Spatiotemporal Cell Junction Assembly in Human IPSC-CM Models of Arrhythmogenic Cardiomyopathy

Overview

Journal Stem Cell Reports

Publisher Cell Press

Specialty Cell Biology

Date 2023 Aug 18

PMID 37595583

Authors

Sean L Kim

Michael A Trembley

Keel Yong Lee

Suji Choi

Luke A MacQueen

John F Zimmerman

Lousanne H C de Wit

Kevin Shani

Douglas E Henze

Daniel J Drennan

Shaila A Saifee

Li Jun Loh

Xujie Liu

Kevin Kit Parker

William T Pu

Affiliations

Soon will be listed here.

Abstract

Arrhythmogenic cardiomyopathy (ACM) is an inherited cardiac disorder that causes life-threatening arrhythmias and myocardial dysfunction. Pathogenic variants in Plakophilin-2 (PKP2), a desmosome component within specialized cardiac cell junctions, cause the majority of ACM cases. However, the molecular mechanisms by which PKP2 variants induce disease phenotypes remain unclear. Here we built bioengineered platforms using genetically modified human induced pluripotent stem cell-derived cardiomyocytes to model the early spatiotemporal process of cardiomyocyte junction assembly in vitro. Heterozygosity for truncating variant PKP2 reduced Wnt/β-catenin signaling, impaired myofibrillogenesis, delayed mechanical coupling, and reduced calcium wave velocity in engineered tissues. These abnormalities were ameliorated by SB216763, which activated Wnt/β-catenin signaling, improved cytoskeletal organization, restored cell junction integrity in cell pairs, and improved calcium wave velocity in engineered tissues. Together, these findings highlight the therapeutic potential of modulating Wnt/β-catenin signaling in a human model of ACM.

Citing Articles

Genetic and Molecular Underpinnings of Atrial Fibrillation.

Sweat M, Pu W NPJ Cardiovasc Health. 2025; 1.

PMID: 39867228 PMC: 11759492. DOI: 10.1038/s44325-024-00035-5.

Targeting Canonical Wnt-signaling Through GSK-3β in Arrhythmogenic Cardiomyopathy: Conservative or Progressive?.

Low B, Asimaki A J Cardiovasc Transl Res. 2024; 18(1):121-132.

PMID: 39392548 PMC: 11885336. DOI: 10.1007/s12265-024-10567-x.

hiPSC-derived cardiomyocytes as a model to study the role of small-conductance Ca-activated K (SK) ion channel variants associated with atrial fibrillation.

Babini H, Jimenez-Sabado V, Stogova E, Arslanova A, Butt M, Dababneh S Front Cell Dev Biol. 2024; 12:1298007.

PMID: 38304423 PMC: 10830749. DOI: 10.3389/fcell.2024.1298007.

References

Manring H, Dorn L, Ex-Willey A, Accornero F, Ackermann M . At the heart of inter- and intracellular signaling: the intercalated disc. Biophys Rev. 2018; 10(4):961-971. PMC: 6082301. DOI: 10.1007/s12551-018-0430-7. View

Chang H, Liu Q, Zimmerman J, Lee K, Jin Q, Peters M . Recreating the heart's helical structure-function relationship with focused rotary jet spinning. Science. 2022; 377(6602):180-185. PMC: 10077766. DOI: 10.1126/science.abl6395. View

Grosberg A, Kuo P, Guo C, Geisse N, Bray M, Adams W . Self-organization of muscle cell structure and function. PLoS Comput Biol. 2011; 7(2):e1001088. PMC: 3044763. DOI: 10.1371/journal.pcbi.1001088. View

Aratyn-Schaus Y, Pasqualini F, Yuan H, McCain M, Ye G, Sheehy S . Coupling primary and stem cell-derived cardiomyocytes in an in vitro model of cardiac cell therapy. J Cell Biol. 2016; 212(4):389-97. PMC: 4754718. DOI: 10.1083/jcb.201508026. View

Austin K, Trembley M, Chandler S, Sanders S, Saffitz J, Abrams D . Molecular mechanisms of arrhythmogenic cardiomyopathy. Nat Rev Cardiol. 2019; 16(9):519-537. PMC: 6871180. DOI: 10.1038/s41569-019-0200-7. View

Zhang K, Cloonan P, Sundaram S, Liu F, Das S, Ewoldt J . Plakophilin-2 truncating variants impair cardiac contractility by disrupting sarcomere stability and organization. Sci Adv. 2021; 7(42):eabh3995. PMC: 8519574. DOI: 10.1126/sciadv.abh3995. View

