Stem Cell–Derived Cardiomyocytes and Beta-Adrenergic Receptor Blockade in Duchenne Muscular Dystrophy Cardiomyopathy
Authors: Satyabrata Das
Pruthvi Shah
Jianyi Zhang
James M Ervasti
Daniel J Garry
Forum Kamdar
André Klaassen Kamdar
Mary G Garry
Tatyana A Meyers
DeWayne Townsend
Joseph C Wu
Timothy J Kamp
Wuming Gong
Journal: Journal of the American College of Cardiology (2020)
Discipline: CLINICAL MEDICINE
Keywords: Duchenne disease modeling; heart failure; human inducible pluripotent stem cells; muscular dystrophy cardiomyopathy; β-blockers.
Affiliations: Lillehei Heart Institute, Cardiovascular Division, University of Minnesota, Minneapolis, Minnesota; Paul and Sheila Muscular Dystrophy Center, University of Minnesota, Minneapolis, Minnesota., Lillehei Heart Institute, Cardiovascular Division, University of Minnesota, Minneapolis, Minnesota., Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, Minnesota., Paul and Sheila Muscular Dystrophy Center, University of Minnesota, Minneapolis, Minnesota; Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, Minnesota., Lillehei Heart Institute, Cardiovascular Division, University of Minnesota, Minneapolis, Minnesota; Paul and Sheila Muscular Dystrophy Center, University of Minnesota, Minneapolis, Minnesota; Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, Minnesota., Cardiovascular Medicine, University of Wisconsin-Madison, Madison, Wisconsin., Stanford Cardiovascular Institute, Stanford University, Palo Alto, California., Department of Biomedical Engineering, University of Alabama at Birmingham, Birmingham, Alabama., Lillehei Heart Institute, Cardiovascular Division, University of Minnesota, Minneapolis, Minnesota; Paul and Sheila Muscular Dystrophy Center, University of Minnesota, Minneapolis, Minnesota; Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, Minnesota; Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota. Electronic address: .

Abstract:  Background: Although cardiomyopathy has emerged as a leading cause of death in Duchenne muscular dystrophy (DMD), limited studies and therapies have emerged for dystrophic heart failure. Objectives: The purpose of this study was to model DMD cardiomyopathy using DMD patient-specific human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes and to identify physiological changes and future drug therapies. Methods: To explore and define therapies for DMD cardiomyopathy, the authors used DMD patient-specific hiPSC-derived cardiomyocytes to examine the physiological response to adrenergic agonists and β-blocker treatment. The authors further examined these agents in vivo using wild-type and mdx mouse models. Results: At baseline and following adrenergic stimulation, DMD hiPSC-derived cardiomyocytes had a significant increase in arrhythmic calcium traces compared to isogenic controls. Furthermore, these arrhythmias were significantly decreased with propranolol treatment. Using telemetry monitoring, the authors observed that mdx mice, which lack dystrophin, had an arrhythmic death when stimulated with isoproterenol; the lethal arrhythmias were rescued, in part, by propranolol pre-treatment. Using single-cell and bulk RNA sequencing (RNA-seq), the authors compared DMD and control hiPSC-derived cardiomyocytes, mdx mice, and control mice (in the presence or absence of propranolol and isoproterenol) and defined pathways that were perturbed under baseline conditions and pathways that were normalized after propranolol treatment in the mdx model. The authors also undertook transcriptome analysis of human DMD left ventricle samples and found that DMD hiPSC-derived cardiomyocytes have dysregulated pathways similar to the human DMD heart. The authors further determined that relatively few patients with DMD see a cardiovascular specialist or receive β-blocker therapy. Conclusions: The results highlight mechanisms and therapeutic interventions from human to animal and back to human in the dystrophic heart. These results may serve as a prelude for an adequately powered clinical study that examines the impact of β-blocker therapy in patients with dystrophinopathies. 

Retracted: No

PubPeer comments:
 There is a general lack of information regarding the origins of their hiPSC lines, most notably their patient derived DMD lines. The authors do not indicate obtaining the cells from outside their lab(s) and discuss cite fibroblast reprogramming protocols, which gives the impression that they personally developed the cell lines DMD1 and DMD2 from patients. In addition, discussing MLPA methods to confirm identity of specific DMD mutations reinforces this idea. However, Online Figure 2 has additional identifiers for these cell lines: SC604 and 5169. These identifiers appear to match those of DMD hiPSC lines available from System Biosciences and the Coriell Institute (details below). In addition, the mutations described in the manuscript (deletion of exons 3-7 and deletion of exons 4-43) match the description of the System Biosciences and Coriell Institute cell lines. The authors also do not discuss the origins of the control cells in these studies, or those used in for the gene-edited DMD3 and DMD4. SC604A B available from System Biosciences https: bioscience.co.uk userfiles pdf disease_ips_cells_web.pdf https: web.expasy.org cellosaurus CVCL_IT72 A cell line identified as GM05169 is available through the Coriell Institute https: www.coriell.org 0 Sections Search Sample_Detail.aspx?Ref=GM05169 product=CC , 
 #1 I completely agree that this paper does not describe well enough the origin of their iPSC cell lines. I however want to point out that Coriell GM05169 is not an iPSC line but a skin fibroblast cell line. It is therefore possible that the authors have derived an iPSC from these cells. Other groups have already done so from the same cell line. See: https: web.expasy.org cgi-bin cellosaurus search?input=GM05169 But this comment is not valid for SC604 and as you stated they do not give any details on the protocols followed to establish their iPSCs nor does it provide information on the patients age and sex. , 
 I noticed that the supplementary material on the JACC and Elsevier websites has been recently updated now to address the origins of the cell lines used in the study, now including a new section of text:  hiPSC lines To address the need for an in vitro model of DMD cardiomyopathy we established hiPSC cardiomyocytes from patients with clinically diagnosed Duchenne Muscular Dystrophy. DMD1 was purchased from Systems Biosciences, Palo Alto, CA and DMD2 was purchased from Coriell Institute, Camden, NJ. The control lines were obtained from iPierian. iDMD3 and iDMD4 were gene engineered lines based on control 1. *** the meta data for supplement Word document indicates that it was last edited on 8-30-2021, whereas the original document that did not include this passage was last edited 1-30-2020. However, this significant addition has surprisingly not been accompanied by an official Correction Notice from the journal. Also, while the newly included supplemental information somewhat resolves issues related to the JACC manuscript itself (still too vague and somewhat inconsistent with text in the main body of the manuscript), it also seems to more directly contradict several conference abstracts previously published by several of the authors. https: www.jacc.org doi 10.1016 S0735-1097%2815%2962149-0 https: www.ahajournals.org doi 10.1161 circ.128.suppl_22.A17079 https: www.ahajournals.org doi 10.1161 circ.132.suppl_3.18056 All these abstracts are a part of the same study detailed in the JACC manuscript and include some phrasing along the lines of  Dermal fibroblasts were obtained from patients with DMD cardiomyopathy (dystrophin deletion of exon 4-43) and normal healthy controls. The fibroblasts were reprogrammed to hiPSC using retroviral vectors containing the human transcription factors, OCT4, SOX2, KLF4 and C-MYC. Yet, the new supplemental material explicitly states that the control and disease cell lines were purchased obtained from external sources, likely already reprogrammed into hiPSC. ,