Matrix-mediated endothelial differentiation of induced pluripotent stem cells

项目总结/摘要 美国有超过800万人患有外周动脉疾病（PAD）。PAD的一个特点是 血管内皮是一层内皮细胞（EC），对血管内皮细胞的功能障碍或损伤发挥控制作用。 血管反应性、重塑和血管生成。基于细胞的方法来恢复或再生 血管内皮细胞，从而增强血管生成反应缺血持有希望的PAD的治疗。一 EC的候选来源是诱导多能干细胞（iPSC），其来源于重编程的 体细胞iPSC保持无限的自我更新和分化为心血管系统的能力。 血统，包括EC。为了在治疗上利用iPSC，细胞必须首先分化成 然后有效地递送至缺血性疾病的部位。干细胞表型和 功能受到微环境因素的影响，包括细胞外基质（ECM），一种生物活性物质， 提供结构支持并调节细胞功能和表型的支架材料。ECM 细胞行为的调节是由整合素跨膜受体介导的，整合素跨膜受体将ECM连接到 细胞内细胞骨架并激活下游信号传导途径。ECM已被证明可以增强 多能干细胞的EC谱系的产量，但这些ECM是否是EC分化的最佳选择， 未知，因为没有系统的研究来评估基质介导的分化的作用。 该项目的目标是确定ECM在iPSC分化为EC中的作用， EC表型的维持和PAD动物模型中血管生成的治疗性增强。这 该项目将利用高通量ECM微阵列平台来优化基质介导的效率， iPSC分化为EC。EC分化过程中ECM-整合素相互作用的机制作用和 此外，亦会研究维修保养的问题。最后，将在动物模型中评估iPSC衍生的EC和ECM 用于血管再生。通过对ECM介导的细胞凋亡机制的深入了解， 分化和血管生成功能，申请人打算提供更强的知识基础 以及iPSC衍生的EC用于血管修复的临床开发和应用的改进方法。 申请人寻求建立一个终身制的学术生涯，以推进血管性心脏病的治疗。 利用生物工程和分子细胞生物学技术治疗疾病。申请人是博士后研究员， 她在斯坦福大学医学院接受干细胞和分子细胞方面的培训 John Cooke博士研究小组的技术，他是内皮细胞领域的一位知名研究者， 生物学和PAD疗法。将为她的培训和向独立过渡提供更多指导 由干细胞开发、基质生物学、组织工程、生物材料、 和生物统计学。

Project Summary/Abstract Over 8 million people in the US suffer from peripheral arterial disease (PAD). A feature of PAD is dysfunction or damage to the vascular endothelium, a layer of endothelial cells (ECs) that exerts control over vascular reactivity, remodeling and angiogenesis. Cell-based approaches to restore or regenerate the endothelium so as to enhance the angiogenic response to ischemia hold promise for the treatment of PAD. A candidate source of ECs is induced pluripotent stem cells (iPSCs), which are derived from reprogrammed somatic cells. The iPSCs maintain unlimited self renewal and the ability to differentiate into cardiovascular lineages, including ECs. In order to utilize iPSCs therapeutically, the cells must first be differentiated into the lineage of interest and then delivered efficiently to the site of ischemic disease. Stem cell phenotype and function are influenced by microenvironmental cues including the extracellular matrix (ECM), a biological scaffolding material that provides structural support and modulates cellular function and phenotype. ECM regulation of cell behavior is mediated by integrin transmembrane receptors that connect the ECM to the intracellular cytoskeleton and activate downstream signaling pathways. ECMs have been shown to enhance the yields of EC lineages of pluripotent stem cells, but whether these ECMs are optimal for EC differentiation is unknown because there has been no systematic study to assess the role of matrix-mediated differentiation. The goal of this project is to define the role of ECMs in the differentiation of iPSCs into ECs, maintenance of EC phenotype, and therapeutic enhancement of angiogenesis in animal models of PAD. This project will utilize a high-throughput ECM microarray platform to optimize the efficiency of matrix-mediated iPSC differentiation into ECs. The mechanistic role of ECM-integrin interactions during EC differentiation and maintenance will also be examined. Finally, iPSC-derived ECs and ECMs will be assessed in animal models of PAD for vascular regeneration. By gaining fundamental insights into mechanisms of ECM-mediated differentiation and angiogenic function, the applicant intends to provide a stronger foundation of knowledge and improved methods for the clinical development and application of iPSC-derived ECs for vascular repair. The applicant seeks to establish a tenure-track academic career in advancing the treatment of vascular diseases using bioengineering and molecular cell biology techniques. The applicant is a postdoctoral fellow in the Stanford University School of Medicine, where she is being trained in stem cell and molecular cellular techniques in the research group of Dr. John Cooke, a well-established investigator in the field of endothelial biology and PAD therapies. Additional guidance in her training and transition to independence will be provided by renowned experts in the fields of stem cell development, matrix biology, tissue engineering, biomaterials, and biostatistics.