oa Molecular Signature of Human Brown Adipocyte-Like PAZ6 Cells

Background: Prevalence of metabolic syndrome is directly correlated with increasing occurrence of obesity characterized by accumulation of fat in visceral, lower body, and upper body subcutaneous depots. In a number of species, brown adipose tissue (BAT) converts triglycerides into heat by non-shivering thermogenesis, thus controlling the amount of white adipose tissue. Until recently, BAT was thought to be mostly absent in adult humans but now sizeable BAT depots have been visualized by Positron Emission Tomography (PET) and Computed Tomography (CT) scanning. Relatively little is known about the signaling pathways differentially regulated in brown adipocytes compared to white adipocytes. However, our progress of understanding is hindered by the paucity of well-characterized reliable in vitro model systems.

Objectives: The aim of this study is to characterize the only in vitro model of human BAT cell line, and to identify signature genes for BAT, and compounds implicated in the enhancement of brown phenotype.

Methods: A human BAT cell line, PAZ6, was previously created by immortalizing somatic cells using the large T antigen of Simian Virus 40. The expression of BAT associated markers has been verified by real time RT-PCR done on RNA and immunoblot using specific antibodies performed on cell lysates and supernatants, before and after differentiation and treatment with various compounds such as adrenergic receptor agonist (isoproterenol) and PPAR agonist (rosiglitazone).

Results: We have identified several BAT associated markers, including PDRM16 and the 3 adrenergic receptor. This marker is highly expressed in PAZ6 cells compared to the white adipocyte. In addition, we demonstrate that these cells share a common precursor with myocytes but not with white adipocytes. Our results show that several genes are up-regulated after PPAR gamma activation. Moreover, we confirm that these cells respond to Beta adrenergic agonist treatment. Finally, we identify that the transcription factor farnesoid X Receptor (FXR) induces the expression of BAT associated marker following treatment by its agonist : CDCA.

Conclusions: The PAZ6 cells constitute a readily available surrogate for human brown adipocytes to develop pharmacologic strategies to promote BAT expansion and activation.