Inhibition of Cyclin-dependent kinase 4 (CDK4) activity in adipocytes enhances their thermogenic program

PORTALES, A.; MIGUEL, I.; GIOVAMBATTISTA, A.

Mar del Plata

Congreso; LXI Reunión Científica Anual de la Sociedad Argentina de Investigación Clínica (SAIC); 2016

SAIC-SAI-SAFIS

Beige adipocytes are thermogenically competent cells that develop in white adipose tissue (WAT) depots in response to b-adrenergic receptors stimulation. Two general mechanisms are accepted for beige adipocytes generation: de novo adipogenesis from beige precursor cells, and transdifferentiation from white adipocytes. In previous studies we found that inhibition of cyclin-dependent kinase 4 (CDK4) activity in stromal vascular fraction cells (SVF) from WAT led to an increase in beige adipocyte markers. Here we wanted to assess if the inhibition of CDK4 activity is involved in white to beige adipocyte transdifferentiation. To this aim, SVF cells from the epididimal depot of C57BL/6J mice were isolated, cultured to confluence and differentiated with an adipogenic cocktail. On day 8 post-differentiation, cells were treated or not with an inhibitor of CDK4 (Palbociclib (PAL) 1μM; CTR) for 48hs. In additional experiments, after the inhibition period, CTR and PAL cells were treated with Forskolin (FSK) 10μM for 4hs in order to activate the thermogenic program (CTRF and PAL-F). Finally, on day 10 cells were processed for total RNA extraction and RT-qPCR quantification of thermogenic (Ucp1, Prdm16), beige (Cd137) and general adipogenic (adiponectin) markers. Results showed that inhibition of CDK4 in adipocytes resulted in a significant increase in Ucp1 and Prdm16 mRNA levels in the PALgroup compared to the CTR (p