The role of Coup-TFII in the acquisition of cellular fates during the differentiat...
| Grant number: | 12/09602-0 |
| Support type: | Research Grants - Young Investigators Grants |
| Duration: | April 01, 2013 - March 31, 2017 |
| Field of knowledge: | Biological Sciences - Biochemistry |
| Principal Investigator: | Henrique Marques Barbosa de Souza |
| Grantee: | Henrique Marques Barbosa de Souza |
| Home Institution: | Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil |
| Assoc. researchers: | Alberto Sunao Stolfi ; José Xavier Neto ; Lucimara Aparecida Sensiate |
| Associated scholarship(s): | 16/13095-8 - Characterization of the relationship between the polarized expression of Coup-TFII and the Wnt/²-catenin pathway in embryoid bodies from murine embryonic stem cells,
BP.MS 16/03553-9 - Embryonic stem cells cultivation and differentiation in embryoid bodies, BP.IC 15/22067-5 - Analysis of gene expression in embryoid bodies, BP.IC 15/06732-9 - The role of Coup-TFII in the acquisition of cellular fates during the differentiation of embryonic stem cells, BP.MS |
Abstract
Gene regulatory Networks (GRN) controlled by transcription factors are the main molecular component acting on the maintenance or alteration of cell states. Among the GRNs described so far, the GRN controlling heart development is of great interest, due to its potential application for human health. The role of many genes and gene interactions have been characterized during cardiac differentiation, but very little is known about the genes controlling steps such as the patterning of the heart tube antero-posterior axis. This step is controlled by a Retinoic Acid gradient that, in short, will subdivide the heart tube into an anterior region, the future conus and ventricles, and a posterior region, that will become the sinoatrial compartment. The transcription factor Coup-TFII (chicken ovalbumin promoter-transcription factor II) has an important role in patterning the posterior region of the heart and is considered an important modulator of the Retinoic Acid signaling during heart development. As a transcription factor, Coup-TFII plays an important role in regulating directly, or indirectly, the expression of target genes. The aim of this project is to characterize the complete GRN controlled by Coup-TFII during heart development, using mice embryonic hearts as well as heart cells (cardiomyocytes) derived in vitro from mouse embryonic stem cells. Although many target genes have been described for Coup-TFII in different cellular contexts, this will be the first attempt to characterize all possible binding sites and target genes in a genome-wide context. To this aim, the cardiac cells expressing Coup-TFII will be processed for chromatin imunoprecipitation followed by deep sequencing (ChIP-seq). At first, this approach will reveal all the genomic regions where Coup-TFII bindings, revealing as well all the genes surrounding these regions as potential Coup-TFII target genes. With the use of loss-of-function strategies, such as gene silencing via RNAi, the expression of these potential target genes will be analyzed, revealing which one of these genes are indeed regulated directly by Coup-TFII. As a result, the genes that are misexpressed in the loss-of-function analysis but are not bound by Coup-TFII, will be considered indirect targets of Coup-TFII. (AU)