Transcriptomic analysis of the role of Hdac4 in cardiac differentiation of mouse embryonic stem cells
Publication Date
July 16, 2024
Abstract
Embryonic heart development is a complex process that is intricately controlled by various transcription factors and signalling pathways. Perturbations to this process can lead to congenital heart disease, the most common cause of infant mortality in the UK. In this project, cellular models of heart development were used to investigate the function of histone deacetylation in cardiac differentiation. P19CL6 murine embryonic carcinoma cells and murine embryonic stem cells were differentiated and mESCs were chosen for downstream analyses. RNA-Seq was employed to study genes involved in normal differentiation, in addition to the consequences of nonspecific Class II HDACs inhibition using Trichostatin A (TSA) and Hdac4 knockout mediated by CRISPR/Cas9. Differentiated cells showed lower expression of pluripotency-associated genes such as Nanog and Sox2. Genes linked with processes like cytoskeletal assembly and extracellular matrix formation were upregulated. Results also suggested that Trichostatin A treatment and Hdac4 knockout may have contributed to changes in gene expression. TSA was found to alleviate repression of Tbx5 and Hand2. Hdac4 knockout uncovered a possible role for methylation in cardiomyocyte gene regulation. Contractility genes were also investigated by comparing the transcriptional profiles of beating with non-beating differentiated cells. Genes associated with congenital heart disease, such as Tnni3 and Myh6 were identified. This analysis also revealed a newly-identified pathogenic gene, Ryr2, as well as the candidate gene Sh3bgr. While the results are not fully conclusive and require further validation, this study corroborated that HDACs play a regulatory role in cardiac differentiation and contractility and found an interesting similarity between the consequences of pan-HDAC inhibition and specific Hdac4 knockout.
Item Type
ethesis
Thesis Type
PhD
Supervisors
Subjects (LC)
Associated Schools / Departments
School of Life Sciences
eprints ID
77192
UoN Repository URI
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