Engineering of human cardiac muscle electromechanically matured to an adult-like phenotype

Ronaldson-Bouchard, Kacey; Yeager, Keith; Teles, Diogo; Chen, Timothy; Ma, Stephen; Song, LouJin; Morikawa, Kumi; Wobma, Holly; Vasciaveo, Alessandro; Ruiz, Edward C.; Yazawa, Masayuki; Vunjak‐Novakovic, Gordana

doi:10.1038/s41596-019-0189-8

Cited by 113 publications

(122 citation statements)

References 33 publications

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“…As for Ca 2+ handling, which is of central importance to excitation-contraction coupling, such related gene products as PLN and SERCA were upregulated, leading to improved calcium handling and subsequent contractility (Keung et al, 2016;Li et al, 2019). Other groups have reported similar findings where electrical pacing in hiPSC-CM tissue constructs promoted contractile force by ∼20-fold along with ∼2-and ∼1.5-fold increases in protein expression of SERCA and RYR, respectively, when compared to untreated controls (Ruan et al, 2016) or sensitivity toward isoproterenol, as indicated by calcium imaging (Ronaldson-Bouchard et al, 2019). Another finding of this study is the identification of the downregulation of the TGF-β signaling pathway induced by T3-EC combinatorial treatment.…”

Section: Discussionmentioning

confidence: 65%

“…Over the past decade or so, significant progress has been made in ventricular (V) specification to obtain high-purity VCMs (Ebert et al, 2012;Spater et al, 2014;Weng et al, 2014) for cardiac tissue engineering (Tallawi et al, 2015;Kolanowski et al, 2017;Wong et al, 2019). However, human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) possess immature fetal-like properties (Lieu et al, 2013;Robertson et al, 2013;Poon et al, 2015;Keung et al, 2016;Kadota et al, 2017;Ronaldson-Bouchard et al, 2019). Electrophysiologically, hPSC-CMs spontaneously fire action potentials (AP) with slower kinetics, starkly contrasting their quiescent-yet-excitable adult counterparts (Lieu et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

“…For instance, triiodothyronine (T3), the active form of the thyroid hormone in humans, is crucial for normal cardiac development (Klein and Danzi, 2007) and promotes the expression of a wide range of calcium handling and contractile proteins in murine and human CMs (Danzi and Klein, 2002;Lee et al, 2010;Yang et al, 2014b). Electrical conditioning (EC), including chronic electrical field pacing and stimulation-induced active contraction, upregulates important ion channel and calcium handling transcripts, thereby improving the structural alignment and promoting contractility of hPSC-derived cardiac tissue constructs (Lieu et al, 2013;Ruan et al, 2016;Ronaldson-Bouchard et al, 2019). Although these different biological and environmental cues were identified (Robertson et al, 2013;Yang et al, 2014a;Denning et al, 2016), a combinatorial approach to synergistically promote electrophysiological and contractile maturation of hPSC-CMs has not been developed.…”

Section: Introductionmentioning

confidence: 99%

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Combinatorial Treatment of Human Cardiac Engineered Tissues With Biomimetic Cues Induces Functional Maturation as Revealed by Optical Mapping of Action Potentials and Calcium Transients

Wong

Geng

et al. 2020

Front. Physiol.

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Although biomimetic stimuli, such as microgroove-induced alignment (µ), triiodothyronine (T3) induction, and electrical conditioning (EC), have been reported to promote maturation of human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs), a systematic examination of their combinatorial effects on engineered cardiac tissue constructs and the underlying molecular pathways has not been reported. Herein, human embryonic stem cell-derived ventricular cardiomyocytes (hESC-VCMs) were used to generate a micro-patterned human ventricular cardiac anisotropic sheets (hvCAS) for studying the physiological effects of combinatorial treatments by a range of functional, calcium (Ca 2+)-handling, and molecular analyses. High-resolution optical mapping showed that combined µ-T3-EC treatment of hvCAS increased the conduction velocity, anisotropic ratio, and proportion of mature quiescent-yet-excitable preparations by 2. 3-, 1. 8-, and 5-fold (>70%), respectively. Such electrophysiological changes could be attributed to an increase in inward sodium current density and a decrease in funny current densities, which is consistent with the observed upand downregulated SCN1B and HCN2/4 transcripts, respectively. Furthermore, Ca 2+-handling transcripts encoding for phospholamban (PLN) and sarco/endoplasmic reticulum Ca 2+-ATPase (SERCA) were upregulated, and this led to faster upstroke and decay kinetics of Ca 2+-transients. RNA-sequencing and pathway mapping of T3-EC-treated hvCAS revealed that the TGF-β signaling was downregulated; the TGF-β receptor agonist and antagonist TGF-β1 and SB431542 partially reversed T3-EC induced quiescence and reduced spontaneous contractions, respectively. Taken

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Section: Discussionmentioning

confidence: 65%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Combinatorial Treatment of Human Cardiac Engineered Tissues With Biomimetic Cues Induces Functional Maturation as Revealed by Optical Mapping of Action Potentials and Calcium Transients

Wong

Geng

et al. 2020

Front. Physiol.

