Transcriptomic Analysis of Cardiomyocyte Extracellular Vesicles in Hypertrophic Cardiomyopathy Reveals Differential snoRNA Cargo

James, Victoria; Nizamudeen, Zubair A.; Lea, Daniel; Dottorini, Tania; Holmes, Terri L.; Johnson, Ben B.; Arkill, Kenton P.; Denning, Chris; Smith, James G.W.

Abstract

Hypertrophic cardiomyopathy (HCM) is characterised by increased left ventricular wall thickness that can lead to devastating conditions such as heart failure and sudden cardiac death. Despite extensive study, the mechanisms mediating many of the associated clinical manifestations remain unknown and human models are required. To address this, human induced pluripotent stem cell (hiPSC) lines were generated from patients with a HCM associated mutation (c.ACTC1G301A) and isogenic controls created by correcting the mutation using CRISPR/Cas9 gene editing technology. Cardiomyocytes (hiPSC-CMs) were differentiated from these hiPSCs and analysed at baseline, and at increased contractile workload (2 Hz electrical stimulation). Released extracellular vesicles (EVs) were isolated and characterised following a 24 hour culture period and transcriptomic analysis performed on both hiPSC-CMs and released EVs. Transcriptomic analysis of cellular mRNA showed the HCM mutation caused differential splicing within known HCM pathways, and disrupted metabolic pathways. Analysis at increasing contraction frequency showed further disruption of metabolic gene expression, with an additive effect in the HCM background. Intriguingly, we observed differences in snoRNA cargo within HCM released EVs, that specifically altered when HCM hiPSC-CMs were subjected to increased workload. These snoRNAs were predicted to have roles in post translational modifications and alternative splicing, processes differentially regulated in HCM. As such, the snoRNAs identified in this study may unveil mechanistic insight into unexplained HCM phenotypes and offer potential future use as HCM biomarkers or as targets in future RNA-targeting therapies.

Citation

James, V., Nizamudeen, Z. A., Lea, D., Dottorini, T., Holmes, T. L., Johnson, B. B., …Smith, J. G. (2021). Transcriptomic Analysis of Cardiomyocyte Extracellular Vesicles in Hypertrophic Cardiomyopathy Reveals Differential snoRNA Cargo. Stem Cells and Development, 30(24), 1215-1227. https://doi.org/10.1089/scd.2021.0202

• Journal Article Type: Article
• Acceptance Date: Nov 3, 2021
• Online Publication Date: Nov 22, 2021
• Publication Date: Dec 15, 2021
• Deposit Date: Nov 8, 2021
• Publicly Available Date: Nov 23, 2022
• Journal: Stem Cells and Development
• Print ISSN: 1547-3287
• Electronic ISSN: 1557-8534
• Publisher: Mary Ann Liebert
• Peer Reviewed: Peer Reviewed
• Volume: 30
• Issue: 24
• Pages: 1215-1227
• DOI: https://doi.org/10.1089/scd.2021.0202
• Keywords: Cell Biology; Developmental Biology; Hematology
• Public URL: https://nottingham-repository.worktribe.com/output/6672967
• Publisher URL: https://www.liebertpub.com/doi/10.1089/scd.2021.0202
• Additional Information: Final publication is available from Mary Ann Liebert, Inc., publishers http://dx.doi.org/10.1089/scd.2021.0202