Article

Patient-Specific Induced Pluripotent Stem Cells as a Model for Familial Dilated Cardiomyopathy

Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Science translational medicine (Impact Factor: 15.84). 04/2012; 4(130):130ra47. DOI: 10.1126/scitranslmed.3003552
Source: PubMed

ABSTRACT

Characterized by ventricular dilatation, systolic dysfunction, and progressive heart failure, dilated cardiomyopathy (DCM) is the most common form of cardiomyopathy in patients. DCM is the most common diagnosis leading to heart transplantation and places a significant burden on healthcare worldwide. The advent of induced pluripotent stem cells (iPSCs) offers an exceptional opportunity for creating disease-specific cellular models, investigating underlying mechanisms, and optimizing therapy. Here, we generated cardiomyocytes from iPSCs derived from patients in a DCM family carrying a point mutation (R173W) in the gene encoding sarcomeric protein cardiac troponin T. Compared to control healthy individuals in the same family cohort, cardiomyocytes derived from iPSCs from DCM patients exhibited altered regulation of calcium ion (Ca(2+)), decreased contractility, and abnormal distribution of sarcomeric α-actinin. When stimulated with a β-adrenergic agonist, DCM iPSC-derived cardiomyocytes showed characteristics of cellular stress such as reduced beating rates, compromised contraction, and a greater number of cells with abnormal sarcomeric α-actinin distribution. Treatment with β-adrenergic blockers or overexpression of sarcoplasmic reticulum Ca(2+) adenosine triphosphatase (Serca2a) improved the function of iPSC-derived cardiomyocytes from DCM patients. Thus, iPSC-derived cardiomyocytes from DCM patients recapitulate to some extent the morphological and functional phenotypes of DCM and may serve as a useful platform for exploring disease mechanisms and for drug screening.

Download full-text

Full-text

Available from: Leng Han
  • Source
    • "In the cardiac field, hiPSC lines were established from healthy individuals (Zhang et al., 2009; Zwi et al., 2009) and from patients inflicted with acquired (heart failure) (Zwi-Dantsis et al., 2013) and inherited cardiac disorders. Among the latter, patient-specific hiPSC-derived cardiomyocytes (hiPSC-CMs) models of different inherited arrhythmogenic syndromes (Bellin et al., 2013; Caspi et al., 2013; Itzhaki et al., 2011a, 2012; Jung et al., 2012; Moretti et al., 2010) and diverse cardiomyopathies (Lan et al., 2013; Sun et al., 2012) were established. The patient/disease-specific hiPSC-CMs were shown to recapitulate the disease phenotypes in culture, to provide mechanistic insights into disease processes, and to evaluate existing and novel therapies. "
    [Show abstract] [Hide abstract]
    ABSTRACT: The advent of the human-induced pluripotent stem cell (hiPSC) technology has transformed biomedical research, providing new tools for human disease modeling, drug development, and regenerative medicine. To fulfill its unique potential in the cardiovascular field, efficient methods should be developed for high-resolution, large-scale, long-term, and serial functional cellular phenotyping of hiPSC-derived cardiomyocytes (hiPSC-CMs). To achieve this goal, we combined the hiPSC technology with genetically encoded voltage (ArcLight) and calcium (GCaMP5G) fluorescent indicators. Expression of ArcLight and GCaMP5G in hiPSC-CMs permitted to reliably follow changes in transmembrane potential and intracellular calcium levels, respectively. This allowed monitoring short- and long-term changes in action-potential and calcium-handling properties and the development of arrhythmias in response to several pharmaceutical agents and in hiPSC-CMs derived from patients with different inherited arrhythmogenic syndromes. Combining genetically encoded fluorescent reporters with hiPSC-CMs may bring a unique value to the study of inherited disorders, developmental biology, and drug development and testing.
    Full-text · Article · Sep 2015 · Stem Cell Reports
  • Source
    • "DCM iPSC-CMs Exhibit Impaired Response to b-Adrenergic Stimulation As abnormal b-adrenergic regulation in cardiac diseases has been well documented (Lohse et al., 2003; Post et al., 1999), we next examined whether DCM iPSC-CMs can also recapitulate this dysfunction. To address this question, we differentiated iPSC-CMs from both familial Ctrl and DCM (TNNT2 R173W) groups as described (Sun et al., 2012). Flow cytometry assays indicated that the efficiency of iPSC-CM differentiation in n = 3 Ctrl and n = 3 DCM patients was >90% (Figures S4A–S4C). "
    [Show abstract] [Hide abstract]
    ABSTRACT: β-adrenergic signaling pathways mediate key aspects of cardiac function. Its dysregulation is associated with a range of cardiac diseases, including dilated cardiomyopathy (DCM). Previously, we established an iPSC model of familial DCM from patients with a mutation in TNNT2, a sarcomeric protein. Here, we found that the β-adrenergic agonist isoproterenol induced mature β-adrenergic signaling in iPSC-derived cardiomyocytes (iPSC-CMs) but that this pathway was blunted in DCM iPSC-CMs. Although expression levels of several β-adrenergic signaling components were unaltered between control and DCM iPSC-CMs, we found that phosphodiesterases (PDEs) 2A and PDE3A were upregulated in DCM iPSC-CMs and that PDE2A was also upregulated in DCM patient tissue. We further discovered increased nuclear localization of mutant TNNT2 and epigenetic modifications of PDE genes in both DCM iPSC-CMs and patient tissue. Notably, pharmacologic inhibition of PDE2A and PDE3A restored cAMP levels and ameliorated the impaired β-adrenergic signaling of DCM iPSC-CMs, suggesting therapeutic potential. Copyright © 2015 Elsevier Inc. All rights reserved.
    Full-text · Article · Jun 2015 · Cell stem cell
  • Source
    • "Embryoid bodies (EBs) were formed from the control and DMD iPSCs (Fig. 2A) to induce cardiomyocyte differentiation using our previously established method (Lin et al., 2012) (supplementary material Fig. S2a,b). This cardiac differentiation method was developed from our previous cardiovascular differentiation protocol in human embryonic stem cells (ESCs) (Yang et al., 2008) and has been utilized for modeling Leopard-syndrome-associated hypertrophic cardiomyopathy (Carvajal-Vergara et al., 2010), dilated cardiomyopathy (Sun et al., 2012) and familiar hypertrophic cardiomyopathy (Lan et al., 2013) with patient-specific iPSCs. At day 22 of differentiation, iPSC-derived EBs exhibited spontaneous contractions (supplementary material Movies 1-3). "

    Full-text · Dataset · May 2015
Show more