Electrical stimulation of human CPCs

项目总结/摘要：每120名出生儿童中就有近1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心力衰竭（HF）。的 心血管再生医学作为儿科HF的潜在治疗策略的出现， 为治疗提供了新的途径。虽然干细胞疗法主要在成年人中进行试验，但在成年人中相对未经测试。 儿科人群。因此，开发新的方法来探索再生潜力是至关重要的。 这些细胞来改善儿科人群的治疗干预。基于干细胞的疗法 对成人的几种心血管疾病显示出有益的作用， 心肌细胞（CPC）是一种在心肌中发现的祖细胞，在成人的临床试验中取得了早期成功。 由于能够在手术过程中容易地从CHD患者中分离这些细胞，因此它们在CHD患者中增殖的能力是非常重要的。 培养，和他们的易于操作，儿科CPC作为一个理想的细胞群体， 药然而，先前的研究表明，除非这些细胞在很小的年龄（<1 月），这些细胞的治疗功效降低。绝大多数的病人 手术治疗>1个月大的CHD，寻找新的方法来增强这些细胞的再生潜力， 克服干细胞治疗的这一关键障碍，并允许自体和异体治疗 儿童和成人的选择。已经表明，成年CPC的再生潜力可以是 通过离体操作增强。电刺激（ES）是一种已知的增强心源性的治疗方法， 干细胞是干细胞的一个重要组成部分，但其机制尚未确定。我们公布的数据显示 儿童CPC（从1-5岁的患者中分离）对ES的反应是通过启动钙 (Ca2+）振荡，使其成为用于通过离体ES操作的理想细胞群体。此外，我们的 数据表明ES增强了儿科CPC在体内的功能和保留。的目的 这项研究的目的是检测儿童CPC对ES的保护/再生能力。我们的目标是 表征ES治疗的儿科CPC释放的旁分泌因子，并确定其对心脏的影响。 细胞，表征增强的粘附/保留的机制，并最终确定ES是否增强其粘附/保留。 在体内发挥作用。该项目的成功完成将提供机制， 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 药该项目还直接解决了儿科HF干细胞治疗的关键障碍， 使用预处理的CPC提供自体和同种异体治疗。此外，我们的初步数据显示， 提示我们的研究结果可能适用于其他心脏适应症，如心肌梗死， 梗塞我们的研究结果将对所有基于细胞的疗法产生影响，并为以下方面提供有价值的见解： 从儿童到成人，增强干/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.