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

描述(由申请人提供)： 斯坦福-约翰霍普金斯大学研究中心旨在更深入地了解分子途径，以提高核重新编程的效率，确保诱导多潜能细胞(IPSCs)的功能和安全性，为造血和内皮细胞的分化和纯化提供可靠的方案，并指导安全性和有效性的临床前研究的开创性工作。霍普金斯大学的研究小组提出了三个研究项目。项目1(S.Baylin)建议表征在重编程过程中发生的由DNMT介导的表观遗传事件，以适应干细胞的命运，并操纵这些事件以加强重编程和避免转化。项目2(E.Zambitis)建议利用这一新的发现来描述人类血管母细胞的特性，利用这种双能前体表达ACE，操纵ACE信号来促进成人HSC的出现，利用新的骨髓基质信号来促进成人HSC的出现，并确定被鉴定为血管母细胞或HSC特异性的新型miRNAs在指导HSC规范中的作用。该项目还将结合通过我们的中心和联盟获得的知识，以最佳地生成和临床前评估人类IPSO和成人HSC。项目3(A.Friedman)将确定允许Runx1指定HSC的机制，Runx1是成人HSC从血源性内皮中出现的主要转录调节因子。将评估Runxl异构体、Runxl磷酸化、Runxl与HDAC或ETS因子的相互作用以及Runx1与Notch、Wnt或BMP信号的协同作用。将使用全球RNA表达和芯片或ChlP-Seq方法识别相关的Runx1遗传靶点，表达分析将寻找在HSC中表达而不是在血管母细胞中表达的新调控因子。与我们的斯坦福同事合作开展的核心活动将包括IPSC与癌细胞和hESC的表观遗传学比较(S.Baylin)，以及表观遗传学、RNA表达和芯片数据的生物信息学分析(L.Cope)。我们预计，这些努力将导致对发育生物学的基本见解，以及血液病和血管疾病的新的翻译应用。

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.