San Diego Heart Research Institute, San Diego State University, 5500 Campanile Drive, San Diego, CA, 92182, USA.
Sharp Memorial Hospital, San Diego, CA, 92123, USA.
J Physiol. 2017 Dec 1;595(23):7135-7148. doi: 10.1113/JP274980. Epub 2017 Nov 9.
Autologous cardiac progenitor cell (CPC) therapy is a promising approach for treatment of heart failure (HF). There is an unmet need to identify inherent deficits in aged/diseased human CPCs (hCPCs) derived from HF patients in the attempts to augment their regenerative capacity prior to use in the clinical setting. Here we report significant functional correlations between phenotypic properties of hCPCs isolated from cardiac biopsies of HF patients, clinical parameters of patients and expression of the P2Y purinergic receptor (P2Y R), a crucial detector for extracellular UDP-sugars released during injury/stress. P2Y R is downregulated in hCPCs derived from HF patients with lower ejection fraction or diagnosed with diabetes. Augmenting P2Y R expression levels in aged/diseased hCPCs antagonizes senescence and improves functional responses. This study introduces purinergic signalling modulation as a potential strategy to rejuvenate and improve phenotypic characteristics of aged/functionally compromised hCPCs prior to transplantation in HF patients.
Autologous cardiac progenitor cell therapy is a promising alternative approach to current inefficient therapies for heart failure (HF). However, ex vivo expansion and pharmacological/genetic modification of human cardiac progenitor cells (hCPCs) are necessary interventions to rejuvenate aged/diseased cells and improve their regenerative capacities. This study was designed to assess the potential of improving hCPC functional capacity by targeting the P2Y purinergic receptor (P2Y R), which has been previously reported to induce regenerative and anti-senescence responses in a variety of experimental models. c-Kit hCPCs were isolated from cardiac biopsies of multiple HF patients undergoing left ventricular assist device implantation surgery. Significant correlations existed between the expression of P2Y R in hCPCs and clinical parameters of HF patients. P2Y R was downregulated in hCPCs derived from patients with a relatively lower ejection fraction and patients diagnosed with diabetes. hCPC lines with lower P2Y R expression did not respond to P2Y R agonist UDP-glucose (UDP-Glu) while hCPCs with higher P2Y R expression showed enhanced proliferation in response to UDP-Glu stimulation. Mechanistically, UDP-Glu stimulation enhanced the activation of canonical growth signalling pathways ERK1/2 and AKT. Restoring P2Y R expression levels in functionally compromised hCPCs via lentiviral-mediated overexpression improved proliferation, migration and survival under stress stimuli. Additionally, P2Y R overexpression reversed senescence-associated morphology and reduced levels of molecular markers of senescence p16 , p53, p21 and mitochondrial reactive oxygen species. Findings from this study unveil novel biological roles of the UDP-sugar receptor P2Y in hCPCs and suggest purinergic signalling modulation as a promising strategy to improve phenotypic properties of functionally impaired hCPCs.
自体心脏祖细胞（CPC）疗法是治疗心力衰竭（HF）的一种很有前途的方法。在尝试增强其再生能力之前，需要识别从 HF 患者中分离得到的衰老/患病的人源 CPC（hCPC）的固有缺陷，以便将其用于临床环境。在这里，我们报告了从 HF 患者的心脏活检中分离出的 hCPC 的表型特性与患者的临床参数之间存在显著的功能相关性，以及 P2Y 嘌呤能受体（P2Y R）的表达，P2Y R 是损伤/应激期间释放的细胞外 UDP-糖的关键检测器。P2Y R 在 HF 患者中表达下调，其射血分数较低或被诊断患有糖尿病。在衰老/患病的 hCPC 中，增强 P2Y R 的表达水平可拮抗衰老并改善功能反应。这项研究介绍了嘌呤能信号转导调节作为一种潜在策略，可在将其移植到 HF 患者之前，恢复和改善衰老/功能受损 hCPC 的表型特征。
自体心脏祖细胞疗法是目前治疗心力衰竭（HF）的低效疗法的一种很有前途的替代方法。然而，体外扩增和人源心脏祖细胞（hCPC）的药理学/遗传学修饰是恢复衰老/患病细胞并提高其再生能力所必需的干预措施。本研究旨在通过靶向 P2Y 嘌呤能受体（P2Y R）来评估改善 hCPC 功能能力的潜力，先前的研究表明 P2Y R 在多种实验模型中可诱导再生和抗衰老反应。从接受左心室辅助装置植入手术的多位 HF 患者的心脏活检中分离出 c-Kit hCPC。hCPC 中 P2Y R 的表达与 HF 患者的临床参数之间存在显著相关性。P2Y R 在射血分数相对较低的患者和被诊断患有糖尿病的患者来源的 hCPC 中表达下调。表达较低 P2Y R 的 hCPC 系对 P2Y R 激动剂 UDP-葡萄糖（UDP-Glu）无反应，而表达较高 P2Y R 的 hCPC 系在 UDP-Glu 刺激下表现出增强的增殖反应。从机制上讲，UDP-Glu 刺激增强了经典生长信号通路 ERK1/2 和 AKT 的激活。通过慢病毒介导的过表达恢复功能受损 hCPC 中的 P2Y R 表达水平可改善应激刺激下的增殖、迁移和存活。此外，P2Y R 过表达逆转了衰老相关的形态并降低了衰老标志物 p16、p53、p21 和线粒体活性氧的水平。这项研究揭示了 UDP-糖受体 P2Y 在 hCPC 中的新生物学作用，并表明嘌呤能信号转导调节是改善功能受损 hCPC 表型特性的一种很有前途的策略。
