PITX2 induction leads to impaired cardiomyocyte function in arrhythmogenic cardiomyopathy
van Kampen, Sebastiaan J; Han, Su Ji; van Ham, Willem B; Kyriakopoulou, Eirini; Stouthart, Elizabeth W; Goversen, Birgit; Monshouwer-Kloots, Jantine; Perini, Ilaria; de Ruiter, Hesther; van der Kraak, Petra; Vink, Aryan; van Laake, Linda W; Groeneweg, Judith A; de Boer, Teun P; Tsui, Hoyee; Boogerd, Cornelis J; van Veen, Toon A B; van Rooij, Eva
(2023) Stem Cell Reports, volume 18, issue 3, pp. 749 - 764
(Article)
Abstract
Arrhythmogenic cardiomyopathy (ACM) is an inherited progressive disease characterized by electrophysiological and structural remodeling of the ventricles. However, the disease-causing molecular pathways, as a consequence of desmosomal mutations, are poorly understood. Here, we identified a novel missense mutation within desmoplakin in a patient clinically diagnosed with ACM. Using CRISPR-Cas9, we
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corrected this mutation in patient-derived human induced pluripotent stem cells (hiPSCs) and generated an independent knockin hiPSC line carrying the same mutation. Mutant cardiomyocytes displayed a decline in connexin 43, NaV1.5, and desmosomal proteins, which was accompanied by a prolonged action potential duration. Interestingly, paired-like homeodomain 2 (PITX2), a transcription factor that acts a repressor of connexin 43, NaV1.5, and desmoplakin, was induced in mutant cardiomyocytes. We validated these results in control cardiomyocytes in which PITX2 was either depleted or overexpressed. Importantly, knockdown of PITX2 in patient-derived cardiomyocytes is sufficient to restore the levels of desmoplakin, connexin 43, and NaV1.5.
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Keywords: arrhythmogenic cardiomyopathy, cardiomyocyte, desmoplakin, desmosome, function, induced pluripotent stem cells, PITX2, Genetics, Biochemistry, Cell Biology, Developmental Biology, Journal Article
ISSN: 2213-6711
Publisher: Cell Press
Note: Funding Information: We thank V. Sigurdsson and the department of dermatology, UMC Utrecht, for help with skin biopsies. We thank Farhad A. Moqadam and Ana Rita Leitoguinho for the design of the gRNAs. We also thank Sjoerd J. Klaasen and Geert J.P.L. Kops for their help with the karyo sequencing procedure. This work was supported by funds from the Dutch Cardiovascular Alliance with the support of the Dutch Heart Foundation, DCVA2017-18 ARENA-PRIME (E.v.R.) and CVON2018-30 PREDICT2 (W.B.v.H. and T.A.B.v.V.); the Vici grant from the Dutch Research Council (NWO), project 09150181910020 (E.v.R.); and by the Fondation Leducq Transatlantic Network of Excellence, 17CVD02 (E.v.R.). The authors declare no competing interests. Funding Information: We thank V. Sigurdsson and the department of dermatology, UMC Utrecht, for help with skin biopsies. We thank Farhad A. Moqadam and Ana Rita Leitoguinho for the design of the gRNAs. We also thank Sjoerd J. Klaasen and Geert J.P.L. Kops for their help with the karyo sequencing procedure. This work was supported by funds from the Dutch Cardiovascular Alliance with the support of the Dutch Heart Foundation , DCVA2017-18 ARENA-PRIME (E.v.R.) and CVON2018-30 PREDICT2 (W.B.v.H. and T.A.B.v.V.); the Vici grant from the Dutch Research Council (NWO), project 09150181910020 (E.v.R.); and by the Fondation Leducq Transatlantic Network of Excellence, 17CVD02 (E.v.R.). Publisher Copyright: © 2023 The Authors
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