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

基本信息

批准号：
8607354
负责人：
Vijay Ganesh Sankaran
金额：
$ 26.31万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-03-01 至 2016-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8607354
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

项目摘要

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）：我们试图通过利用全基因组关联研究（GWAS）的强大统计发现来确定红细胞生产（红细胞生成）的新调节剂。尽管GWAS已经确定了与各种疾病或特征相关的人类基因组中成千上万的基因座，但这些基因座绝大多数的生物基础尚不清楚。这引起了人们对GWA对临床医学或功能生物学的更深入了解的价值的怀疑。我们以前已经利用了红细胞性格GWAS的结果来识别促红细胞生成的重要调节剂和球蛋白基因调节。这包括揭示胎儿到成年血红蛋白开关（BCL11A）的第一个特定调节剂的工作，以及揭示了Cyclin D3在末端红细胞生成中的关键作用的工作。在这个项目建议中，我们旨在扩展这些发现在人类中与红细胞性状有关的50多个基因座，试图揭示新的红血病调节剂。我们最初计划映射和识别基因座中的所有候选基因从红细胞性格GWAS揭示。这将通过利用识别常见人类变异的资源来实现，并允许我们推断链接不平衡模式。然后，我们计划使用原代人类红细胞细胞中的功能丧失综合发夹RNA（SHRNA）筛选在这些区域的功能上筛选所有候选基因。这将使我们能够从这些地区确定红细胞生成的候选监管者。然后，我们计划对人类细胞和小鼠模型中这些候选调节剂进行深入分析，以定义这些候选者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损失和功能扰动来实现。最后，我们旨在检查通过我们的研究鉴定的红细胞生成的调节剂是否可以操纵以改善当前体内红细胞产生的方法，这是直接临床相关性的问题。某些含抗原红细胞的血液供应的局限性构成了一个相当大的临床问题。在出现此类问题的情况下，这种离体衍生的红细胞可以替代标准输血。该项目有可能实质上推进生物医学研究的两个不同领域：（1）这些研究的发现将确定可能参与，修饰或帮助识别治疗各种形式的各种贫血的治疗途径的红细胞生成的新调节剂。我们先前在BCL11A上的工作说明了研究基本生物学问题（胎儿血红蛋白调节）如何导致有希望的治疗见解。（2）该项目中采用的功能方法可以提供资源，并作为未来研究的范式，旨在识别其他GWAS的生物基础。