TY - JOUR
T1 - PITX2 induction leads to impaired cardiomyocyte function in arrhythmogenic cardiomyopathy
AU - van Kampen, Sebastiaan J
AU - Han, Su Ji
AU - van Ham, Willem B
AU - Kyriakopoulou, Eirini
AU - Stouthart, Elizabeth W
AU - Goversen, Birgit
AU - Monshouwer-Kloots, Jantine
AU - Perini, Ilaria
AU - de Ruiter, Hesther
AU - van der Kraak, Petra
AU - Vink, Aryan
AU - van Laake, Linda W
AU - Groeneweg, Judith A
AU - de Boer, Teun P
AU - Tsui, Hoyee
AU - Boogerd, Cornelis J
AU - van Veen, Toon A B
AU - van Rooij, Eva
N1 - 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
PY - 2023/3/14
Y1 - 2023/3/14
N2 - 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 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.
AB - 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 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.
KW - arrhythmogenic cardiomyopathy
KW - cardiomyocyte
KW - desmoplakin
KW - desmosome
KW - function
KW - induced pluripotent stem cells
KW - PITX2
UR - http://www.scopus.com/inward/record.url?scp=85149904251&partnerID=8YFLogxK
U2 - 10.1016/j.stemcr.2023.01.015
DO - 10.1016/j.stemcr.2023.01.015
M3 - Article
C2 - 36868229
SN - 2213-6711
VL - 18
SP - 749
EP - 764
JO - Stem Cell Reports
JF - Stem Cell Reports
IS - 3
ER -