FatTFIso · Characterizing the roles of transcription factor isoforms in rewiring gene regulatory networks during adipogenesis
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2020-03-01 → 2022-02-28
- EU contribution
- €203,149
- Participants
- 1
- Scheme
- MSCA-IF
Lines connect the coordinator with its partners.
Results in brief
Characterizing the roles of transcription factor isoforms in rewiring gene regulatory networks during adipogenesis
Metabolism-related disorders pose a major global disease burden, as the rate of obesity rises worldwide. Fat tissue is a key site of metabolic regulation, and understanding how fat cells (adipocytes) develop forms the foundation for designing effective therapeutic and intervention strategies. This proposal aimed to understand the gene regulatory network (GRN) underlying fat cell development (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 exist as multiple isoforms, a handful of which are known to function divergently from their reference forms. Most functional characterization studies have been restricted to gene-level studies, largely ignoring isoforms. Because TFs function at critical steps in adipogenesis, such functionally different isoforms could determine the difference between cells that differentiate into mature fat cells and those that remain in the precursor states. This project aimed to investigate the extent to which TF isoforms are functionally different in the context of adipogenesis by conducting two screens: an in vitro screen for TF isoforms demonstrating different DNA binding specificities using a microfluidics-based assay, and an in vivo screen for isoforms that alter adipocyte differentiation using a novel droplet-based single-cell technology assay. Findings from this project were expected to improve our understanding of transcriptional mechanisms in adipogenesis and have broad implications for the role of TF isoforms in gene regulation.
Data: CORDIS, © European Union
Project objective
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.
Original text from CORDIS.
Participants
- ECOLE POLYTECHNIQUE FEDERALE DE LAUSANNE · LausanneCoordinatorSwitzerland
Links
Data: CORDIS, © European Union