Basic and Translational Research of iPSC-Based hematologic and Vascular Therapies

描述（由申请人提供）： 斯坦福-约翰霍普金斯研究中心旨在更深入地了解分子途径，以提高核重编程的效率，确保诱导多能细胞（iPSCs）的功能和安全性，为造血和内皮谱系的分化和纯化提供强大的方案，并指导安全性和有效性的临床前研究的开创性工作。霍普金斯小组提出了三个研究项目。项目1（S. Baylin）提出表征在干细胞命运的重编程期间发生的由DNMT介导的表观遗传事件，并操纵这些事件以增强重编程并避免转化。项目2（E. Zambidis）提出利用这种双能祖细胞表达ACE的新发现来表征人成血管细胞，操纵ACE信号以促进成体HSC的出现，利用新的骨髓基质信号以促进成体HSC的出现，并确定被鉴定为成血管细胞或HSC特异性的新miRNA在指导HSC特化中的作用。该项目还将结合联合收割机在我们的中心和联盟中获得的知识，以最佳方式生成和临床前评估人类IPSO和成人HSC。项目3（A. Friedman）将鉴定允许通过Runxl（成体HSC从生血内皮出现的主要转录调节因子）特异化HSC的机制。将评估Runxl同种型、Runxl磷酸化、Runxl与HDAC或Ets因子的相互作用以及Runxl与Notch、Wnt或BMP信号传导的合作的作用。将进行使用全局RNA表达和ChIP-芯片或ChIP-Seq方法鉴定相关Runxl遗传靶标，并且表达分析将寻找在HSC中而不是在成血管细胞中表达的新型调节剂。与我们的斯坦福大学同事合作进行的CORE活动将包括IPSC与癌细胞和hESC的表观遗传学比较（S. Baylin）和表观遗传学、RNA表达和ChIP数据的生物信息学分析（L.科普）。我们预计，这些努力将导致在发育生物学的基本见解和新的，血液和血管疾病的翻译应用。

DESCRIPTION (provided by applicant): The Stanford-Johns Hopkins Research Hub intends to gain a deeper understanding of molecular pathways to enhance the efficiency of nuclear reprogramming, to ensure the function and safety of induced pluripotentlal cells (iPSCs), to provide robust protocols for differentiation and purification of hematopoietic and endothelial lineages, and to guide pioneering work in pre-clinical studies of safety and efficacy. The Hopkins group proposes three research projects. PROJECT 1 (S. Baylin) proposes to characterize epigenetic events mediated by DNMT occuring during reprogramming to the stem cell fate and to manipulate these events to enhance reprogramming and avoid transformation. PROJECT 2 (E. Zambidis) proposes to characterize the human hemangioblast, taking advantage of the novel finding that this bipotent progenitor is expresses ACE, to manipulate ACE signaling to favor emergence of adult HSC, to utilize novel marrow stromal signals to favor emergence of adult HSC, and to determine the role of novel miRNAs identified as hemangioblast or HSC-specific in directing HSC specification. This project will also combine knowledge gained throughout our Hub and the Consortium to optimally generate and pre-clinically evaluate human IPSO and adult HSC. PROJECT 3 (A. Friedman) will identify the mechanisms allowing HSC specification by Runxl, a master transcriptional regulator of adult HSC emergence from hemogenic endothelium. The role of Runxl isoforms, Runxl phopshorylation, Runxl interaction with HDACs or Ets factors, and Runxl cooperation with Notch, Wnt, or BMP signaling will be evaluated. Identification of relevant Runxl genetic targets using global RNA expression and ChiP-chip or ChlP-Seq approaches will be undertaken, and expression analyses will seek novel regulators, expressed in HSC but not the hemangioblast. CORE activities conducted in collaboration with our Stanford colleagues will include epigenetic comparison of IPSC to cancer cells and to hESC (S. Baylin) and bioinformatics analysis of epigenetic, RNA expression, and ChIP data (L. Cope). We anticipate that these efforts will lead to basic insights in developmental biology and to novel, translational applications for hematologic and vascular disorders.