Direct conversion of human myoblasts into brown-like adipocytes by engineered super-active PPARγ.
OBJECTIVE
To determine whether super-activation of PPARγ can reprogram human myoblasts into brown-like adipocytes and to establish a new cell model for browning research.


METHODS
To enhance the PPARγ signaling, M3, the transactivation domain of MyoD, was fused to PPARγ. PPARγ and M3-PPARγ-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.


RESULTS
In transient transfections, M3-PPARγ had a stronger constitutive activity than PPARγ by reporter assay. Although the transduction of either PPARγ or M3-PPARγ induced adipogenesis in myoblasts, M3-PPARγ 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 PPARγ.


CONCLUSIONS
Super-activation of PPARγ can effectively convert human myoblasts into brown-like adipocytes and a new approach to derive brown-like adipocytes.
Medical School of Nanjing University, Nanjing, China; Division of Endocrinology, Diabetes and Nutrition, Department of Medicine, University of Maryland School of Medicine, Baltimore, USA.
Yanbei
Y
Zhu
Rongze
R
Yang
John
J
McLenithan
Daozhan
D
Yu
Hong
H
Wang
Yaping
Y
Wang
Devinder
D
Singh
John
J
Olson
Carole
C
Sztalryd
Dalong
D
Zhu
Da-Wei
DW
Gong
D017667:Adipocytes / Q000378:metabolism*; D052437:Adipocytes, Brown / Q000378:metabolism*; D052438:Adipocytes, White / Q000378:metabolism*; D050156:Adipogenesis / Q000502:physiology; D002001:Adipose Tissue, Brown / Q000378:metabolism*; D002454:Cell Differentiation / Q000502:physiology; D006801:Humans; D008928:Mitochondria / Q000378:metabolism; D032446:Myoblasts / Q000378:metabolism; D047495:PPAR gamma / Q000378:metabolism*; D017154:Stromal Cells / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism
D047495:PPAR gamma; D014157:Transcription Factors