KD025 (SLx-2119) suppresses adipogenesis at intermediate stage in human adipose-derived stem cells

ABSTRACT

Rho-associated kinases (ROCKs) have been reported to antagonize adipocyte differentiation, and inhibition of ROCKs by small molecules promotes adipogenesis. Surprisingly, our recent study revealed that the ROCK2-specific inhibitor KD025 (SLx-2119), suppresses differentiation at the intermediate stage in 3T3-L1 preadipocytes. To address whether the anti-adipogenic activity of KD025 is a generalizable property, we examined the effect of KD025 in human adipose-derived stem cells (hADSCs). KD025 significantly suppressed the adipocyte differentiation of hADSCs with downregulation of the protein and mRNA expression of various adipogenic and lipogenic markers, including PPARγ, C/EBPα, SREBP-1c, Glut4 and FABP4. Notably, we observed that adipocyte differentiation is effectively suppressed by exposure to KD025 during the mid-to-late period of adipogenesis but not at the earlier stages, showing stage-specificity. Contrary to expectations, KD025 upregulated the insulin signaling, as confirmed by the increased phosphorylation levels of Akt and GSK-3α/β, and the differentiation-promoting activity of insulin signaling was observed to be overwhelmed by the inhibitory activity. In addition, we observed that other ROCK inhibitors (Y-27632, fasudil, and H-1152P) did not suppress but promoted adipocyte differentiation. These results indicate that KD025 suppresses adipocyte differentiation by modulation of key factors activated at the intermediate stage of differentiation, and not by inhibition of ROCK2.

KEYWORDS:

• KD025
• SLx-2119
• adipogenesis
• differentiation
• ROCK
• Rho-associated kinase

Disclosure statement

No potential conflict of interest was reported by the authors.

Author contributions

K.H.C. designed the experiments. D.T.V.D. performed the experiments. D.T.V.D., H.C. and K.H.C. analyzed the data and wrote the manuscript. K.H.M.D. and H.S.J. contributed to the editing of the manuscript. All authors reviewed the manuscript.

Additional Information

Competing financial interests: The authors declare no competing financial interests.

Additional information

Funding

This study was supported by grants from the Basic Science Research Program through the National Research Foundation of Korea, funded by the Ministry of Education [2016R1D1A1B01012515], Republic of Korea.