Matrix-mediated endothelial differentiation of induced pluripotent stem cells

基质介导的诱导多能干细胞的内皮分化

• 批准号: 8626434
• 负责人: Ngan F. Huang
• 金额: $ 24.4万
• 依托单位: PALO ALTO VETERANS INSTIT FOR RESEARCH
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2016-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8626434
• 关键词: Animal Model; Arterial Occlusive Diseases; Biocompatible Materials; Biological; Biological Process; Biology; Biomedical Engineering; Biometry; Blood Vessels; Blood capillaries; Cardiovascular system; Cell Differentiation process; Cell Lineage; Cell Maintenance; Cell Survival; Cell Therapy; Cell physiology; Cells; Cellular biology; Clinical; Cues; Cytoskeleton; Development; Disease; Embryonic Development; Endothelial Cells; Endothelium; Extracellular Matrix; Foundations; Functional disorder; Goals; Hindlimb; Histologic; Human; Hypoxia; In Vitro; Injury; Integrins; Ischemia; Isolated limb perfusion; Knowledge; Lasers; Lead; Maintenance; Mediating; Methods; Modeling; Molecular; Mus; Natural regeneration; Peripheral arterial disease; Phenotype; Pluripotent Stem Cells; Postdoctoral Fellow; Process; Public Health; Regulation; Research; Research Personnel; Role; Secondary to; Serum; Signal Pathway; Signal Transduction; Site; Somatic Cell; Source; Spectrum Analysis; Staging; Stem Cell Development; Stem cells; Supporting Cell; Techniques; Teratoma; Therapeutic; Time; Tissue Engineering; Tissues; Training; Universities; Vascular Diseases; Vascular Endothelial Cell; Vascular Endothelium; abstracting; angiogenesis; base; bioluminescence imaging; blood perfusion; capillary; career; cell behavior; density; functional improvement; improved; induced pluripotent stem cell; insight; interest; medical schools; nutrition; progenitor; receptor; repaired; response; scaffold; self-renewal; stem cell differentiation; stem cell therapy; success

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.

项目摘要/摘要 在美国，超过800万人患有外周动脉疾病(PAD)。PAD的一个特点是 血管内皮细胞功能障碍或损伤，血管内皮细胞(ECs)对 血管反应性、重塑和血管生成。基于细胞的方法来恢复或重新生成 血管内皮细胞从而增强对缺血的血管生成反应有望成为PAD的治疗手段。一个 内皮细胞的候选来源是诱导多能干细胞(IPSCs)，它来自于重新编程的来源 体细胞。IPSCs保持无限的自我更新和分化为心血管疾病的能力 血统，包括ECs。为了将IPSCs用于治疗，必须首先将细胞分化为 感兴趣的血统，然后有效地输送到缺血性疾病的部位。干细胞表型和 功能受到微环境信号的影响，包括细胞外基质(ECM)，一种生物 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连接细胞外基质和细胞外基质的整合素跨膜受体介导的 细胞内细胞骨架，并激活下游信号通路。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是否对EC分化是最佳的 未知，因为目前还没有系统的研究来评估基质介导的分化的作用。 本项目的目标是确定ECM在IPSC分化为ECS过程中的作用， 在PAD动物模型中维持EC表型，并在治疗上增强血管生成。这 项目将利用高通量ECM微阵列平台来优化基质中介的效率 IPSC向内皮细胞分化。细胞外基质-整合素相互作用在EC分化和分化中的机制作用 还将检查维护情况。最后，IPSC来源的ECs和ECM将在动物模型中进行评估 用于血管再生的垫片。通过对ECM介导的机制有基本的了解 分化与血管生成功能，申请者拟提供较强的知识基础 并改进了IPSC来源的内皮细胞用于血管修复的临床开发和应用方法。 申请者寻求在促进血管治疗方面建立终身教职的学术生涯。 使用生物工程和分子细胞生物学技术的疾病。申请人是一名博士后研究员。 斯坦福大学医学院，她正在那里接受干细胞和分子细胞方面的培训 约翰·库克博士的研究小组中的技术，他是内皮领域的知名研究员 生物学和PAD疗法。将为她的训练和向独立的过渡提供额外的指导 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。