Ontogeny of erythroid lineage differentiation

红系谱系分化的个体发育

• 批准号: 7096319
• 负责人: James Palis
• 金额: $ 47.18万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7096319
• 关键词: cell differentiation; developmental genetics; embryo /fetus tissue /cell culture; embryonic stem cell; erythroid stem cell; erythropoiesis; gene expression profiling; gene targeting; genetic promoter element; in situ hybridization; laboratory mouse; mammalian embryology; microarray technology; molecular biology information system; small interfering RNA; transcription factor

DESCRIPTION (provided by applicant): The overall aim of my laboratory is to better understand the cellular and molecular underpinnings of the ontogeny of the mammalian hematopoietic system. We have recently found that primitive red cells in the embryo share significant similarities with their fetal and adult definitive counterparts, including an origin from unipotential progenitors, progressive maturational stages of nucleated erythroblasts, and terminal enucleation to become erythrocytes. However, primitive and definitive red cells also have significant differences in cell size, in transcriptional regulation, and in globin gene expression. We hypothesize that there are both shared and distinct stage-specific gene expression programs in differentiating primitive vs. definitive erythroid cells. In the first aim of this proposal, we will delineate a gene anatomy profile of primary primitive and definitive red blood cells at progressive stages of maturation. This data will be housed in a public database with a dedicated website for universal access. In the second aim, we will identify erythroid lineage-specific and stage-specific sets of genes and define erythroid-specific regulatory features using computationally assisted methods in collaboration with C. Stoeckert (U. Penn). The ability to compare similar stages of primitive red cells from the yolk sac and definitive red cells from the fetal liver and adult bone marrow gives us the unique opportunity to identify shared and differential programs of erythroid genes. In the third aim, we will test predictions generated in Aim 2 regarding the transcriptional regulation of lineage-specific and stage-specific gene cohorts. The function of specific candidate regulatory factors in erythroid differentiation will be screened by lentiviral siRNA-mediated gene knock-down in ES cell-derived primitive red cells. These studies will provide the research community with a comprehensive assembly of genes expressed at progressive stages of embryonic, fetal, and adult erythroid differentiation that will facilitate future characterization of structure-function relationships between expressed genes and red cell morphology, physiology, and disease phenotypes. Relevance of this research to public health. This research proposal is in response to a NIH program announcement (PA-03-150) to characterize all the genes in red blood cells. These studies will improve our understanding of the causes of anemia and will ultimately lead to curative treatments for children and adults with sickle cell disease and thalassemia syndromes.

描述（由申请人提供）：我实验室的总体目标是更好地了解哺乳动物造血系统个体发育的细胞和分子基础。我们最近发现胚胎中的原始红细胞与胎儿和成人的最终红细胞具有显着的相似性，包括起源于单能祖细胞、有核成红细胞的渐进成熟阶段以及最终去核成为红细胞。然而，原始红细胞和定形红细胞在细胞大小、转录调控和珠蛋白基因表达方面也存在显着差异。我们假设在区分原始红系细胞和定形红系细胞时存在共同的和不同的阶段特异性基因表达程序。在本提案的第一个目标中，我们将描绘成熟阶段的初级原始红细胞和最终红细胞的基因解剖学概况。这些数据将存储在公共数据库中，并设有专门的网站供普遍访问。在第二个目标中，我们将与 C. Stoeckert (U. Penn) 合作，使用计算辅助方法识别红细胞谱系特异性和阶段特异性基因组，并定义红细胞特异性调控特征。比较来自卵黄囊的原始红细胞和来自胎儿肝脏和成人骨髓的最终红细胞的相似阶段的能力为我们提供了识别红细胞基因的共享和差异程序的独特机会。在第三个目标中，我们将测试目标 2 中生成的有关谱系特异性和阶段特异性基因组转录调控的预测。将通过慢病毒 siRNA 介导的 ES 细胞来源的原始红细胞中的基因敲低来筛选特定候选调节因子在红系分化中的功能。这些研究将为研究界提供在胚胎、胎儿和成人红细胞分化进展阶段表达的基因的全面组装，这将有助于未来表征表达基因与红细胞形态、生理学和疾病表型之间的结构功能关系。这项研究与公共卫生的相关性。该研究提案是为了响应 NIH 计划公告 (PA-03-150)，以表征红细胞中的所有基因。这些研究将增进我们对贫血原因的了解，并最终为患有镰状细胞病和地中海贫血综合征的儿童和成人提供治疗方法。