Phasic modulation of Wnt signaling enhances cardiac differentiation in human pluripotent stem cells by recapitulating developmental ontogeny

Abstract

Cardiomyocytes (CMs) derived from human pluripotent stem cells (hPSCs) offer immense value in studying cardiovascular regenerative medicine. However, intrinsic biases and differential responsiveness of hPSCs towards cardiac differentiation pose significant technical and logistic hurdles that hamper human cardiomyocyte studies. Tandem modulation of canonical and non-canonical Wnt signaling pathways may play a crucial role in cardiac development that can efficiently generate cardiomyocytes from pluripotent stem cells. Our Wnt signaling expression profiles revealed that phasic modulation of canonical/non-canonical axis enabled orderly recapitulation of cardiac developmental ontogeny. Moreover, evaluation of 8 hPSC lines showed marked commitment towards cardiac-mesoderm during the early phase of differentiation, with elevated levels of canonical Wnts (Wnt3 and 3a) and Mesp1. Whereas continued activation of canonical Wnts was counterproductive, its discrete inhibition during the later phase of cardiac differentiation was accompanied by significant up-regulation of non-canonical Wnt expression (Wnt5a and 11) and enhanced Nkx2.5(+) (up to 98%) populations. These Nkx2.5(+) populations transited to contracting cardiac troponin T-positive CMs with up to 80% efficiency. Our results suggest that timely modulation of Wnt pathways would transcend intrinsic differentiation biases of hPSCs to consistently generate functional CMs that could facilitate their scalable production for meaningful clinical translation towards personalized regenerative medicine.

Arsenic and the Cardiovascular System

“…However, reports have shown that there are inherent differences within various pluripotent stem cell lines that may affect their differentiation abilities to various lineages [143]. However, having efficient differentiation protocols could resolve these inherent differences [142,144].…”

“…Human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), through defined culture conditions, can be differentiated into various cell types, including cardiomyocytes [142]. Therefore, human ES-and iPSC-derived cardiomyocytes provide a more suitable alternative to animal models.…”

Human Pluripotent Stem Cells and Drug Discovery: A New Beginning

“…The FDA has thus recognised QT prolongation as an important variable which needs to be addressed during drug development while the International Conference of Harmonisation (ICH) Expert Working Group (EWG) for drug development has placed risk assessments for delayed ventricular repolarisation as standard procedure of preclinical evaluation of new chemical entities 24 . Cardiomyocytes differentiated from hPSCs open a new paradigm in regenerative medicine 25,26 , disease modelling 20,27 , drug discovery [18][19][20]28 and understanding developmental phenomena 17,29 . Similar to their adult counterparts, hPSC-derived cardiomyocytes express cardiac genes, structural proteins and ion channels 17,30 as well as exhibit a functional excitation-contraction coupling system which could be modulated through β1/β2-adrenergic and muscarinic receptor agonists 31 .…”

“…Cardiomyocytes differentiated from hPSCs open a new paradigm in regenerative medicine 25,26 , disease modelling 20,27 , drug discovery [18][19][20]28 and understanding developmental phenomena 17,29 . Similar to their adult counterparts, hPSC-derived cardiomyocytes express cardiac genes, structural proteins and ion channels 17,30 as well as exhibit a functional excitation-contraction coupling system which could be modulated through β1/β2-adrenergic and muscarinic receptor agonists 31 . Evaluation of electrophysiological properties could be performed on single cells (patch clamp) or clusters (multi-electrode analysis; MEA) which allow for studying action potential (AP) and field potential duration (FPD) respectively (apart from other parameters).…”

“…The ability of human pluripotent stem cells (hPSCs) to self-renew and differentiate into virtually any cell type of the three germ layers provides an unparalleled advantage over somatic cell test systems 12,13 . Their unlimited supply and reproducible differentiation [14][15][16][17] into desired cell types allows for unique opportunities in drug development 5, [18][19][20][21] . In this mini-review, we direct our focus on the advantages and disadvantages of utilising hPSC-derived cell types as drug discovery platforms with special emphasis on cardioand hepatotoxicity; forerunners for drug attrition as well as embryotoxicity.…”