Drug Screening Using a Library of Human Induced Pluripotent Stem Cell–Derived Cardiomyocytes Reveals Disease-Specific Patterns of Cardiotoxicity

Liang, Ping; Lan, Feng; Lee, Andrew S.; Gong, Tingyu; Sánchez-Freire, Verónica; Wang, Yongming; Diecke, Sebastian; Sallam, Karim; Knowles, Joshua; Wang, Paul J.; Nguyen, Patricia K.; Bers, Donald M.; Robbins, Robert C.; Wu, Joseph C.

doi:10.1161/circulationaha.113.001883

Cited by 494 publications

(433 citation statements)

References 46 publications

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“…Despite their utility in human arrhythmia models, iPSC-CMs differ from adult tissue-derived cardiomyocytes (29)(30)(31)(32)(33). APs recorded from iPSC-CMs display depolarized diastolic potentials, myogenic AP firing, and a reduced maximum rise rate of the AP upstroke (dV/dt MAX ) compared with APs from adult ventricular tissue (Fig.…”

Section: Discussionmentioning

confidence: 99%

hERG 1b is critical for human cardiac repolarization

Jones¹,

Liu²,

Vaidyanathan

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

108

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The human ether-à-go-go-related gene (hERG; or KCNH2) encodes the voltage-gated potassium channel underlying I Kr , a repolarizing current in the heart. Mutations in KCNH2 or pharmacological agents that reduce I Kr slow action potential (AP) repolarization and can trigger cardiac arrhythmias associated with long QT syndrome. Two channel-forming subunits encoded by KCNH2 (hERG 1a and 1b) are expressed in cardiac tissue. In heterologous expression systems, these subunits avidly coassemble and exhibit biophysical and pharmacological properties distinct from those of homomeric hERG 1a channels. Despite these findings, adoption of hERG 1a/1b heteromeric channels as a model for cardiac I Kr has been hampered by the lack of evidence for a direct functional role for the 1b subunit in native tissue. In this study, we measured I Kr and APs at physiological temperature in cardiomyocytes derived from human induced pluripotent stem cells (iPSC-CMs). We found that specific knockdown of the 1b subunit using shRNA caused reductions in 1b mRNA, 1b protein levels, and I Kr magnitude by roughly one-half. AP duration was increased and AP variability was enhanced relative to controls. Early afterdepolarizations, considered cellular substrates for arrhythmia, were also observed in cells with reduced 1b expression. Similar behavior was elicited when channels were effectively converted from heteromers to 1a homomers by expressing a fragment corresponding to the 1a-specific N-terminal Per-Arnt-Sim domain, which is omitted from hERG 1b by alternate transcription. These findings establish that hERG 1b is critical for normal repolarization and that loss of 1b is proarrhythmic in human cardiac cells.T he human ether-à-go-go-related gene (hERG; or KCNH2) encodes the voltage-gated potassium channel underlying a native cardiac current known as I Kr (1, 2). Mutations in the gene or drugs that block the channel can diminish I Kr and slow ventricular repolarization, thus prolonging the QT interval on the surface electrocardiogram and causing long QT syndrome (LQTS) (1-3). Individuals with LQTS have an increased risk for torsades de pointes arrhythmia, syncope, and sudden cardiac death. Since 2005, a cell-based assay expressing the hERG 1a isoform as a proxy for I Kr has been the cornerstone of drug safety screening for new drug development (4). Thus, whether current drug safety assays accurately represent the native channel is of paramount importance.The first studies of heterologously expressed hERG channels described biophysical (1, 2) and pharmacological (2, 5) properties uniquely characteristic of cardiac I Kr . Subsequently, alternate transcripts of KCNH2 in mouse and human heart were shown to encode two subunits: 1a (the original isolate) and 1b (6, 7). In the hERG 1b transcript, an alternate 5′ exon replaces 1a exons 1-5, resulting in a shorter, unique N terminus that lacks a Per-Arnt-Sim (PAS) domain (also known as the ether-à-go-go domain) (8, 9). In heterologous systems, hERG 1b subunits avidly associate with hERG 1a but fail as homomer...

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Section: Discussionmentioning

confidence: 99%

hERG 1b is critical for human cardiac repolarization

Jones¹,

Liu²,

Vaidyanathan

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

108

View full text Add to dashboard Cite

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“…However, although K v 11.1 block is a discrete indicator of arrhythmogenicity, the former does not imply the latter, as demonstrated for verapamil (Liang et al , 2013) and ranolazine (Hancox et al , 2008), with a significant chance of generating false‐positive or false‐negative outcomes. Moreover, no information is obtained on the contractile behaviour or temporal dispersion of repolarization; the latter, with a crucial role in the genesis of lethal arrhythmia (Townsend and Brown, 2013), cannot be derived from simplified in vitro models, which further limits the predictivity of these approaches.…”

Section: Cardiotoxicity Screening Methodologiesmentioning

confidence: 99%

Integrating cardiomyocytes from human pluripotent stem cells in safety pharmacology: has the time come?

