Surface Proteins in the Differentiation of Stem Cells to Cardiomyocytes

心脏病是美国人最大的单一杀手。那些心脏病发作后幸存下来的人， 再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， 干细胞作为一种恢复心脏功能的潜在疗法的应用正受到关注。障碍 目前在临床细胞疗法的开发中面临的问题是（1）细胞的有限可用性 用于选择纯细胞群体和用于跟踪分化的表面标记物，以及（2）缺乏 了解心肌细胞的体外分化和体内发育如何相互关联。长期 这个项目的目标是表征和理解干细胞分化。我们的目标是实现这一目标， 鉴定和表征可用于标记特定细胞类型的特定细胞表面蛋白“条形码”。 主要目标是首先解决对更多细胞表面标记物的需要，所述细胞表面标记物可用于 表征特定的细胞类型，阶段和谱系，具体来说，我们将专注于多能细胞， 心脏祖细胞和心室心肌细胞。我们将使用最先进的分析生物化学， 质谱方法来鉴定和定量模型细胞系中的新型表面蛋白， 然后评价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 动物的发展。具体来说，我们将确定体外标记物是否也存在于 在胚胎发育的适当时间和空间。具体目标是发展 了解细胞表面标志物的功能，以及体外培养过程中的分子事件是否 分化模拟心肌细胞体内发育期间的那些。在我的支持环境 一个新的机构将通过提供最先进的技术来促进这一提议的持续成功 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.