Characterisation of the developing heart in a pressure overloaded model utilising RNA sequencing to direct functional analysis

Parnall, Matthew; Perdios, Chrysostomos; Pang, Kar Lai; Rochette, Sophie; Loughna, Siobhan

Abstract

Cardiogenesis is influenced by both environmental and genetic factors, with blood flow playing a critical role in cardiac remodelling. Perturbation of any of these factors could lead to abnormal heart development and hence the formation of congenital heart defects. Although abnormal blood flow has been associated with a number of heart defects, the effects of abnormal pressure load on the developing heart gene expression profile have to date not clearly been defined. To determine the heart transcriptional response to haemodynamic alteration during development, outflow tract (OFT) banding was employed in the chick embryo at Hamburger and Hamilton stage (HH) 21. Stereological and expression studies, including the use of global expression analysis by RNA sequencing with an optimised procedure for effective globin depletion, were subsequently performed on HH29 OFT‐banded hearts and compared with sham control hearts, with further targeted expression investigations at HH35. The OFT‐banded hearts were found to have an abnormal morphology with a rounded appearance and left‐sided dilation in comparison with controls. Internal analysis showed they typically had a ventricular septal defect and reductions in the myocardial wall and trabeculae, with an increase in the lumen on the left side of the heart. There was also a significant reduction in apoptosis. The differentially expressed genes were found to be predominately involved in contraction, metabolism, apoptosis and neural development, suggesting a cardioprotective mechanism had been induced. Therefore, altered haemodynamics during development leads to left‐sided dilation and differential expression of genes that may be associated with stress and maintaining cardiac output.

Citation

Parnall, M., Perdios, C., Pang, K. L., Rochette, S., & Loughna, S. (2020). Characterisation of the developing heart in a pressure overloaded model utilising RNA sequencing to direct functional analysis. Journal of Anatomy, 236(3), 549-563. https://doi.org/10.1111/joa.13112

• Journal Article Type: Article
• Acceptance Date: Oct 7, 2019
• Online Publication Date: Nov 14, 2019
• Publication Date: 2020-03
• Deposit Date: Oct 24, 2019
• Publicly Available Date: Nov 15, 2019
• Journal: Journal of Anatomy
• Print ISSN: 0021-8782
• Electronic ISSN: 1469-7580
• Publisher: Wiley
• Peer Reviewed: Peer Reviewed
• Volume: 236
• Issue: 3
• Pages: 549-563
• DOI: https://doi.org/10.1111/joa.13112
• Keywords: Haemodynamics, heart development, chick embryo, RNA sequencing, differential gene expression
• Public URL: https://nottingham-repository.worktribe.com/output/2958010
• Publisher URL: https://onlinelibrary.wiley.com/doi/10.1111/joa.13112
• Contract Date: Oct 24, 2019