Surface Proteins in the Differentiation of Stem Cells to Cardiomyocytes

心脏病是美国人的最大杀手。那些在心脏病发作中幸存下来的人患心脏病的风险更高 另一次发作，心力衰竭和中风。因此，需要开发修复方法 心脏和改善心脏病发作后的生活质量。再生医学的概念通过 干细胞作为一种恢复心脏功能的潜在疗法正受到人们的关注。跨栏 目前临床细胞疗法发展面临的问题是：(1)细胞的可获得性有限 用于选择纯细胞群体和跟踪分化的表面标记以及(2)缺乏 了解心肌细胞体外分化和体内发育的相互关系。从长远来看 该项目的目标是表征和了解干细胞分化。我们的目标是通过以下方式实现这一目标 识别和表征特定的细胞表面蛋白“条形码”有助于标记特定的细胞类型。 主要目标是首先解决对更多的细胞表面标记的需求，这些标记对 描述特定的细胞类型、阶段和谱系，特别是，我们将重点关注多能细胞， 心脏祖细胞和心室肌细胞。我们将使用最先进的分析生物化学和 在模型细胞系中鉴定和定量新表面蛋白的质谱学方法和这些 然后将评估蛋白质在活细胞分选中的效用，以获得纯的、确定的群体。 随后，我们将应用从体外工作中获得的知识来了解心肌细胞 动物的发育。具体地说，我们将确定体外标志物是否也存在于 胚胎发育过程中适宜的时间和空间。具体目标是为了开发 了解细胞表面标志物的功能以及分子事件在体外是否发生 分化类似于心肌细胞在体内发育过程中的分化。我所在的支持性环境 新的机构将通过提供访问最先进技术的途径来促进这项提议的继续成功 仪器，具有专业知识的导师，以及支持临床讨论的论坛 相关研究。

Heart disease is the single largest killer of Americans. Those that survive a heart attack have a higher risk of another attack, heart failure, and stroke. Consequently, there is a need to develop approaches for repairing the heart and improving the quality of life after a heart attack. The concept of regenerative medicine through the use of stem cells is gaining attention as a potential therapy for restoring cardiac function. Hurdles currently faced in the development of cellular therapies for the clinic are (1) the limited availability of cell surface markers for selecting pure cell populations and for tracking differentiation and (2) the lack of knowledge of how in vitro differentiation and in vivo development of cardiomyocytes correlate. The long term goal of this project is to characterize and understand stem cell differentiation. We aim to achieve this by identifying and characterizing specific cell surface protein 'barcodes' useful for marking specific cell types. The major objectives are to first address the need for more cell surface markers that are useful for characterizing specific cell types, stages, and lineages, and specifically, we will focus on pluripotent cells, cardiac progenitors, and ventricular cardiomyocytes. We will use state-of-the-art analytical biochemistry and mass spectrometry approaches to identify and quantify novel surface proteins in a model cell line and these proteins will then be evaluated for their utility in live cell sorting to obtain pure, defined populations. Subsequently, we will apply the knowledge gained from the in vitro work to understanding cardiac cell development in the animal. Specifically, we will determine whether the in vitro markers are also present at the appropriate time and space during embryonic development. The specific aims are designed to develop an understanding of the function of the cell surface markers and whether the molecular events during in vitro differentiation mimic those during in vivo development of cardiomyocytes. The supportive environment at my new institution will facilitate the continued success of this proposal by providing access to state-of-the art instrumentation, mentors with specialized expertise, and the forum to support discussion of clinically relevant research.