Sala

Bellin

Mummery

2016

British J Pharmacology

107

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Cardiotoxicity is a severe side effect of drugs that induce structural or electrophysiological changes in heart muscle cells. As a result, the heart undergoes failure and potentially lethal arrhythmias. It is still a major reason for drug failure in preclinical and clinical phases of drug discovery. Current methods for predicting cardiotoxicity are based on guidelines that combine electrophysiological analysis of cell lines expressing ion channels ectopically in vitro with animal models and clinical trials. Although no new cases of drugs linked to lethal arrhythmias have been reported since the introduction of these guidelines in 2005, their limited predictive power likely means that potentially valuable drugs may not reach clinical practice. Human pluripotent stem cell‐derived cardiomyocytes (hPSC‐CMs) are now emerging as potentially more predictive alternatives, particularly for the early phases of preclinical research. However, these cells are phenotypically immature and culture and assay methods not standardized, which could be a hurdle to the development of predictive computational models and their implementation into the drug discovery pipeline, in contrast to the ambitions of the comprehensive pro‐arrhythmia in vitro assay (CiPA) initiative. Here, we review present and future preclinical cardiotoxicity screening and suggest possible hPSC‐CM‐based strategies that may help to move the field forward. Coordinated efforts by basic scientists, companies and hPSC banks to standardize experimental conditions for generating reliable and reproducible safety indices will be helpful not only for cardiotoxicity prediction but also for precision medicine.Linked ArticlesThis article is part of a themed section on New Insights into Cardiotoxicity Caused by Chemotherapeutic Agents. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v174.21/issuetoc

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“…Given the cost of drug development, the ability to screen drugs for any condition against possible cardiac toxicity is a clear example of the effi cacy and cost -effectiveness of such developments [12]. The diffi culties in deriving homogeneous populations from pluripotent stem cells are acknowledged, but solvable.…”

Section: Introductionmentioning

confidence: 99%

National Institutes of Health Center for Regenerative Medicine: Putting Science into Practice

Rao

2013

Stem Cells and Development

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Proactive leadership rather than reactive stewardshipThe NIH is the premier source of medical funding in the United States and, as such, its allocation efforts strongly infl uence current and future strategies of development in regenerative medicine. I contend that the NIH has and will continue to anticipate the dizzying pace with which research discoveries are being advocated for and indeed, in some cases, research and leads the efforts at translating that science into clinical practice in the United States. The dollar amount invested specifi cally into stem cell research approximates a billion dollars a year (www.nih.gov). However, NIH support of regenerative medicine is not limited simply providing research dollars. To enable the translational efforts in the United States, the NIH needs to not only anticipate how scientifi c breakthroughs can inform regenerative medicine but also proactively lead reconceptualization of how therapeutic development can be supported and implemented.The National Institutes of Health and its role in putting medical science into practiceThe National Institutes of Health (NIH) plays a pivotal role in funding medical ABSTRACTThe fi eld of regenerative medicine has been revolutionized by breakthroughs in stem cell biology, gene engineering, and whole -genome sequencing. These advances are not only scientifi c or medical but have also advanced how we conceptualize regenerative medicine. The progenitive research that proceeded as well as a substantial part of the funding that supported these discoveries were provided by the National Institutes of Health (NIH). Now, perhaps more than ever, the NIH has a vital role to play in the translation of science into clinical practice. The NIH is uniquely positioned to coordinate interactions between the different institutes and other arms of the government, as well as international organizations. Efforts of researchers in the United States both within and without the NIH are supported by a number of mechanisms, including specialized workshops, and the support of developing small -scale industry. Additionally, the NIH has stepped up to provide necessary infrastructure in areas of regenerative medicine where the medical need might be apparent but might be currently infeasible or unattractive to private -sector investment. This article will discuss these perhaps lesser -known activities of the NIH, which I believe have continued and will continue to contribute to the role of stem cell research in translating science into regenerative medicine. The dollar amount invested specifi cally into stem cell research by the NIH approximates a billion dollars a year.

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Drug Screening Using a Library of Human Induced Pluripotent Stem Cell–Derived Cardiomyocytes Reveals Disease-Specific Patterns of Cardiotoxicity

Cited by 494 publications

References 46 publications

hERG 1b is critical for human cardiac repolarization

hERG 1b is critical for human cardiac repolarization

Integrating cardiomyocytes from human pluripotent stem cells in safety pharmacology: has the time come?

National Institutes of Health Center for Regenerative Medicine: Putting Science into Practice

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