Acute treatment with chemical PPARGC1a1 activators in brown fat cells

The peroxisome proliferator-activated receptor-coactivator-1α1 (PGC-1α1) regulates genes involved in energy metabolism. Increasing adipose tissue energy expenditure through PGC-1α1 activation has been suggested to be beneficial for systemic metabolism. Pharmacological PGC-1α1 activators could be valuable tools in the fight against obesity and metabolic disease. Finding such compounds has been challenging partly because PGC-1α1 is a transcriptional coactivator with no known ligand-binding activities. Importantly, PGC-1α1 activation is regulated by several mechanisms but protein stabilization is a limiting step as the protein has a short half-life under unstimulated conditions. We designed a cell-based high-throughput system to identify stabilizers of PGC-1α1 protein. Positive hits were tested for their ability to induce endogenous PGC-1α1 protein accumulation and activate target gene expression in brown adipocytes. Selected compounds were analyzed for their effects on global gene expression in brown adipocytes. Fully differentiated brown fat cells (previously described Uldry M et al 2006 Cell Metab.; PMID16679291) were treated for 8h with 10μM of compound (AM73 or AM80) or DMSO (control), in triplicate. RNA was extracted and used for global gene expression analyzes.