Identifying Genome-wide Association Study-Nominated Regulators of Erythropoiesis

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）的强大统计结果来鉴定红细胞生成（红细胞生成）的新调节剂。虽然GWAS已经在人类基因组中确定了数千个与各种疾病或性状相关的基因座，但这些基因座中绝大多数的生物学基础仍不清楚。这引起了对GWAS的价值的怀疑，以获得更深入的了解临床医学或功能生物学。我们以前利用红系性状GWAS的结果，以确定红细胞生成和珠蛋白基因调控的重要调节。这包括揭示胎儿到成人血红蛋白转换的第一个特定调节因子BCL 11A的工作，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。在本项目建议书中，我们旨在扩展这些发现 超过50个与人类红细胞性状相关的基因座，试图发现红细胞生成的新调节因子。我们最初计划定位和识别位点中的所有候选基因 从红细胞性状GWAS中发现。这将通过利用资源，确定共同的人类变异，使我们能够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丧失的合并短发夹RNA（shRNA）筛选功能筛选这些区域中的所有候选基因。这将使我们能够从这些区域识别红细胞生成的候选调节因子。然后，我们计划在人类细胞和小鼠模型中对这些候选调节剂进行深入分析，以确定这些候选物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丧失和功能获得扰动来实现。最后，我们的目的是检查是否通过我们的研究确定的红细胞生成的调节剂可以被操纵，以改善目前的方法，离体红细胞生产，这是一个直接的临床相关性的问题。某些含抗原的红细胞的血液供应的限制造成了相当大的临床问题。在出现这些问题的情况下，这种体外衍生的红细胞可以作为标准输血的替代品。该项目有可能实质性地推进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因子，这些调节因子可能参与、修饰或帮助确定治疗各种形式贫血的治疗途径。我们之前对BCL11A的研究说明了研究一个基本的生物学问题（胎儿血红蛋白调节）如何产生有希望的治疗见解。(2)在这个项目中进行的功能方法可以提供资源，并作为一个范例，为未来的研究，旨在确定其他GWAS的生物学基础。

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.