Enhancing the effectiveness of human cardiac stem cell therapy

项目摘要 我们的长期目标是开发可行的策略，以提高心脏干/祖细胞的疗效， 细胞（CPC）治疗，这是严重受阻的细胞来源有限的贡献，以产生新的 心肌细胞、细胞老化和供体细胞存活率差。我们以前证明，预处理 具有钴原卟啉（CoPP）人CPC表现出显著的改善细胞增殖的有益效果， 存活率和心脏功能。然而，单一策略对改善股骨柄的益处不足 细胞治疗效率。因此，本申请中的总体目标是测试组合的三个 小分子化合物，以增强新的CPC亚群的心肌修复效果， 低线粒体膜电位的人CPCs。在强大的初步数据的指导下，我们 中心假设是，老化的hCPCs已经损害了心脏再生潜力， 用三种小分子化合物组合使衰老的人低-hCPC恢复活力， 通过靶向多个衰老信号通路，增强体外再生潜力， 干细胞治疗免疫缺陷小鼠缺血性心脏病有效性 心肌梗死这一假设将通过追求三个具体目标来解决：我们将首先检验 用三种小分子化合物（3SMC）的复合物进行衰老的返老还童是否会降低 基于a）细胞衰老表型，B） 增殖，c）细胞存活，d）细胞因子释放，和e）内皮和心肌分化，通过 与媒介物处理的人巨噬细胞mlow-hCPCs和来自儿科患者的人巨噬细胞mlow-hCPCs进行比较（目的1）;我们的 目的二是了解衰老的Δ HCMLOW-hCPCs返老还童的分子和细胞机制 通过靶向衰老相关的信号通路与组合的3SMC结合。最后，我们将测试 3SMC处理的人低-hCPCs的移植是否会导致a）体内的更大改善 hCPC存活、增殖和分化; B）内源性小鼠心脏再生;和c）心脏 结构和功能在免疫缺陷小鼠心肌梗死（MI）模型（目的3）。该方法是 创新，在申请人的意见，因为它预计将设置一个新的研究里程碑， 小分子化合物的复合物，以增强新的亚组的治疗效果， 具有低线粒体膜电位的hCPC。这项研究意义重大，因为知识 从这些研究中获得的信息将为促进治疗的策略提供重要的启示。 移植后血小板mlow-hCPCs的有效性及其在临床修复梗死的有效性 用于缺血性心脏病患者的细胞治疗。

PROJECT SUMMARY Our long-term goal is to develop feasible strategies toward improving the efficacy of cardiac stem/progenitor cell (CPC) therapy, which is severely blocked by the cell source with limited contribution to produce new cardiomyocytes, cell aging, and poor donor cell survival. We previously demonstrated that preconditioning human CPCs with cobalt protoporphyrin (CoPP) exhibited significant beneficial effects for improving cell survival and cardiac function. However, singular strategy has shown insufficient benefit for improving the stem cell therapeutic efficiency. Therefore, the overall objective in this application is to test the compositional three small-molecule compounds to enhance the effectiveness of myocardial repair by a new subset of CPCs with low mitochondrial membrane potential, named as ∆ψmlow-hCPCs. Guided by the strong preliminary data, our central hypothesis is that aging ∆ψmlow-hCPCs have compromised cardiac regenerative potential, while rejuvenating aging ∆ψmlow-hCPCs with combined three small-molecule compounds will restore their cardiac regenerative potential in vitro through targeting multiple senescence signal pathways, and enhance the effectiveness of stem cell therapy for ischemic heart disease in an immuno-deficient murine model following myocardial infarction. This hypothesis will be addressed by pursuing three specific aims: we will first test whether the rejuvenation with a composite of three small-molecule compounds (3SMCs) for aging ∆ψmlow- hCPCs will increase their cardiac regenerative capability, on the basis of a) cell senescent phenotype, b) proliferation, c) cell survival, d) cytokine release, and e) endothelial and cardiomyogenic differentiation, through comparing with the vehicle-treated ∆ψmlow-hCPCs, and with ∆ψmlow-hCPCs from pediatric patients (Aim 1); Our Aim 2 is to understand the underlying molecular and cellular mechanisms of rejuvenating aging Δψmlow-hCPCs with combined 3SMCs by targeting the senescence-associated signaling pathways. Finally, we will test whether the transplantation of 3SMCs-treated ∆ψmlow-hCPCs will result in greater improvement of a) in vivo hCPC survival, proliferation, and differentiation; b) endogenous mouse cardiac regeneration; and c) cardiac structure and function in an immune-deficient mouse myocardial infarction (MI) model (Aim 3). The approach is innovative, in the applicant’s opinion, because it is expected to set a new research milestone on applying a composite of small-molecule compounds to enhance the therapeutic effectiveness of the novel subset of hCPCs with low mitochondrial membrane potential. The proposed research is significant, since knowledge gained from these studies will provide the essential implications on the strategies to promote the therapeutic effectiveness of ∆ψmlow-hCPCs after transplantation and, therefore, their efficacy in clinical repairing infarcted myocardium for the cell therapy of patients with ischemic heart disease.