Midwest Progenitor Cell Consortium

描述(由申请人提供)： 中西部先驱Ciell联盟是威斯康星大学麦迪逊分校和UMN之间的合作，将成为NHLBI先驱细胞生物学联盟的研究中心。其目标是开发新的策略来将人类多能干细胞和体细胞转化为造血干细胞(HSCs)，并通过鉴定导致前HSCs和心脏前体细胞形成的遗传程序以及激活谱系受限细胞中的自我更新程序来自我更新心脏前体细胞。项目1，“从人ES/iPS细胞和体细胞中新生成造血干细胞CEU”，目的是：利用靶向FOXF1、GATA-2、GATA-3、RUNX1和SCL基因的人ES细胞系，鉴定血管内皮细胞和造血祖细胞的中胚层祖细胞的层级；确定在小鼠和人类胚胎发育过程中导致造血细胞和HSCs形成的最关键的分子事件，以及人类多能干细胞的体外分化；开发从人ES/iPS细胞和体细胞中新生成HSCs的技术。项目2“来自人类ES/iPS细胞和体细胞的心血管祖细胞和心肌细胞”，目的是：分离和确定来源于多能干细胞的人类胚胎心血管祖细胞和心室肌细胞；以及使用转录因子对人类多能干细胞和体细胞进行重新编程，以生成心血管祖细胞和心室肌细胞。项目3“用于重编程/跨分化的人工转录因子”的目标是：设计在原代成纤维细胞中激活Oct4、Sox2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；以及结合ATF在原代成纤维细胞中诱导多能性。项目4“造血和心血管前体亲和力试剂的体外快速生成”，目标是：开发快速生成与高亲和力和特异性的跨膜蛋白靶标结合的适配子的方法；开发利用高通量DNA测序技术加速发现具有亚纳米分子亲和力的适配子的方法；以及为心血管和血液前体的表面标记生成高亲和力的适配子。

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.