Direct Conversion of Human Myoblasts into Brown-Like Adipocytes by Engineered Super-Active PPARγ

ObjectiveTo determine whether super-activation of PPAR can reprogram human myoblasts into brown-like adipocytes and to establish a new cell model for browning research. MethodsTo enhance the PPAR signaling, M3, the transactivation domain of MyoD, was fused to PPAR. PPAR and M3-PPAR-lentiviral vectors were used to convert human myoblasts into adipocytes. Brown adipocyte markers of the reprogrammed adipocytes were assessed by qPCR and protein analyses. White adipocytes differentiated from subcutaneous stromal vascular cells and perithyroid brown fat tissues were used as references. ResultsIn transient transfections, M3-PPAR had a stronger constitutive activity than PPAR by reporter assay. Although the transduction of either PPAR or M3-PPAR induced adipogenesis in myoblasts, M3-PPAR drastically induced the brown adipocyte markers of UCP1, CIDEA, and PRDM16 by 1,050, 2.4, and 5.0 fold, respectively and increased mitochondria contents by 4 fold, compared to PPAR. ConclusionsSuper-activation of PPAR can effectively convert human myoblasts into brown-like adipocytes and a new approach to derive brown-like adipocytes.