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, Benjamin B., Arkill, Kenton P., Denning, Chris and Smith, James G. W. (2021) Transcriptomic analysis of cardiomyocyte extracellular vesicles in hypertrophic cardiomyopathy reveals differential snoRNA cargo. Stem Cells and Development, 30 (24). pp. 1215-1227. ISSN 1547-3287

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Abstract

Hypertrophic cardiomyopathy (HCM) is characterized 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 analyzed at baseline, and at increased contractile workload (2 Hz electrical stimulation). Released extracellular vesicles (EVs) were isolated and characterized after a 24-h 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.

Item Type: Article
Additional Information: Funding Information: J.G.W.S. is supported by the Academy of Medical Sciences/ the Wellcome Trust/the Government Department of Business, Energy and Industrial Strategy/the British Heart Foundation/ Diabetes UK Springboard Award [SBF005\1057]. K.A is supported by the MRC [MR/P003214/1]; CD is funded by British Heart Foundation (SP/15/9/31605; RG/15/6/31436; 04BX14CDLG; PG/14/59/31000; RG/14/1/30588; P47352), National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs: NC/C013202/1; NC/ K000225/1).
Uncontrolled Keywords: cardiomyocytes,extracellular vesicles,hypertrophic,pluripotent stem cells,hematology,developmental biology,cell biology ,/dk/atira/pure/subjectarea/asjc/2700/2720
Faculty \ School: Faculty of Medicine and Health Sciences > Norwich Medical School
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Depositing User: LivePure Connector
Date Deposited: 07 Dec 2021 17:36
Last Modified: 24 May 2022 14:43
URI: https://ueaeprints.uea.ac.uk/id/eprint/82580
DOI: 10.1089/scd.2021.0202

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