Midwest Progenitor Cell Consortium

描述（由申请人提供）： 中西部祖细胞Ciell联盟是威斯康星大学麦迪逊分校和UMN之间的合作，将作为NHLBI祖细胞生物学联盟内的研究中心。本研究的目标是通过鉴定导致前-HSC和心脏祖细胞形成的遗传程序和激活谱系限制细胞中的自我更新程序，开发将人类多能干细胞和体细胞转化为造血干细胞（HSC）和自我更新心脏祖细胞的新策略。项目1，“The De Novo Generation of Hematopoietic Stem CeUs from Human ES/iPS Cells and Somatic Cells”，目标是：使用具有靶向FOXF 1、加塔-2、加塔-3、RUNX 1和SCL基因的人ES细胞系鉴定血管内皮和造血谱系的中胚层祖细胞的层次;确定小鼠和人类胚胎发生过程中导致造血细胞和HSC形成的最关键分子事件，开发从人ES/iPS细胞和体细胞重新产生HSC的技术。项目2，“来自人ES/iPS细胞和体细胞的心血管祖细胞和心肌细胞”，目标是：分离和定义来自多能干细胞的人胚胎心血管祖细胞和心室肌细胞;并使用转录因子重编程人多能干细胞和体细胞以产生心血管祖细胞和心室肌细胞。项目3，“用于重编程/转分化的人工转录因子”，目标是：设计激活原代成纤维细胞中Oct 4、Sox 2或Nanog表达的ATF;进行全基因组定位分析、CSI和转录组分析以确定所设计的ATF的基因靶点和特异性;以及联合收割机ATF以诱导原代成纤维细胞中的多能性。项目4，“造血和心血管前体亲和试剂的体外快速生成”，目标是：开发快速生成以高亲和力和特异性结合跨膜蛋白靶点的适体的方法;开发使用高通量DNA测序技术加速发现具有亚纳摩尔亲和力的适体的方法;并产生用于心血管和血液前体的表面标志物的高亲和力适体。

DESCRIPTION (provided by applicant): The Midwest Progenitor Ciell Consortium is a collaboration between the UW-Madison and the UMN which will serve as a research hub within the NHLBI's Progenitor Cell Biology Consortium. The goal is to develop new strategies to convert human pluripotent stem and somatic cells into hematopoietic stem cells (HSCs) and selfrenewing cardiac progenitor cells through identification of genetic program leading to formation of pre-HSCs and cardiac progenitors and activation of self-renewal program in lineage-restricted cells. Project 1, "The De Novo Generation of Hematopoietic Stem CeUs from Human ES/iPS Cells and Somatic Cells," aims are: Identify the hierarchy of mesodermal progenitors of vascular endothelial and hematopoietic lineages using human ES cell lines with targeted FOXFl, GATA-2, GATA-3, RUNXl, and SCL genes; Determine the most critical molecular events leading to formation of hematopoietic cells and HSCs during embryogenesis in mouse aind human, and following in vitro differentiation of human pluripotent stem cells; Develop technologies for de novo generation of HSCs from human ES/iPS cells and somatic cells. Project 2, "Cardiovascular Progenitors and Cardiomyocytes from Human ES/iPS Cells and Somatic Cells," aims are: Isolate and define human embryonic cardiovascular progenitors and ventricular myocytes derived from pluripotent stem cells; and reprogram human pluripotent stem cells and somatic cells using transcription factors to generate cardiovascular progenitors and ventricular myocytes. Project 3, "Artificial Transcription Factors for Reprogramming/Transdifferentation," aims are: Design ATFs that activate expression of Oct4, Sox2 or Nanog in primary fibroblasts; Perform genome-wide location analysis, CSI and transcriptome analysis to determine gene targets and specificity ofthe designed ATFs; and combine ATFs to induce pluripotency in primary fibroblasts. Project 4, "Rapid In Vitro Generation of Affinity Reagents for Hematopoietic and Cardiovascular Precursors," aims are: Develop methods for rapidly generating aptamers that bind to transmembrane protein targets with high affinity and specificity; Develop methods to accelerate discovery of aptamers with sub nanomolar affinities using high-throughput DNA sequencing technologies; and generate high affinity aptamers for surface markers of cardiovascular and blood precursors.