Rhesus Heart Regeneration With Embryonic Stem Cells

描述(由申请人提供)： 许多形式的心脏病的特征是心肌细胞的丧失，或者是细胞凋亡，或者是心肌梗死。因此，心脏细胞移植等策略可能在一系列心脏疾病的治疗方面提供显著的进步。各种供体细胞已经在动物模型的脑梗塞中进行了测试，其中最有希望的结果是成人干细胞。然而，胚胎干细胞可能为心肌再生提供最大的机会。这项建议的总体目标是利用非人灵长类胚胎干细胞再生梗塞心肌，作为迈向心脏细胞移植临床应用的积极一步。多学科方法将提供创新的方法，以解决将细胞移植引入临床的主要障碍。首先，我们假设，通过适当的启动，恒河猴ES细胞将能够移植到梗死的恒河猴心肌中，并再生出多种类型的典型心肌细胞。特定目标1将测试标记有荧光蛋白EGFP的供体ES细胞在恒河猴心肌梗死-再灌注模型中的植入情况。体外免疫组织化学将提供供体细胞再生细胞类型的信息，分离的心肌细胞研究将表征再生心肌细胞的细胞功能。特定目标2将开发使用磁共振成像(MRI)的新的成像应用，以跟踪标记有磁树突状分子的供体ES细胞，并提供心肌再生的结构和功能细节。接下来，我们假设梗死心肌的微环境具有丰富的信号分子集合，指导祖细胞募集和环境依赖的分化。在具体目标3中，我们将使用基于抗体的分析方法以及利用蛋白质组学技术和质谱仪，在脑梗塞后的静脉流出物中寻找这些信号分子。为了在受控的微环境中测试可能的信号分子，我们将开发一种微流控生物检测系统，使用微通道中培养的ES细胞。总体而言，这些开发方法有可能极大地促进心肌再生领域的发展，并使心脏细胞移植更接近临床现实。

DESCRIPTION (provided by applicant): Many forms of heart disease are characterized by a loss of myocytes either by apoptosis or infarction. Therefore, strategies such as cardiac cell transplantation may provide remarkable advances in therapy for a range of heart diseases. A variety of donor cells have been tested in animal models of infarction with the most promising results from adult stem cells. However, embryonic stem (ES) cells may provide the greatest opportunity for myocardial regeneration. The overall goal of this proposal is to utilize nonhuman primate embryonic stem (ES) cells to regenerate infarcted myocardium as an aggressive step toward clinical applications of cardiac cell transplantation. A multidisciplinary approach will provide innovative approaches to address major roadblocks in bringing cell transplantation to the clinic. First, we hypothesize that, with appropriate priming, rhesus ES cells will be able to engraft into the infarcted rhesus myocardium and regenerate multiple cell types typical of myocardium. Specific Aim 1 will test engraftment of donor ES cells labeled with the fluorescent protein EGFP in a rhesus myocardial infarction-reperfusion model. Ex vivo immunohistochemistry will provide information on the cell types regenerated from the donor cells, and isolated myocyte studies will characterize the cellular function of regenerated myocytes. Specific Aim 2 will develop novel imaging applications using Magnetic Resonance Imaging (MRI) to track donor ES cells labeled with magnetodendrimers as well as provide structural and functional details of myocardial regeneration. Next, we hypothesize that the microenvironment of the infarcted myocardium has a rich collection of signaling molecules directing progenitor cell recruitment and milieu-dependent differentiation. In Specific Aim 3, we will search for these signaling molecules in the venous effluent following infarction using antibody-based assays as well as utilizing proteomics technology with mass spectrometry. To test putative signaling molecules in a controlled microenvironment, we will develop a microfluidics bioassay system using cultured ES cells in microchannels. Overall, these developmental approaches have the potential to dramatically advance the field of myocardial regeneration and bring cardiac cell transplantation much closer to clinical reality.