Identifying Genome-wide Association Study-Nominated Regulators of Erythropoiesis

确定全基因组关联研究提名的红细胞生成调节因子

• 批准号: 9025971
• 负责人: Vijay Ganesh Sankaran
• 金额: $ 57.4万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9025971
• 关键词: Adult; Affect; Alleles; Anemia; Antigens; Applications Grants; Area; Biological; Biological Assay; Biology; Biomedical Research; Candidate Disease Gene; Cell Cycle Progression; Cell Proliferation; Cells; Clinical; Clinical Medicine; Cytokine Signaling; Disease; Erythrocytes; Erythroid; Erythroid Cells; Erythropoiesis; Fetal Hemoglobin; Gene Expression Regulation; Genes; Genetic Recombination; Globin; Grant; Hematopoiesis; Hemoglobin; Human; Human Genome; In Vitro; Individual; Knockout Mice; Lead; Linkage Disequilibrium; Maps; Medicine; Methods; Multipotent Stem Cells; Mus; Pattern; Pluripotent Stem Cells; Population; Process; Production; Red Blood Cell Count; Regulation; Resources; Role; Stem cells; Therapeutic; Transfusion; Variant; Vascular blood supply; Work; abstracting; clinically relevant; cyclin D3; erythroid differentiation; fetal; functional genomics; gain of function; genome wide association study; global health; improved; in vivo; insight; loss of function; mouse model; public health relevance; small hairpin RNA; success; therapy development; trait

DESCRIPTION (provided by applicant): We seek to identify new regulators of red blood cell production (erythropoiesis) by taking advantage of the robust statistical findings from genome-wide association studies (GWAS). While GWAS have identified thousands of loci across the human genome that are associated with various diseases or traits, the biological underpinnings for the vast majority of these loci remain unclear. This has given rise to skepticism about the value of GWAS for gaining a deeper understanding of clinical medicine or functional biology. We have previously utilized the results of erythroid trait GWAS to identify important regulators of erythropoiesis and globin gene regulation. This included work uncovering the first specific regulator of the fetal-to-adult hemoglobin switch, BCL11A, and work that revealed a critical role for cyclin D3 in terminal erythropoiesis. In this project proposal, we aim to extend these findings to over 50 loci associated with erythroid traits in humans in an attempt to uncover new regulators of erythropoiesis. We initially plan to map and identify all candidate genes in the loci revealed from erythroid trait GWAS. This will be accomplished by taking advantage of resources that identify common human variation and allow us to infer linkage disequilibrium patterns. We then plan to functionally screen all candidate genes in these regions using a loss-of-function pooled short hairpin RNA (shRNA) screen in primary human erythroid cells. This will allow us to identify candidate regulators of erythropoiesis from these regions. We then plan to perform in-depth analysis of these candidate regulators in human cells and in mouse models to define the exact role of these candidates in erythropoiesis. This will be accomplished through the use of both loss and gain-of-function perturbation of these candidate genes. Finally, we aim to examine whether the regulators of erythropoiesis identified through our studies can be manipulated to improve the current methods of ex vivo red blood cell production, which is a problem of immediate clinical relevance. Limitations in the blood supply of certain antigen-containing red blood cells pose a considerable clinical problem. Such ex vivo-derived red cells could serve as an alternative to standard transfusions in cases where such problems arise. This project has the potential to substantially advance two distinct areas of biomedical research: (1) The findings from these studies will identify new regulators of erythropoiesis that could be involved in, modifiers of, or help identify therapeutic avenues to treat various forms of anemia. Our prior work on BCL11A illustrates how studying a fundamental biological problem (fetal hemoglobin regulation) can result in promising therapeutic insight. (2) The functional approaches undertaken in this project could provide resources and serve as a paradigm for future research aimed at identifying the biological underpinnings of other GWAS.

描述(由申请人提供)：我们寻求通过利用全基因组关联研究(GWAS)的强大统计结果来确定红细胞生成(红细胞生成)的新调节因素。虽然Gwas已经在人类基因组中确定了数千个与各种疾病或特征相关的基因座，但这些基因座中绝大多数的生物学基础仍然不清楚。这引起了人们对GWAs对于更深入地理解临床医学或功能生物学的价值的怀疑。我们之前已经利用红系特征GWAs的结果来确定红系生成和珠蛋白基因调控的重要调节因子。这包括发现胎儿到成人血红蛋白开关的第一个特定调节因子BCL11A，以及揭示细胞周期蛋白D3在终末红细胞生成中的关键作用的工作。在这个项目提案中，我们的目标是扩展这些发现 50多个与人类红系特征相关的基因座，试图发现红细胞生成的新调节因素。我们最初的计划是绘制和识别这些基因座上的所有候选基因 从红系特征中揭示出来。这将通过利用资源来实现，这些资源可以识别共同的人类变异，并允许我们推断连锁不平衡模式。然后，我们计划在原代人类红系细胞中使用功能缺失的短发夹状RNA(ShRNA)筛选这些区域的所有候选基因。这将使我们能够从这些地区确定红细胞生成的候选调节者。然后，我们计划在人类细胞和小鼠模型中对这些候选调控因子进行深入分析，以确定这些候选调控因子在红细胞生成中的确切作用。这将通过使用这些候选基因的功能丢失和功能获得扰动来实现。最后，我们的目标是检查通过我们的研究确定的红细胞生成调节因子是否可以被操纵来改进目前的体外红细胞生成方法，这是一个直接与临床相关的问题。某些含有抗原的红细胞的血液供应受限是一个相当大的临床问题。在出现此类问题的情况下，这种来自体外的红细胞可以作为标准输血的替代方案。该项目有可能极大地推动生物医学研究的两个不同领域：(1)这些研究的结果将确定新的红细胞生成调节剂，这些调节剂可能涉及、修饰或帮助确定治疗各种形式贫血的途径。我们之前在BCL11A上的工作说明了研究一个基本的生物学问题(胎儿血红蛋白调节)是如何产生有希望的治疗见解的。(2)该项目采用的功能方法可提供资源，并可作为未来研究的范例，目的是确定其他全球气候变化系统的生物学基础。