Electrophysiologic Characterization of Calcium Handling in Human Induced Pluripotent Stem Cell-Derived Atrial Cardiomyocytes

Argenziano, Mariana; Lambers, Erin; Luan, Hong; Sridhar, Arvind; Zhang, Meihong; Chalazan, Brandon; Menon, Ambili; Savio‐Galimberti, Eleonora; Wu, Joseph C.; Rehman, Jalees; Darbar, Dawood

doi:10.1016/j.stemcr.2018.04.005

Cited by 51 publications

(65 citation statements)

References 32 publications

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“…Overall, the APs of our atrial-like hiPSC-CMs were substantially shorter and had a lower AP plateau than those of our ventricular-like hiPSC-CMs, in qualitative agreement with previous studies on atrial- and ventricular-like hiPSC-CMs ( Marczenke et al, 2017 ; Verkerk et al, 2017 ; Argenziano et al, 2018 ; Cyganek et al, 2018 ; Lemme et al, 2018 ; Veerman et al, 2019 ). There are some quantitative differences in AP parameters with previous studies, but these are likely due to differences in cell lines, differences in differentiation protocols, absence or presence of I K 1 injection, and a different definition of AP plateau amplitude.…”

Section: Discussionsupporting

confidence: 92%

Ultrarapid Delayed Rectifier K+ Channelopathies in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

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Devalla

Bosch

et al. 2020

Front. Cell Dev. Biol.

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Atrial fibrillation (AF) is the most common cardiac arrhythmia. About 5–15% of AF patients have a mutation in a cardiac gene, including mutations in KCNA5 , encoding the K v 1.5 α-subunit of the ion channel carrying the atrial-specific ultrarapid delayed rectifier K + current (I Kur ). Both loss-of-function and gain-of-function AF-related mutations in KCNA5 are known, but their effects on action potentials (APs) of human cardiomyocytes have been poorly studied. Here, we assessed the effects of wild-type and mutant I Kur on APs of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). We found that atrial-like hiPSC-CMs, generated by a retinoic acid-based differentiation protocol, have APs with faster repolarization compared to ventricular-like hiPSC-CMs, resulting in shorter APs with a lower AP plateau. Native I Kur , measured as current sensitive to 50 μM 4-aminopyridine, was 1.88 ± 0.49 (mean ± SEM, n = 17) and 0.26 ± 0.26 pA/pF ( n = 17) in atrial- and ventricular-like hiPSC-CMs, respectively. In both atrial- and ventricular-like hiPSC-CMs, I Kur blockade had minimal effects on AP parameters. Next, we used dynamic clamp to inject various amounts of a virtual I Kur , with characteristics as in freshly isolated human atrial myocytes, into 11 atrial-like and 10 ventricular-like hiPSC-CMs, in which native I Kur was blocked. Injection of I Kur with 100% density shortened the APs, with its effect being strongest on the AP duration at 20% repolarization (APD 20 ) of atrial-like hiPSC-CMs. At I Kur densities < 100% (compared to 100%), simulating loss-of-function mutations, significant AP prolongation and raise of plateau were observed. At I Kur densities > 100%, simulating gain-of-function mutations, APD 20 was decreased in both atrial- and ventricular-like hiPSC-CMs, but only upon a strong increase in I Kur . In ventricular-like hiPSC-CMs, lowering of the plateau resulted in AP shortening. We conclude that a decrease in I Kur , mimicking loss-of-function mutations, has a stronger effect on the AP of hiPSC-CMs than an increase, mimicking gain-of-function mutations, whereas in ventricular-like hiPSC-CMs such increase results in AP shortening, causing their AP morphology to become more atrial-like. Effects of native I Kur modulation on atrial-like hiPSC-CMs are less pronounced than effects of virtual I Kur injection because I Kur density of atrial-like hiPSC-CMs is substantially smaller than that of freshly isolated human atrial myocytes.

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Section: Discussionsupporting

confidence: 92%

Ultrarapid Delayed Rectifier K+ Channelopathies in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Hilderink

Devalla

Bosch

et al. 2020

Front. Cell Dev. Biol.

