Inducible apelin receptor knockdown reduces differentiation efficiency and contractility of hESC-derived cardiomyocytes.
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Authors
Colzani, Maria T
Williams, Thomas L
Bayraktar, Semih
Kuc, Rhoda E
Bernard, William G
Davenport, Emma E
Maguire, Janet J
Sinha, Sanjay
Davenport, Anthony P
Publication Date
2022-05-16Journal Title
Cardiovasc Res
ISSN
0008-6363
Publisher
Oxford University Press (OUP)
Type
Article
This Version
VoR
Metadata
Show full item recordCitation
Macrae, R. G., Colzani, M. T., Williams, T. L., Bayraktar, S., Kuc, R. E., Pullinger, A. L., Bernard, W. G., et al. (2022). Inducible apelin receptor knockdown reduces differentiation efficiency and contractility of hESC-derived cardiomyocytes.. Cardiovasc Res https://doi.org/10.1093/cvr/cvac065
Abstract
AIMS: The apelin receptor, a G protein-coupled receptor, has emerged as a key regulator of cardiovascular development, physiology, and disease. However, there is a lack of suitable human in vitro models to investigate the apelinergic system in cardiovascular cell types. For the first time we have used human embryonic stem cell-derived cardiomyocytes (hESC-CMs) and a novel inducible knockdown system to examine the role of the apelin receptor in both cardiomyocyte development and to determine the consequences of loss of apelin receptor function as a model of disease. METHODS AND RESULTS: Expression of the apelin receptor and its ligands in hESCs and hESC-CMs was determined. hESCs carrying a tetracycline-inducible short hairpin RNA targeting the apelin receptor were generated using the sOPTiKD system. Phenotypic assays characterized the consequences of either apelin receptor knockdown before hESC-CM differentiation (early knockdown) or in 3D engineered heart tissues as a disease model (late knockdown). hESC-CMs expressed the apelin signalling system at a similar level to the adult heart. Early apelin receptor knockdown decreased cardiomyocyte differentiation efficiency and prolonged voltage sensing, associated with asynchronous contraction. Late apelin receptor knockdown had detrimental consequences on 3D engineered heart tissue contractile properties, decreasing contractility and increasing stiffness. CONCLUSIONS: We have successfully knocked down the apelin receptor, using an inducible system, to demonstrate a key role in hESC-CM differentiation. Knockdown in 3D engineered heart tissues recapitulated the phenotype of apelin receptor down-regulation in a failing heart, providing a potential platform for modelling heart failure and testing novel therapeutic strategies.
Keywords
Apelin receptor, Cardiomyocyte, Cardiovascular disease, Stem cell
Sponsorship
British Heart Foundation (FS/17/61/33473)
TG/18/4/33770; FS/18/46/33663, RM/17/2/33380); Wellcome Trust (206194; 108413/A/15/D); Wellcome Trust Programme in Metabolic and Cardiovascular Disease (203814/Z/16/A) Wellcome Trust (203151/Z/16/Z), UKRI Medical Research Council (MC_PC_17230), Kusuma Trust, . Cambridge Biomedical Research Centre Biomedical Resources Grant (University of Cambridge, Cardiovascular Theme, RG64226).
Funder references
British Heart Foundation (TG/18/4/33770)
British Heart Foundation (FS/17/61/33473D)
Medical Research Council (MC_PC_17230)
Identifiers
External DOI: https://doi.org/10.1093/cvr/cvac065
This record's URL: https://www.repository.cam.ac.uk/handle/1810/339762
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