Effect of deletion of the protein kinase PRKD1 on development of the mouse embryonic heart

Waheed‐Ullah, Qazi; Wilsdon, Anna; Abbad, Aseel; Rochette, Sophie; Bu'Lock, Frances; Hitz, Marc‐Phillip; Dombrowsky, Gregor; Cuello, Friederike; Brook, J. David; Loughna, Siobhan

Abstract

Congenital heart disease (CHD) is the most common congenital anomaly, with an overall incidence of approximately 1% in the United Kingdom. Exome sequencing in large CHD cohorts has been performed to provide insights into the genetic aetiology of CHD. This includes a study of 1891 probands by our group in collaboration with others, which identified three novel genes—CDK13, PRKD1, and CHD4, in patients with syndromic CHD. PRKD1 encodes a serine/threonine protein kinase, which is important in a variety of fundamental cellular functions. Individuals with a heterozygous mutation in PRKD1 may have facial dysmorphism, ectodermal dysplasia and may have CHDs such as pulmonary stenosis, atrioventricular septal defects, coarctation of the aorta and bicuspid aortic valve. To obtain a greater appreciation for the role that this essential protein kinase plays in cardiogenesis and CHD, we have analysed a Prkd1 transgenic mouse model (Prkd1 em1 ) carrying deletion of exon 2, causing loss of function. High‐resolution episcopic microscopy affords detailed morphological 3D analysis of the developing heart and provides evidence for an essential role of Prkd1 in both normal cardiac development and CHD. We show that homozygous deletion of Prkd1 is associated with complex forms of CHD such as atrioventricular septal defects, and bicuspid aortic and pulmonary valves, and is lethal. Even in heterozygotes, cardiac differences occur. However, given that 97% of Prkd1 heterozygous mice display normal heart development, it is likely that one normal allele is sufficient, with the defects seen most likely to represent sporadic events. Moreover, mRNA and protein expression levels were investigated by RT‐qPCR and western immunoblotting, respectively. A significant reduction in Prkd1 mRNA levels was seen in homozygotes, but not heterozygotes, compared to WT littermates. While a trend towards lower PRKD1 protein expression was seen in the heterozygotes, the difference was only significant in the homozygotes. There was no compensation by the related Prkd2 and Prkd3 at transcript level, as evidenced by RT‐qPCR. Overall, we demonstrate a vital role of Prkd1 in heart development and the aetiology of CHD.

• Journal Article Type: Article
• Acceptance Date: Feb 15, 2024
• Online Publication Date: Feb 28, 2024
• Publication Date: 2024-07
• Deposit Date: Feb 29, 2024
• Publicly Available Date: Feb 29, 2024
• Journal: Journal of Anatomy
• Print ISSN: 0021-8782
• Electronic ISSN: 1469-7580
• Publisher: Wiley
• Peer Reviewed: Peer Reviewed
• Volume: 245
• Issue: 1
• Pages: 70-83
• DOI: https://doi.org/10.1111/joa.14033
• Keywords: high‐resolution episcopic microscopy, Prkd1, protein kinase D1, congenital heart disease, protein kinase
• Public URL: https://nottingham-repository.worktribe.com/output/31893183
• Publisher URL: https://onlinelibrary.wiley.com/doi/10.1111/joa.14033