P2Y ₂ Nucleotide Receptor Prompts Human Cardiac Progenitor Cell Activation by Modulating Hippo Signaling

Abstract

Rationale Autologous stem cell therapy using human c-Kit+ cardiac progenitor cells (hCPCs) is a promising therapeutic approach for treatment of heart failure (HF). However, hCPCs derived from aged HF patients with genetic predispositions and/or comorbidities of chronic diseases exhibit poor proliferative and migratory capabilities, which impairs overall reparative potential for injured myocardium. Therefore, empowering functionally compromised hCPCs with pro-regenerative molecules ex vivo is crucial for improving the therapeutic outcome in HF patients. Objective To improve hCPC proliferation and migration responses that are critical for regeneration by targeting pro-regenerative P2Y2 nucleotide receptor (P2Y2R) activated by extracellular ATP and UTP molecules released following injury/stress. Methods and Results c-Kit+ hCPCs were isolated from cardiac tissue of HF patients undergoing left ventricular assist device (LVAD) implantation surgery. Correlations between P2 nucleotide receptor expression and hCPC growth kinetics revealed downregulation of select P2 receptors, including P2Y2R, in slow-growing hCPCs compared to fast-growers. hCPC proliferation and migration significantly improved by overexpressing or stimulating P2Y2R. Mechanistically, P2Y2R-induced proliferation and migration were dependent upon activation of yes-associated protein (YAP), the downstream effector of Hippo signaling pathway. Conclusions Proliferation and migration of functionally impaired hCPCs are enhanced by P2Y2R-mediated YAP activation, revealing a novel link between extracellular nucleotides released during injury/stress and Hippo signaling, a central regulator of cardiac regeneration. Functional correlations exist between hCPC phenotypic properties and P2 purinergic receptor expression. Lack of P2Y2R and other crucial purinergic stress detectors could compromise hCPC responsiveness to presence of extracellular stress signals. These findings set the stage for subsequent studies to assess purinergic signaling modulation as a potential strategy to improve therapeutic outcome for use of hCPCs in HF patients.

Hippo Signaling Controls NLR Family Pyrin Domain Containing 3 Activation and Governs Immunoregulation of Mesenchymal Stem Cells in Mouse Liver Injury

“…The purinergic receptor signaling was crucial for the ATP‐mediated NLRP3 activation in allograft rejection . Interestingly, the purinergic receptors induced cell proliferation and migration in injured tissues by regulating YAP activation, suggesting that there is a crosstalk between purinergic receptors and the Hippo‐YAP pathway. Moreover, the hypoxia‐inducible factors regulated their target genes, leading to resistance to ischemia and controlling excessive inflammation, which may interact with YAP under hypoxic conditions .…”

Yes-associated protein (YAP) in pancreatic cancer: at the epicenter of a targetable signaling network associated with patient survival

“…GPCR signaling is one of the major upstream signals that regulate YAP/TAZ activation in a variety of systems, including PDAC 31 . In this context, the expression of genes encoding the lysophosphatidic acid (LPA) receptor GPR87 86 , the purinergic GPCR P2Y 2 R 87 and the GPCR agonist EDN2 (endothelin 2) 57 , a downstream target of YAP, are also associated with an unfavorable prognosis in PDAC. A recent independent study confirmed that overexpression of GPR87 is correlated with a poor prognosis in PDAC 88 .…”

Role of Hippo-YAP1/TAZ pathway and its crosstalk in cardiac biology

“…P2Y 2 receptor (P2Y 2 R) is a purinergic GPCR that regulates Gα q/11 -PKC signaling to control the flow of Ca 2+ and K + ions 90 . The P2Y 2 R-PKC axis activates YAP1 by inhibiting LATS1/2 and subsequently promote cell proliferation in human c-Kit + cardiac progenitor cells (hCPCs), which helps to rescue CM from damages 91 , 92 . Moreover, the GPCR agonist neurotensin attenuates YAP1 phosphorylation at Ser127/397 and augments YAP1 nuclear localization via PKD signaling 93 .…”