Altered Electrical, Biomolecular, and Immunologic Phenotypes in a Novel Patient-Derived Stem Cell Model of Desmoglein-2 Mutant ARVC

Journal of Clinical Medicine

Suraj Kannan¹  Adriana Blazeski¹  Leslie Tung¹  Justin Morrissette-McAlmon¹  Robert N. Hawthorne¹  Justin Lowenthal¹  Kenneth R. Boheler¹  Roald Teuben¹  Cynthia A. James²  Gordon Tomaselli²  Deborah DiSilvestre²  Stephen P. Chelko²  Jeffrey E. Saffitz³ 

[1] Department of Biomedical Engineering, School of Medicine, Johns Hopkins University, Baltimore, MD 21205, USA;Department of Medicine, School of Medicine, Johns Hopkins University, Baltimore, MD 21205, USA;Department of Pathology, Beth Israel Deaconess Medical Center, Boston, MA 02215, USA;

关键词: arrhythmogenic cardiomyopathy;    arrhythmogenic right ventricular cardiomyopathy;    ARVC;    desmoglein-2;    NFκB signaling;    patient-derived stem cells;   

DOI  :  10.3390/jcm10143061

来源: DOAJ

【 摘 要 】

Arrhythmogenic right ventricular cardiomyopathy (ARVC) is a progressive heart condition which causes fibro-fatty myocardial scarring, ventricular arrhythmias, and sudden cardiac death. Most cases of ARVC can be linked to pathogenic mutations in the cardiac desmosome, but the pathophysiology is not well understood, particularly in early phases when arrhythmias can develop prior to structural changes. Here, we created a novel human induced pluripotent stem cell-derived cardiomyocyte (hiPSC-CM) model of ARVC from a patient with a c.2358delA variant in desmoglein-2 (DSG2). These DSG2-mutant (DSG2^Mut) hiPSC-CMs were compared against two wildtype hiPSC-CM lines via immunostaining, RT-qPCR, Western blot, RNA-Seq, cytokine expression and optical mapping. Mutant cells expressed reduced DSG2 mRNA and had altered localization of desmoglein-2 protein alongside thinner, more disorganized myofibrils. No major changes in other desmosomal proteins were noted. There was increased pro-inflammatory cytokine expression that may be linked to canonical and non-canonical NFκB signaling. Action potentials in DSG2^Mut CMs were shorter with increased upstroke heterogeneity, while time-to-peak calcium and calcium decay rate were reduced. These were accompanied by changes in ion channel and calcium handling gene expression. Lastly, suppressing DSG2 in control lines via siRNA allowed partial recapitulation of electrical anomalies noted in DSG2^Mut cells. In conclusion, the aberrant cytoskeletal organization, cytokine expression, and electrophysiology found DSG2^Mut hiPSC-CMs could underlie early mechanisms of disease manifestation in ARVC patients.

【 授权许可】

Unknown