Characterizing the roles of transcription factor isoforms in rewiring gene regulatory networks during adipogenesis
Metabolism-related disorders like obesity are on the rise worldwide, posing a major global disease burden. Fat tissue is a key site of metabolic regulation, and understanding how adipocytes develop is an important step in designing effective treatment or intervention strategies. This proposal aims to understand the gene regulatory network (GRN) underlying adipogenesis and to which extent this GRN is modulated by transcription factor (TF) isoforms. Adipogenesis is regulated by a network of successively activated TFs. Many of these TFs have multiple isoforms, a handful of which (e.g. PPARγ) are known to function divergently from their reference forms. Thus, this project will investigate the potentially key role of TF isoforms in shaping the adipogenic program and its robustness. In Objective 1, I will combine computational analysis with a novel microfluidic in vitro TF–DNA binding assay to characterize the DNA binding specificities of TF isoforms with the goal of identifying those that have the potential to rewire GRNs. In Objective 2, I will screen for TF isoforms that modulate the process of adipogenesis, using a novel droplet-based single-cell technology to assay how adipocyte differentiation changes in response to TF isoform overexpression. ChIP-seq and single-cell transcriptome profiling, combined with powerful computational analysis, will elucidate the molecular basis of these changes in relation to GRNs. Findings from this project will improve our understanding of transcriptional mechanisms in adipogenesis. Furthermore, results will have broad implications for the role of TF isoforms in gene regulation, highlighting the complexity of gene regulatory mechanisms that underlie cell and tissue specificity.