Howard, S;
(2014)
The regulation of Wt1 in the epicardium by the SWI/SNF-like BAF complex and thymosin β4.
Doctoral thesis , UCL (University College London).
Abstract
During development, cells of the epicardium undergo epithelial-mesenchymal transition (EMT) to generate multipotent progenitor cells (epicardium-derived cells, EPDCs) that contribute to multiple lineages of the cardiovascular system. Critical to this process is the epicardially-expressed Wilms’ tumour 1 (Wt1) gene. In the adult, the epicardium is largely quiescent and no longer expresses Wt1. Upon myocardial infarction (MI), however, the adult epicardium is reactivated; it re-expresses Wt1, reverts to its embryonic potency, and contributes to cardiac repair. Previous work showed that this reactivation is enhanced by ‘priming’ mice with the G-actin-binding protein thymosin β4 (Tβ4), although the molecular mechanisms were not defined. Here we investigated Wt1 regulation in the epicardium during development and post-MI and found Wt1 to be regulated by the SWI/SNF chromatin remodelling complex in combination with Tβ4. During development, Wt1-expressing cells in the epicardium expressed the SWI/SNF ATPases, Brg1 and Brm. Both Wt1 and SWI/SNF were largely absent from the uninjured adult epicardium, but the re-expression of Wt1 post-MI was accompanied by substantial upregulation of both Brg1 and Brm. Exogenous Tβ4 in the form of priming increased the post-MI expression of Wt1, Brg1 and Brm, whilst endogenous Tβ4 was found to be required for maximal Wt1 reactivation. Furthermore, we found that Tβ4 physically interacts with the SWI/SNF complex and that it potentiates SWI/SNF-mediated activation of specific regulatory regions of the Wt1 promoter and intron 1, suggesting that this interaction is functional. Loss of Brg1 in the epicardium using Gata5Cre;Brg1flox embryos showed that Brg1 is not required for Wt1 expression, which we postulate is due to compensation from Brm, whilst loss of Brg1 in Wt1-expressing lineages (using the tamoxifen-inducible Wt1CreERT2) caused no obvious phenotype. Further understanding of the mechanisms underlying Wt1 expression, during both development and epicardial reactivation post-MI, is essential in our quest to utilise resident-cell-based therapy in cardiac repair.
Type: | Thesis (Doctoral) |
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Title: | The regulation of Wt1 in the epicardium by the SWI/SNF-like BAF complex and thymosin β4 |
Language: | English |
UCL classification: | UCL > Provost and Vice Provost Offices UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health |
URI: | https://discovery.ucl.ac.uk/id/eprint/1419273 |
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