The Transcriptional Coactivator Peroxisome Proliferator–Activated Receptor (PPAR) Coactivator-1and the Nuclear Receptor PPAR Control the Expression of Glycerol Kinase and Metabolism Genes Independently of PPARActivation in Human White Adipocytes Anne Mazzucotelli, 1,2 Nathalie Viguerie, 1,2,3 Claire Tiraby, 1,2 Jean-Se´ bastien Annicotte, 4 Aline Mairal, 1,2 Eva Klimcakova, 1,2,3 Emmanuelle Lepin, 1,2 Paul Delmar, 5 Se´ bastien Dejean, 6 Genevie` ve Tavernier, 1,2 Corinne Lefort, 1,2 Juan Hidalgo, 7 Thierry Pineau, 8 Lluis Fajas, 4 Karine Cle´ ment, 9 and Dominique Langin 1,2,3,10 OBJECTIVE—The purpose of this work was to determine the pattern of genes regulated by peroxisome proliferator–activated receptor (PPAR) coactivator 1(PGC-1) in human adipocytes and the involvement of PPARand PPARin PGC-1transcrip- tional action. RESEARCH DESIGN AND METHODS—Primary cultures of human adipocytes were transduced with a PGC-1adenovirus and treated with PPARand PPARagonists. Variation in gene expres- sion was assessed using pangenomic microarrays and quantitative RT-PCR. To investigate glycerol kinase (GyK), a target of PGC-1, we measured enzymatic activity and glycerol incorporation into triglycerides. In vivo studies were performed on wild-type and PPAR / mice. The GyK promoter was studied using chromatin immunoprecipitation and promoter reporter gene assays. RESULTS—Among the large number of genes regulated by PGC-1independently of PPAR, new targets involved in metab- olism included the gene encoding GyK. The induction of GyK by PGC-1was observed at the levels of mRNA, enzymatic activity, and glycerol incorporation into triglycerides. PPARwas also upregulated by PGC-1. Its activation led to an increase in GyK expression and activity. PPARwas shown to bind and activate the GyK promoter. Experiments in mice confirmed the role of PGC-1and PPARin the regulation of GyK in vivo. CONCLUSIONS—This work uncovers novel pathways regu- lated by PGC-1and reveals that PPARcontrols gene expres- sion in human white adipocytes. The induction of GyK by PGC-1 and PPARmay promote a futile cycle of triglyceride hydrolysis and fatty acid reesterification. Diabetes 56:2467–2475, 2007 T he accumulation of white adipose tissue (WAT) predisposes to the development of an array of metabolic disturbances leading to type 2 diabe- tes and cardiovascular disease. In a search for new therapies, it has been postulated that targeting mo- lecular pathways that regulate thermogenesis may provide a plausible means of increasing energy expenditure (1). In that context, the opposite role of WAT specialized in energy storage in the form of triglycerides and brown adipose tissue (BAT) specialized in adaptive thermogene- sis is of great interest. In humans, BAT depots and brown adipocytes are sparsely distributed in the body and are not thought to contribute to a significant part of adaptive thermogenesis (2,3). Conversion of human white adipo- cytes into fat cells with some properties of brown adipo- cytes is an attractive therapeutic strategy (4). Stimulation of lipolysis in WAT without the concomitant use of re- leased fatty acids may be detrimental because fatty acids in excess will be deposited in other organs and may induce insulin resistance and cardiovascular complications. In- stead, simultaneous activation of lipolysis and fatty acid utilization within white adipocytes could allow a decrease or a stabilization of fat mass without systemic side effects. The transcriptional coactivator peroxisome prolifera- tor–activated receptor (PPAR) coactivator 1(PGC-1) was initially described as a metabolic regulator of adaptive thermogenesis in BAT (5). PGC-1cooperates with the From the 1 Institut National de la Sante´ et de la Recherche Me´ dicale (INSERM) U858, Obesity Research Laboratory, Toulouse, F-31432, France; 2 Paul Sabatier University, Louis Bugnard Institute, Institut Fe´de´ ratif de Recherche 31, Toulouse, F-31432, France; 3 INSERM, Franco-Czech Laboratory for Clinical Research on Obesity, Prague, CZ-10100, Czech Republic; 4 INSERM U834, Metabolism and Cancer Laboratory, Montpellier, F-34090, France; the 5 Math- e´ matiques Applique´es aux Syste´ mes Laboratory, Ecole Centrale Paris, Chat- enay Malabry, F-92295, France; the 6 Centre National de la Recherche Scientifique, Statistics and Probality Laboratory, Paul Sabatier University, Toulouse, F-31400, France; the 7 Institute of Neurosciences, Department of Cellular Biology, Physiology and Immunology, Faculty of Sciences, Autono- mous University of Barcelona, Barcelona, 08193, Spain; the 8 Institut National de la Recherche Agronomique, Pharmacology and Toxicology Laboratory, Toulouse, France; 9 INSERM U872, Human Research Center on Nutrition, Hoˆ tel Dieu, Paris, F-75181, France; and the 10 Centre Hospitalier Universitaire de Toulouse, Biochemistry Laboratory, Biology Institute of Purpan, Toulouse, F-31059, France. Address correspondence and reprint requests to Dominique Langin, INSERM U858, IFR31 Institute, BP 84225, 31432 Toulouse Cedex 4, France. E-mail: langin@toulouse.inserm.fr. Received for publication 18 October 2006 and accepted in revised form 8 July 2007. Published ahead of print at http://diabetes.diabetesjournals.org on 23 July 2007. DOI: 10.2337/db06-1465. Additional information for this article can be found in an online appendix at http://dx.doi.org/10.2337/db06-1465. BAT, brown adipose tissue; GFP, green fluorescent protein; GSEA, Gene Set Enrichment Analysis; GyK, glycerol kinase; PANTHER, Protein Analysis Through Evolutionary Relationships; PGC-1, peroxisome proliferator–acti- vated receptor coactivator 1; PPAR, peroxisome proliferator–activated receptor; PPRE, PPAR responsive element; ROS, reactive oxygen species; RXR, retinoic acid X receptor-; SLC25A4, solute carrier family 25 member 4; UCP, uncoupling protein; WAT, white adipose tissue. © 2007 by the American Diabetes Association. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. ORIGINAL ARTICLE DIABETES, VOL. 56, OCTOBER 2007 2467 Downloaded from http://diabetesjournals.org/diabetes/article-pdf/56/10/2467/385884/zdb01007002467.pdf by guest on 04 November 2022