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“…Potentially, this could have an effect on the plateau phase of the AP. It has also been reported that atrial-like hiPSC-CMs have lower expression of CACNA1C, as well as reduced peak I Ca,L amplitude and current density [49]. However, the precise functional roles of the antisense and intronic transcripts we identified remain unclear.…”

Section: Discussionmentioning

confidence: 81%

Single-Cell RNA-Sequencing and Optical Electrophysiology of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes Reveal Discordance Between Cardiac Subtype-Associated Gene Expression Patterns and Electrophysiological Phenotypes

Biendarra-Tiegs

et al. 2019

Stem Cells and Development

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The ability to accurately phenotype cells differentiated from human induced pluripotent stem cells (hiPSCs) is essential for their application in modeling developmental and disease processes, yet also poses a particular challenge without the context of anatomical location. Our specific objective was to determine if single-cell gene expression was sufficient to predict the electrophysiology of iPSC-derived cardiac lineages, to evaluate the concordance between molecular and functional surrogate markers. To this end, we used the genetically encoded voltage indicator ArcLight to profile hundreds of hiPSC-derived cardiomyocytes (hiPSC-CMs), thus identifying patterns of electrophysiological maturation and increased prevalence of cells with atrial-like action potentials (APs) between days 11 and 42 of differentiation. To profile expression patterns of cardiomyocyte subtypeassociated genes, single-cell RNA-seq was performed at days 12 and 40 after the populations were fully characterized with the high-throughput ArcLight platform. Although we could detect global gene expression changes supporting progressive differentiation, individual cellular expression patterns alone were not able to delineate the individual cardiomyocytes into atrial, ventricular, or nodal subtypes as functionally documented by electrophysiology measurements. Furthermore, our efforts to understand the distinct electrophysiological properties associated with day 12 versus day 40 hiPSC-CMs revealed that ion channel regulators SLMAP, FGF12, and FHL1 were the most significantly increased genes at day 40, categorized by electrophysiologyrelated gene functions. Notably, FHL1 knockdown during differentiation was sufficient to significantly modulate APs toward ventricular-like electrophysiology. Thus, our results establish the inability of subtypeassociated gene expression patterns to specifically categorize hiPSC-derived cells according to their functional electrophysiology, and yet, altered FHL1 expression is able to redirect electrophysiological maturation of these developing cells. Therefore, noncanonical gene expression patterns of cardiac maturation may be sufficient to direct functional maturation of cardiomyocytes, with canonical gene expression patterns being insufficient to temporally define cardiac subtypes of in vitro differentiation.

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“…Coupling between CF and cardiomyocytes can either promote or suppress cardiac alternans, depending on two competing effects on repolarization and Ca 2+ cycling [ 69 ]. Further, cardiomyocytes with a relatively immature set of calcium-handling proteins, as is likely found in hiPSC-CMs [ 70 ], may be more prone to alternans due to an inability to effectively regulate calcium current ( I Ca ), which is a major driver of alternans [ 71 ]. In our data set, the pacing frequency at which alternans onset occurs increases with dose-dependent hCF inclusion ( Figure 5(c) ), likely due to the higher maximum capture rates of higher percentage hCF tissues.…”

Section: Discussionmentioning

confidence: 99%

Human Cardiac Fibroblast Number and Activation State Modulate Electromechanical Function of hiPSC-Cardiomyocytes in Engineered Myocardium

Rupert

Kim

Choi

et al. 2020

Stem Cells International

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Cardiac tissue engineering using hiPSC-derived cardiomyocytes is a promising avenue for cardiovascular regeneration, pharmaceutical drug development, cardiotoxicity evaluation, and disease modeling. Limitations to these applications still exist due in part to the need for more robust structural support, organization, and electromechanical function of engineered cardiac tissues. It is well accepted that heterotypic cellular interactions impact the phenotype of cardiomyocytes. The current study evaluates the functional effects of coculturing adult human cardiac fibroblasts (hCFs) in 3D engineered tissues on excitation and contraction with the goal of recapitulating healthy, nonarrhythmogenic myocardium in vitro. A small population (5% of total cell number) of hCFs in tissues improves tissue formation, material properties, and contractile function. However, two perturbations to the hCF population create disease-like phenotypes in engineered cardiac tissues. First, increasing the percentage of hCFs to 15% resulted in tissues with increased ectopic activity and spontaneous excitation rate. Second, hCFs undergo myofibroblast activation in traditional two-dimensional culture, and this altered phenotype ablated the functional benefits of hCFs when incorporated into engineered cardiac tissues. Taken together, the results of this study demonstrate that human cardiac fibroblast number and activation state modulate electromechanical function of hiPSC-cardiomyocytes and that a low percentage of quiescent hCFs are a valuable cell source to advance a healthy electromechanical response of engineered cardiac tissue for regenerative medicine applications.

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Electrophysiologic Characterization of Calcium Handling in Human Induced Pluripotent Stem Cell-Derived Atrial Cardiomyocytes

Cited by 51 publications

References 32 publications

Ultrarapid Delayed Rectifier K+ Channelopathies in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Ultrarapid Delayed Rectifier K+ Channelopathies in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Single-Cell RNA-Sequencing and Optical Electrophysiology of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes Reveal Discordance Between Cardiac Subtype-Associated Gene Expression Patterns and Electrophysiological Phenotypes

Human Cardiac Fibroblast Number and Activation State Modulate Electromechanical Function of hiPSC-Cardiomyocytes in Engineered Myocardium

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