Objective
To determine whether super-activation of PPARγ can reprogram human myoblasts into brownlike adipocytes and to establish a new cell model for browning research.
Methods
To enhance the PPARγ signaling, we fused M3, the transactivation domain of MyoD, 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 transfection, M3-PPARγ has stronger constitutive activity than PPARγ reporter assay. Although both the transduction of PPARγ and M3-PPARγ induced adipogenesis in myoblasts, M3-PPARγ, compared to 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. The gene expression levels of the browning makers in PPARγ-reprogrammed adipocytes are comparable to those of in vitro differentiated white adipocytes.
Conclusions
We have found that super-activation of PPARγ can effectively convert human myoblasts into brown-like adipocytes and can be a new approach to derive brown-like adipocytes.
Two genetic variants (rs3798220 and rs10455872) in the apolipoprotein (a) gene (LPA) have been implicated in cardiovascular disease (CVD), presumably through their association with lipoprotein (a) [Lp(a)] levels. While Lp(a) is recognized as a lipoprotein with atherogenic and thrombogenic characteristics, it is unclear whether or not the two Lp(a)-associated genetic variants are also associated with markers of thrombosis (i.e., plasminogen levels and fibrinolysis). In the present study, we genotyped the two genetic variants in 2919 subjects of the Old Order Amish (OOA) and recruited 146 subjects according to the carrier and noncarrier status for rs3798220 and rs10455872, and also matched for gender and age. We measured plasma Lp(a) and plasminogen levels in these subjects, and found that the concentrations of plasma Lp(a) were 2.62- and 1.73-fold higher in minor allele carriers of rs3798220 and rs10455872, respectively, compared with noncarriers (P = 2.04 × 10−17 and P = 1.64 × 10−6, respectively). By contrast, there was no difference in plasminogen concentrations between carriers and noncarriers of rs3798220 and rs10455872. Furthermore, we observed no association between carrier status of rs3798220 or rs10455872 with clot lysis time. Finally, plasminogen mRNA expression in liver samples derived from 76 Caucasian subjects was not significantly different between carriers and noncarriers of these two genetic variants. Our results provide further insight into the mechanism of action behind two genetic variants previously implicated in CVD risk and show that these polymorphisms are not major modulating factors for plasma plasminogen levels and fibrinolysis.
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