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Article: Phospholamban as a Crucial Determinant of the Inotropic Response of Human Pluripotent Stem Cell–Derived Ventricular Cardiomyocytes and Engineered 3-Dimensional Tissue Constructs

TitlePhospholamban as a Crucial Determinant of the Inotropic Response of Human Pluripotent Stem Cell–Derived Ventricular Cardiomyocytes and Engineered 3-Dimensional Tissue Constructs
Authors
Issue Date2015
Citation
Circulation: Arrhythmia and Electrophysiology, 2015, v. 8 n. 1, p. 193-202 How to Cite?
AbstractBACKGROUND: Human (h) embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) serve as a potential unlimited ex vivo source of cardiomyocytes (CMs). However, a well-accepted roadblock has been their immature phenotype. hESC/iPSC-derived ventricular (v) CMs and their engineered cardiac microtissues (hvCMTs) similarly displayed positive chronotropic but null inotropic responses to beta-adrenergic stimulation. Given that phospholamban (PLB) is robustly present in adult but poorly expressed in hESC/iPSC-vCMs and its defined biological role in beta-adrenergic signaling, we investigated the functional consequences of PLB expression in hESC/iPSC-vCMs and hvCMTs. METHODS AND RESULTS: First, we confirmed that PLB protein was differentially expressed in hESC (HES2, H9)- and iPSC-derived and adult vCMs. We then transduced hES2-vCMs with the recombinant adenoviruses (Ad) Ad-PLB or Ad-S16E-PLB to overexpress wild-type PLB or the pseudophosphorylated point-mutated variant, respectively. As anticipated from the inhibitory effect of unphosphorylated PLB on sarco/endoplasmic reticulum Ca2+-ATPase, Ad-PLB transduction significantly attenuated electrically evoked Ca2+ transient amplitude and prolonged the 50% decay time. Importantly, Ad-PLB-transduced hES2-vCMs uniquely responded to isoproterenol. Ad-S16E-PLB-transduced hES2-vCMs displayed an intermediate phenotype. The same trends were observed with H9- and iPSC-vCMs. Directionally, similar results were also seen with Ad-PLB-transduced and Ad-S16E-transduced hvCMTs. However, Ad-PLB altered neither the global transcriptome nor ICa,L, implicating a PLB-specific effect. CONCLUSIONS: Engineered upregulation of PLB expression in hESC/iPSC-vCMs restores a positive inotropic response to beta-adrenergic stimulation. These results not only provide a better mechanistic understanding of the immaturity of hESC/iPSC-vCMs but will also lead to improved disease models and transplantable prototypes with adult-like physiological responses.
Persistent Identifierhttp://hdl.handle.net/10722/208200
ISSN
2015 SCImago Journal Rankings: 2.844

 

DC FieldValueLanguage
dc.contributor.authorChen, Gen_US
dc.contributor.authorLI, Sen_US
dc.contributor.authorKarakikes, Ien_US
dc.contributor.authorRen, Len_US
dc.contributor.authorChow, MZYen_US
dc.contributor.authorChopra, Aen_US
dc.contributor.authorKeung, WWYen_US
dc.contributor.authorYan, Ben_US
dc.contributor.authorChan, CWYen_US
dc.contributor.authorCosta, KDen_US
dc.contributor.authorKong, CW-
dc.contributor.authorHajjar, RJ-
dc.contributor.authorChen, CS-
dc.contributor.authorLi, RA-
dc.date.accessioned2015-02-23T08:05:35Z-
dc.date.available2015-02-23T08:05:35Z-
dc.date.issued2015en_US
dc.identifier.citationCirculation: Arrhythmia and Electrophysiology, 2015, v. 8 n. 1, p. 193-202en_US
dc.identifier.issn1941-3084-
dc.identifier.urihttp://hdl.handle.net/10722/208200-
dc.description.abstractBACKGROUND: Human (h) embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) serve as a potential unlimited ex vivo source of cardiomyocytes (CMs). However, a well-accepted roadblock has been their immature phenotype. hESC/iPSC-derived ventricular (v) CMs and their engineered cardiac microtissues (hvCMTs) similarly displayed positive chronotropic but null inotropic responses to beta-adrenergic stimulation. Given that phospholamban (PLB) is robustly present in adult but poorly expressed in hESC/iPSC-vCMs and its defined biological role in beta-adrenergic signaling, we investigated the functional consequences of PLB expression in hESC/iPSC-vCMs and hvCMTs. METHODS AND RESULTS: First, we confirmed that PLB protein was differentially expressed in hESC (HES2, H9)- and iPSC-derived and adult vCMs. We then transduced hES2-vCMs with the recombinant adenoviruses (Ad) Ad-PLB or Ad-S16E-PLB to overexpress wild-type PLB or the pseudophosphorylated point-mutated variant, respectively. As anticipated from the inhibitory effect of unphosphorylated PLB on sarco/endoplasmic reticulum Ca2+-ATPase, Ad-PLB transduction significantly attenuated electrically evoked Ca2+ transient amplitude and prolonged the 50% decay time. Importantly, Ad-PLB-transduced hES2-vCMs uniquely responded to isoproterenol. Ad-S16E-PLB-transduced hES2-vCMs displayed an intermediate phenotype. The same trends were observed with H9- and iPSC-vCMs. Directionally, similar results were also seen with Ad-PLB-transduced and Ad-S16E-transduced hvCMTs. However, Ad-PLB altered neither the global transcriptome nor ICa,L, implicating a PLB-specific effect. CONCLUSIONS: Engineered upregulation of PLB expression in hESC/iPSC-vCMs restores a positive inotropic response to beta-adrenergic stimulation. These results not only provide a better mechanistic understanding of the immaturity of hESC/iPSC-vCMs but will also lead to improved disease models and transplantable prototypes with adult-like physiological responses.-
dc.languageengen_US
dc.relation.ispartofCirculation: Arrhythmia and Electrophysiologyen_US
dc.titlePhospholamban as a Crucial Determinant of the Inotropic Response of Human Pluripotent Stem Cell–Derived Ventricular Cardiomyocytes and Engineered 3-Dimensional Tissue Constructsen_US
dc.typeArticleen_US
dc.identifier.emailRen, L: renlh@hku.hken_US
dc.identifier.emailChow, MZY: mc1024@hku.hken_US
dc.identifier.emailKeung, WWY: wkeung@hku.hken_US
dc.identifier.emailYan, B: yanbin14@hku.hken_US
dc.identifier.emailChan, CWY: camchan@hku.hken_US
dc.identifier.authorityKeung, WWY=rp01887en_US
dc.identifier.authorityYan, B=rp01940en_US
dc.identifier.authorityChan, CWY=rp01311en_US
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1161/CIRCEP.114.002049en_US
dc.identifier.pmid25504561-
dc.identifier.scopuseid_2-s2.0-84923852701-
dc.identifier.hkuros242521en_US
dc.identifier.hkuros254311-

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