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“…Strategies to achieve functional and structural maturation as well as the production of specific cardiac subtypes will enable more comprehensive studies of disease modeling and will widen the potential applications of hiPSC-CMs in preclinical studies. New technologies including human organoids and engineered human tissue mimicking the 3D structure and function of native cardiac tissue appear to be viable alternatives to currently used cell and animal models [161,162,[179][180][181][182][183][184]. Native-like in vitro environments obtained from mixing various cell populations and suitable biomaterials may provide the means to generate a mature electrophysiological phenotype, currently lacking in 2D cultures [161,185].…”

Section: Resultsmentioning

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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“…The dyn-EHT platform builds off of previous work from our laboratory and other investigators that have used primary, hESC and hiPSC derived cardiomyocytes to engineer functional cardiac tissues in vitro. Indeed, it is critical to acknowledge the important contributions to the field made by researchers using EHTs to model development and disease, and implementing more physiologic-like electromechanical conditioning to drive maturation (14,(16)(17)20,23,(25)(26)28,40,(61)(62)(63)(64). However, these 2D and 3D EHTs are generally isometrically constrained, which means they are unable to undergo significant fractional shortening or be stretched by preload.…”

Section: Discussionmentioning

confidence: 99%

Dynamic Loading of Human Engineered Heart Tissue Enhances Contractile Function and Drives Desmosome-linked Disease Phenotype

Bliley

Vermeer

Duffy

et al. 2020

Preprint

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The role mechanical forces play in shaping the structure and function of the heart is critical to understanding heart formation and the etiology of disease but is challenging to study in patients. Engineered heart tissues (EHTs) incorporating human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes have the potential to provide insight into these adaptive and maladaptive changes in the heart. However, most EHT systems are unable to model both preload (stretch during chamber filling) and afterload (pressure the heart must work against to eject blood). Here, we have developed a new dynamic EHT (dyn-EHT) model that enables us to tune preload and have unconstrained fractional shortening of >10%.To do this, 3D EHTs are integrated with an elastic polydimethylsiloxane (PDMS) strip that provides mechanical pre-and afterload to the tissue in addition to enabling contractile force measurements based on strip bending. Our results demonstrate in wild-type EHTs that dynamic loading is beneficial based on the magnitude of the forces, leading to improved alignment, conduction velocity, and contractility. For disease modeling, we use hiPSC-derived cardiomyocytes from a patient with arrhythmogenic cardiomyopathy (ACM) due to mutations in desmoplakin. We demonstrate that manifestation of this desmosome-linked disease state requires the dyn-EHT conditioning and that it cannot be induced using 2D or standard 3D EHT approaches. Thus, dynamic loading strategy is necessary to provoke a disease phenotype (diastolic lengthening, reduction of desmosome counts, and reduced contractility), which are akin to primary endpoints of clinical disease, such as chamber thinning and reduced cardiac output.Studying the effects of mechanical loading on adaptive and maladaptive changes in heart structure and function is challenging in human patients, driving the need to develop new approaches. Animals have been used to model a wide range of human cardiovascular disease states, but there are inherent differences in physiology, gene expression and pharmacokinetics that limit the ability to replicate complex pathology (6). In vitro 2-dimensional (2D) culture of cardiomyocytes on microengineered surfaces have been used to study structure-function relationships and to create region-specific ventricular myocardium (7,8) Muscular thin films (MTFs), consisting of cells cultured on a flexible film, have enabled these 2D platforms to measure contractility (9,10), and combined with patient-specific human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes can model cardiomyopathies such as Barth's syndrome (11). However, 2D cardiomyocytes are adhered to a surface (12) and thus do not have the same mechanical cell-cell coupling as found in the native heart (13). To address this, researchers have developed more physiologically-relevant 3D engineered heart tissues (EHTs) in the form papillary muscle-like linear bundles (14-18) myocardium-like sheets (19,20), and ventricle-like chambers (21,22). Incorporating human induced pluripotent stem cell (hi...

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Engineering of human cardiac muscle electromechanically matured to an adult-like phenotype

Cited by 113 publications

References 33 publications

Combinatorial Treatment of Human Cardiac Engineered Tissues With Biomimetic Cues Induces Functional Maturation as Revealed by Optical Mapping of Action Potentials and Calcium Transients

Combinatorial Treatment of Human Cardiac Engineered Tissues With Biomimetic Cues Induces Functional Maturation as Revealed by Optical Mapping of Action Potentials and Calcium Transients

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

Dynamic Loading of Human Engineered Heart Tissue Enhances Contractile Function and Drives Desmosome-linked Disease Phenotype

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