergo fibrosis and results in activation of myofibroblasts marked by expression of alpha smooth muscle actin (SMA) termed Bronchiolitis Obliterans (BO). In this project we developed an innovative anti-fibrotic strategy using "direct reprogramming" to target this pathogenic response through conversion of myofibroblast to lung epithelial cells. As a proof of concept, we 1) established an in vitro fibrosis model system, 2) tested SMA promoter as a potential driving promoter of the reprogramming cassette, 3) selected and cloned inducing factors in PiggyBac constructs and 4) showed the feasibility of utilizing these tools for the testing of reprogramming strategies targeting fibrosis.