Novel Methods for Human Skeletal Muscle Regeneration Using Gene-Edited Pigs

Background, Rationale, Hypotheses and Objectives: Previous studies in this laboratory have defined the importance of immunologic tolerance to the field of transplantation. This project will examine the role of tolerance in enabling tissue complementation by humanized muscle produced in gene-edited animals.

Aims: The specific aims in this project will establish MHC inbred strains of miniature swine to produce specifically modified porcine embryos capable of being complemented by normal embryonic cells of the same or a different species, determine the extent of genetic disparities across which tolerance can be established, and determine whether immunologic tolerance in chimeric recipients is restricted to muscle cells or is directed to other cells and tissues. Tolerance will also be examined in the macaque-pig and human-pig chimeras.

Impact and Focus Area: The results of these studies should have both (1) theoretical implications for understanding tolerance induction following establishment of tissue chimerism at the embryonic level and (2) practical implications for development of a new technology for producing intra- and trans-species chimeras as a source of transplantable tissues. The ultimate goal will be to produce a reliable and reproducible source of skeletal muscle to treat muscle loss through reconstructive surgery, which directly addresses the FY19 JCP-8/CRMRP RMFRA Focus Area of Skeletal Muscle Regeneration.

Military Relevance: The availability of a reliable and reproducible source of skeletal muscle would enable treatment of VML at the time of battlefield injury or later, when muscle is required for restorative and reconstructive surgery. This technology would therefore have the potential to change the standard of care for battlefield injuries requiring such reconstructive care.