Cardiac Differentiation of Adipose Tissue-Derived Stem Cells by Light Stimulation

描述（由申请人提供）：心肌梗死表现为肌纤维丧失，最终无法再生肌肉组织。基于细胞的疗法可以恢复缺血区域，并且脂肪组织可以为这种疗法提供容易的细胞来源。我们先前已经表明，人体脂肪组织含有细胞，有潜力分化成心血管系统。现有分化方案的主要缺点之一是脂肪组织来源的干细胞（ASC）的产量非常低，并且生化刺激产生随机空间组织和电不同步的心肌细胞集落。因此，显然需要新的方法来理解和操纵分化过程，以实现更高产量的同步心肌细胞，可以有效地移植到受损的心脏内。为了实现这一目标，我们将利用一种新的光遗传学系统，该系统最近已被证明可以在一系列时间尺度上实现可调的神经元激活和抑制。具体而言，在我们的目标中，我们提出：1）鉴定心包脂肪组织（PTA）内心脏祖细胞的特定亚群。基于上述初步数据，我们的工作假设是PTA含有充足的多能细胞来源。2)确定光遗传学在调节人脂肪干细胞（hASC）分化为心血管细胞中的作用。基于我们的初步数据，我们再次假设，连续膜电位去极化促进hASC定型以促进CM的产生，以及3）评估移植的电同步hASC对缺血性心肌再生的贡献。已经显示移植的hASCs-CM改善心脏功能，在此我们假设同步hASCs-CM可以显示更高的植入率并且直接有助于收缩功能。这些实验将有助于我们了解ASCs的分化机制，并为自体干细胞移植治疗缺血性心肌病提供依据。 公共卫生相关性：该项目与NIH的使命相关，因为缺血性心肌病患者将受益于注射的分化心肌细胞可以并入心肌的治疗。我们已经证明血管周围脂肪基质细胞具有分化为心肌细胞的潜力，并且使用我们的新方法，我们可以刺激细胞将其重定向为心肌细胞。因此，我们建议表征和增强这些细胞的潜力，并探索它们如何使用光遗传学刺激帮助心肌再生。本研究为自体组织作为终末期缺血性心脏病治疗的细胞来源提供了依据。

DESCRIPTION (provided by applicant): Myocardial infarction shows a loss of muscle fibers with ultimate failure to regenerate muscle tissue. Cell based therapies may restore ischemic areas and adipose tissue can provide a facile source of cells for such therapy. We have previously shown that human adipose tissue contains cells which have the potential to differentiate into cardiovascular linage. One of the main disadvantages of the existing differentiation protocols is that yields of Adipose Tissue-Derived Stem Cells (ASCs) have been very low and biochemical stimulation produces random spatial organization and electrically non-synchronization cardiomyocytes colonies. Therefore there is an obvious necessity for new approaches to understand and manipulate the differentiation process in order to achieve higher yields of synchronous cardiomyocytes that can engraft effectively within the damaged heart. To approach this goal we will utilize a novel optogentics system that has been recently shown to allow tunable neuronal activation and inhibition over a range of timescales. Specifically, in our aims we propose to: 1) Identify specific sub-populations of cardiac progenitor cells within Pericardial Adipose Tissue (PTA). Based on preliminary data referred to above, our working hypothesis is that PTA contains an ample source of pluripotent cells. 2) Determine the role of opto-genetics, in regulating human adipose stem cells (hASCs) differentiation to cardiovascular cells. We postulate again on the basis of our preliminary data, that continuous membrane potential depolarization promote hASCs commitment to facilitate the generation of CMs and 3) Evaluate the contribution of transplanted electrically synchronous hASCs to ischemic myocardial regeneration. It has been shown that transplanted hASCs-CMs improve cardiac function, here we hypothesize that synchronous hASCs-CMs can show higher rate of engraftment and directly contribute to the systolic function. These experiments should help us understand the mechanisms of ASCs differentiation and provide the basis for autologous stem cell transplantation for the treatment of ischemic cardiomyopathy. PUBLIC HEALTH RELEVANCE: The project is relevant to the NIH mission because patients with ischemic cardiomyopathy will benefit from therapy whereby injected differentiated cardiomyocytes can be incorporated into the heart muscle. We have already shown that perivascular adipose stromal cells have a potential in differentiating into cardiomyocytes and that using our novel approach we can stimulate cells to redirect them into cardiac linage cells. Hereby we propose to characterize and enhance the potential of these cells and explore how they can help in cardiac muscle regeneration using optogenetic stimulation. This research could provide a basis for autogenous tissue being used as a source of cells for the treatment of end-stage ischemic heart disease.