Bhonsale A, Groeneweg J, James C, Dooijes D, Tichnell C, Jongbloed J . Impact of genotype on clinical course in arrhythmogenic right ventricular dysplasia/cardiomyopathy-associated mutation carriers. Eur Heart J. 2015; 36(14):847-55. DOI: 10.1093/eurheartj/ehu509. View

Lian X, Zhang J, Azarin S, Zhu K, Hazeltine L, Bao X . Directed cardiomyocyte differentiation from human pluripotent stem cells by modulating Wnt/β-catenin signaling under fully defined conditions. Nat Protoc. 2012; 8(1):162-75. PMC: 3612968. DOI: 10.1038/nprot.2012.150. View

Saffitz J, Kleber A . Effects of mechanical forces and mediators of hypertrophy on remodeling of gap junctions in the heart. Circ Res. 2004; 94(5):585-91. DOI: 10.1161/01.RES.0000121575.34653.50. View

10.

Lee K, Park S, Matthews D, Kim S, Marquez C, Zimmerman J . An autonomously swimming biohybrid fish designed with human cardiac biophysics. Science. 2022; 375(6581):639-647. PMC: 8939435. DOI: 10.1126/science.abh0474. View

11.

Kobayashi T, Hino S, Oue N, Asahara T, Zollo M, Yasui W . Glycogen synthase kinase 3 and h-prune regulate cell migration by modulating focal adhesions. Mol Cell Biol. 2006; 26(3):898-911. PMC: 1347031. DOI: 10.1128/MCB.26.3.898-911.2006. View

12.

Sheehy S, Pasqualini F, Grosberg A, Park S, Aratyn-Schaus Y, Parker K . Quality metrics for stem cell-derived cardiac myocytes. Stem Cell Reports. 2014; 2(3):282-94. PMC: 3964283. DOI: 10.1016/j.stemcr.2014.01.015. View

13.

Chelko S, Asimaki A, Andersen P, Bedja D, Amat-Alarcon N, DeMazumder D . Central role for GSK3β in the pathogenesis of arrhythmogenic cardiomyopathy. JCI Insight. 2016; 1(5). PMC: 4861310. DOI: 10.1172/jci.insight.85923. View

14.

Syrris P, Ward D, Asimaki A, Sen-Chowdhry S, Ebrahim H, Evans A . Clinical expression of plakophilin-2 mutations in familial arrhythmogenic right ventricular cardiomyopathy. Circulation. 2006; 113(3):356-64. DOI: 10.1161/CIRCULATIONAHA.105.561654. View

15.

Thomas D, Choi S, Alamana C, Parker K, Wu J . Cellular and Engineered Organoids for Cardiovascular Models. Circ Res. 2022; 130(12):1780-1802. DOI: 10.1161/CIRCRESAHA.122.320305. View

16.

Kim C, Wong J, Wen J, Wang S, Wang C, Spiering S . Studying arrhythmogenic right ventricular dysplasia with patient-specific iPSCs. Nature. 2013; 494(7435):105-10. PMC: 3753229. DOI: 10.1038/nature11799. View

17.

Wang G, Yang L, Grishin D, Rios X, Ye L, Hu Y . Efficient, footprint-free human iPSC genome editing by consolidation of Cas9/CRISPR and piggyBac technologies. Nat Protoc. 2016; 12(1):88-103. PMC: 5352979. DOI: 10.1038/nprot.2016.152. View

18.

Sato P, Musa H, Coombs W, Guerrero-Serna G, Patino G, Taffet S . Loss of plakophilin-2 expression leads to decreased sodium current and slower conduction velocity in cultured cardiac myocytes. Circ Res. 2009; 105(6):523-6. PMC: 2742576. DOI: 10.1161/CIRCRESAHA.109.201418. View

19.

Vite A, Radice G . N-cadherin/catenin complex as a master regulator of intercalated disc function. Cell Commun Adhes. 2014; 21(3):169-79. PMC: 6054126. DOI: 10.3109/15419061.2014.908853. View

20.

Khudiakov A, Zaytseva A, Perepelina K, Smolina N, Pervunina T, Vasichkina E . Sodium current abnormalities and deregulation of Wnt/β-catenin signaling in iPSC-derived cardiomyocytes generated from patient with arrhythmogenic cardiomyopathy harboring compound genetic variants in plakophilin 2 gene. Biochim Biophys Acta Mol Basis Dis. 2020; 1866(11):165915. DOI: 10.1016/j.bbadis.2020.165915. View