CRISPR correction of the PRKAG2 gene mutation in the patient's induced pluripotent stem cell-derived cardiomyocytes eliminates electrophysiological and structural abnormalities

Jehuda, Ronen Ben; Eisen, Binyamin; Shemer, Yuval; Mekies, Lucy N.; Szántai, Á.; Reiter, Irina; Cui, Huanhuan; Guan, Kaomei; Haron-Khun, Shiraz; Freimark, Dov; Sperling, Silke; Gherghiceanu, Mihaela; Arad, Michael; Binah, Ofer

doi:10.1016/j.hrthm.2017.09.024

Cited by 56 publications

(38 citation statements)

References 30 publications

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“…In those cases, targeted gene correction can reverse the pathological phenotype and normalize APD values. Gene editing in the context of reversing a pathological phenotype has been applied in multiple arrhythmogenic conditions including LQTS [113], SQTS [137], BrS [138], HCM [143], and Barth syndrome [150].…”

Section: Genome Editingmentioning

confidence: 99%

The Emergence of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes (hiPSC-CMs) as a Platform to Model Arrhythmogenic Diseases

Pourrier

Fedida

2020

IJMS

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There is a need for improved in vitro models of inherited cardiac diseases to better understand basic cellular and molecular mechanisms and advance drug development. Most of these diseases are associated with arrhythmias, as a result of mutations in ion channel or ion channel-modulatory proteins. Thus far, the electrophysiological phenotype of these mutations has been typically studied using transgenic animal models and heterologous expression systems. Although they have played a major role in advancing the understanding of the pathophysiology of arrhythmogenesis, more physiological and predictive preclinical models are necessary to optimize the treatment strategy for individual patients. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have generated much interest as an alternative tool to model arrhythmogenic diseases. They provide a unique opportunity to recapitulate the native-like environment required for mutated proteins to reproduce the human cellular disease phenotype. However, it is also important to recognize the limitations of this technology, specifically their fetal electrophysiological phenotype, which differentiates them from adult human myocytes. In this review, we provide an overview of the major inherited arrhythmogenic cardiac diseases modeled using hiPSC-CMs and for which the cellular disease phenotype has been somewhat characterized.

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Section: Genome Editingmentioning

confidence: 99%

The Emergence of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes (hiPSC-CMs) as a Platform to Model Arrhythmogenic Diseases

Pourrier

Fedida

2020

IJMS

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“…We deployed cellular micropatterning devices on hydrogel substrates to measure contractile properties of hiPSC-CMs using traction force microscopy (as described in Ribeiro, et al [28]). Previous studies of hiPSC-CMs and cardiac tissues derived from patients with HCM have shown contractile dysfunction, including contractile arrhythmias and hypercontractility [20,32,33], as well as greater developed force and shorter twitch duration [34], in patient-derived cardiomyocytes as compared to wild-type controls. In our study, we found that MYH7-R403Q hiPSC-CMs had significantly increased maximal contraction and relaxation velocities and powers, as well as significantly increased maximal force when compared to wild-type hiPSC-CMs.…”

Section: Discussionmentioning

confidence: 96%

Dissociation of disease phenotype and allele silencing in hypertrophic cardiomyopathy

Dainis

Zaleta-Rivera

Ribeiro

et al. 2019

Preprint

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Allele-specific RNA silencing has been shown to be an effective therapeutic treatment in a number of diseases, including neurodegenerative disorders. Studies of allele-specific silencing in hypertrophic cardiomyopathy to date have focused on mouse models of disease. Here, we investigate two methods of allele-specific silencing, short hairpin RNA (shRNA) and antisense oligonucleotide (ASO) silencing, using a human induced pluripotent stem cell-derived cardiomyocyte (hiPSC-CM) model of disease. We used cellular micropatterning devices with traction force microscopy and automated video analysis to examine each strategy's effects on contractile defects underlying disease. We find that shRNA silencing ameliorates contractile phenotypes of disease, reducing disease-associated increases in cardiomyocyte velocity, force, and power. We find that ASO silencing, while better able to target and knockdown a specific disease-associated allele, showed more modest improvements in contractile phenotypes. We find a dissociation between allelic-specificity and functional improvements between the two tested therapeutic strategies, suggesting a more complex method of allelic control underlying HCM-associated transcripts. Author summaryAllele-specific silencing, whereby a therapeutic molecule is used to lower the expression of just one of the two copies or alleles of a gene, may be a potential therapeutic strategy in diseases caused by a single mutation. In this paper, we examine two such strategies in hypertrophic cardiomyopathy, a disease characterized by an overgrowth of the left-ventricular heart muscle as well as contractile dysfunction. We used a human cell model of disease, creating induced pluripotent stem cell derived cardiomyocytes from a patient with HCM caused by a single base pair change in just one allele of the gene MYH7. We used two strategies to silence the disease-associated copy of MYH7, both focused on reducing RNA expression from the mutated allele, as well as state-of-the-art biophysical techniques for measuring contractility. We found that one silencing strategy, which reduced expression of both the disease-associated and the healthy alleles of MYH7, showed great improvements in contractility between treated and untreated cells. Our

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“…We annotated a significant promoter switch in PRKAG2, a gene linked to hypertrophic cardiomyopathy and critical to heart metabolism. [36][37][38] Three major PRKAG2 promoters were identified, encoding three different isoforms-g2a, g2-3b, and g2b ( Figure 5D).…”

Section: Correlation Of Cage-sequencing and Rna-sequencingmentioning

confidence: 99%

Enhancer and promoter usage in the normal and failed human heart

Am¹,

Dellefave‐Castillo²,

Pgt³

et al. 2020

Preprint

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Word count: 5331 (excluding references and legends)Short title: Promoter and enhancer shifts in heart failure ABSTRACTThe failed heart is characterized by re-expression of a fetal gene program, which contributes to adaptation and maladaptation in heart failure. To define genomewide enhancer and promoter use in heart failure, Cap Analysis of Gene Expression (CAGE-seq) was applied to healthy and failed human left ventricles to define short RNAs associated with both promoters and enhancers. Integration of CAGE-seq data with RNA sequencing identified a combined ~17,000 promoters and ~1,500 enhancers active in healthy and failed human left ventricles. Comparing promoter usage between healthy and failed hearts highlighted promoter shifts which altered amino-terminal protein sequences. Comparing enhancer usage between healthy and failed hearts revealed a majority of differentially utilized heart failure enhancers were intronic and primarily localized within the first intron, identifying this position as a common feature associated with tissue-specific gene expression changes in the heart. This dataset defines the dynamic genomic regulatory landscape underlying heart failure and serves as an important resource for understanding genetic contributions to cardiac dysfunction. Wang from the Northwestern University sequencing core and Yujiro Takegami from DNAFORM for their excellent technical support. AG conducted the analysis and drafted the mansuscript. LDC secured patient consent and genotype information. PP assisted with genotyping. JAW provided access to control samples. DYB and MJP provided helpful advice and commentary and assisted with interpretation. MAN and EMM assisted with analysis, writing and editing the manuscript.

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CRISPR correction of the PRKAG2 gene mutation in the patient's induced pluripotent stem cell-derived cardiomyocytes eliminates electrophysiological and structural abnormalities

Cited by 56 publications

References 30 publications

The Emergence of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes (hiPSC-CMs) as a Platform to Model Arrhythmogenic Diseases

The Emergence of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes (hiPSC-CMs) as a Platform to Model Arrhythmogenic Diseases

Dissociation of disease phenotype and allele silencing in hypertrophic cardiomyopathy

Enhancer and promoter usage in the normal and failed human heart

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