Electrical Stimulation of Human CPCs

项目摘要/摘要：每120名新生儿中就有一名患有先天性心脏病(CHD)。 虽然手术治疗提高了存活率，但这些儿童中的许多人继续发展为心力衰竭(HF)。这个 心血管再生医学的出现成为治疗儿童心力衰竭的潜在策略 为治疗提供了新的途径。虽然干细胞疗法主要在成人中试验，但在 儿科人口。因此，开发探索再生潜力的新方法是至关重要的。 这些细胞用于改善儿科人群的治疗干预。基于干细胞的疗法有 对成年人的几种心血管疾病和心脏来源的c-kit+祖细胞有有益的影响 细胞是在心肌中发现的一种前体细胞，在成年人的临床试验中取得了早期的成功。 由于能够在手术中很容易地从冠心病患者身上分离出这些细胞，它们在 培养和操作简单，儿科CPC是理想的再生细胞群 医药。然而，先前的研究表明，除非在非常年轻的时候提取这些细胞(&lt；1 月)，这些细胞的治疗效果就会减弱。绝大多数患者都在接受 对1个月大的先天性心脏病进行手术，寻找新的方法来增强这些细胞的再生潜力将 克服干细胞治疗的这一关键障碍，允许自体和异体治疗 儿童和成人的选择。已有研究表明，成体CPC的再生潜力可以是 通过体外操作来增强。电刺激(ES)是一种已知的增强心源性的治疗方法。 各种成体干细胞的潜能；然而，其机制仍未确定。我们公布的数据显示 儿科CPC(从1-5岁的患者中分离出来)通过启动钙离子对ES作出反应 (Ca~(2+))振荡使其成为体外ES操纵的理想细胞群。此外，我们的 数据表明，ES增强了儿科CPC的功能和在体内的保留。这样做的目的是 建议是检查儿科CPC对ES的保护/再生能力。我们的目标是 ES治疗的儿童CPC释放旁分泌因子的特征及其对心脏的影响 细胞，表征增强黏附/保持的机制，并最终确定ES是否增强了它们的 在体内发挥作用。该项目的成功完成将提供提高治疗效果的机制 心脏干细胞的功效，提供自体和同种异体治疗选择，并促进再生 医药。该项目还通过以下方式直接解决了儿童心衰干细胞疗法中的关键障碍 使用预处理的CPC提供自体和同种异体治疗。另外，我们的初步数据 提示我们的发现可能适用于其他心脏指征的更广泛的治疗，如心肌梗死 脑梗塞。我们的结果将对所有基于细胞的疗法产生影响，并提供有价值的见解 从儿童到成人提高干细胞/祖细胞治疗潜力的机制。

PROJECT SUMMARY/ABSTRACT: Nearly 1 in every 120 children born has a congenital heart defect (CHD). While surgical therapy has improved survival, many of these children go on to develop heart failure (HF). The emergence of cardiovascular regenerative medicine as a potential therapeutic strategy for pediatric HF has provided new avenues for treatment. While primarily tried in adults, stem cell therapy is relatively untested in the pediatric population. It is thus critical to develop novel methods of exploring the regenerative potential of these cells to improve therapeutic interventions in the pediatric population. Stem cell based therapies have shown beneficial effects on several cardiovascular diseases in adults, and cardiac-derived c-kit+ progenitor cells (CPCs), a progenitor cell found in the myocardium, have met with early success in a clinical trial in adults. Due to the ability to readily isolate these cells from CHD patients during surgery, their capacity to proliferate in culture, and their ease of manipulation, pediatric CPCs serve as an ideal cell population for regenerative medicine. However, previous studies have shown that unless these cells are extracted at a very young age (<1 month) the therapeutic efficacy of these cells is diminished. With the vast majority of patients undergoing surgery for CHDs >1 month old, finding novel ways to enhance the regenerative potential of these cells would overcome this critical barrier to stem cell therapy and allow for both autologous and allogeneic treatment options in children and adults. It has been shown that the regenerative potential of adult CPCs can be enhanced by ex vivo manipulation. Electrical stimulation (ES) is one treatment known to enhance cardiogenic potential of various adult stem cells; however, the mechanism remains undetermined. Our published data show that pediatric CPCs (isolated from patients between 1-5 years of age) respond to ES by initiating calcium (Ca2+) oscillations making them an ideal population of cells for manipulation by ex vivo ES. Additionally, our data indicate ES enhances both the function and the retention of pediatric CPCs in vivo. The objective of this proposal is to examine the protective/regenerative capacity of pediatric CPCs in response to ES. We aim to characterize the paracrine factors released by ES-treated pediatric CPCs and determine their effect on cardiac cells, characterize the mechanism of enhanced adhesion/retention, and finally determine if ES enhances their function in vivo. Successful completion of this project will provide mechanisms to enhance the therapeutic efficacy of cardiac stem cells, provide autologous and allogeneic treatment options, and advance regenerative medicine. This project also directly addresses a critical barrier in stem cell therapies for pediatric HF by providing autologous and allogeneic treatments using pre-conditioned CPCs. Additionally, our preliminary data suggest our findings may be applicable as a broader therapy for other cardiac indications such as myocardial infarction. Our results will have implications on all cell-based therapies and offer valuable insights into mechanisms to enhance the therapeutic potential of stem/progenitor cells from children to adults.