Systematic Genetic Dissection of Human Erythropoiesis

项目摘要/摘要 人类在红细胞生成(红细胞生成)的过程中有很大的差异。这样的变异 导致疾病，包括多种形式的贫血。这种变异很大程度上可归因于基因 在种群中发现了多态。然而，就像大多数其他复杂的疾病和特征一样，这种情况 变异对红细胞生成过程的影响仍然知之甚少。我们已经取得了巨大的成就 开始破译人类基因变异如何影响前五年的红细胞生成的进展 多年来NIDDK提供的这笔赠款。我们率先使用大量平行的记者分析作为一种 系统剖析全基因组关联揭示非编码遗传变异的方法 研究(GWAS)从功能上影响转录调控元件。我们还利用了CRISPR/CAS9 基因组编辑在内生的背景下支持这些发现。在这项竞争性续订申请中，我们 目的是为这些研究注入活力，并直接解决人类基因变异如何塑造非编码 红细胞生成的基因组图谱。我们将在以前研究的基础上，使用创新的新方法 在原代人类红系细胞中提出这些问题。我们最近进行了一次规模最大的全球气候变化分析 并确定了1,800多个不同的相关基因座。我们建议利用一些 尖端基因组工具，使我们能够从这些新兴的大规模基因 学习。首先，我们的目标是定义这些关联背后的假定因果变量及其功能 对相互关联的转录调控元件的影响。第二，我们将系统地识别靶基因 这可能受到含有与性状相关的变异的调控元件的影响。最后，我们将剖析 这些变异的一个子集影响靶基因并改变人类红细胞生成的作用机制。 未来五年拟议的研究不仅将极大地促进我们对人类的了解 红细胞生成和相关疾病，但这项工作也将成为系统剖析 其他复杂的疾病和特征。

Project Summary/ Abstract Humans vary considerably in the process of red blood cell production (erythropoiesis). Such variation contributes to diseases, including numerous forms of anemia. Much of this variation is attributable to genetic polymorphisms found in the population. However, as with most other complex diseases and traits, how such variation impacts the process of erythropoiesis remains poorly understood. We have made tremendous progress in beginning to decipher how human genetic variation impacts erythropoiesis during the first five years of this grant from the NIDDK. We have pioneered the use of massively parallel reporter assays as an approach to systematically dissect how non-coding genetic variation revealed from genome-wide association studies (GWAS) functionally impacts transcriptional regulatory elements. We have also utilized CRISPR/Cas9 genome editing to bolster these findings in an endogenous context. In this competitive renewal application, we aim to invigorate these studies and directly address how human genetic variation shapes the non-coding genomic landscape of erythropoiesis. We will build upon our prior studies and use innovative new approaches to ask these questions in primary human erythroid cells. We have recently conducted one of the largest GWAS of erythroid traits and have identified over 1,800 distinct associated loci. We propose to utilize a number of cutting-edge genomic tools that will allow us to gain important insights from these emerging large-scale genetic studies. First, we aim to define putative causal variants underlying these associations and their functional effects on the linked transcriptional regulatory elements. Second, we will systematically identify target genes that may be affected by the regulatory elements harboring trait-associated variation. Finally, we will dissect mechanisms of action by which a subset of these variants impact target genes and alter human erythropoiesis. Not only will the proposed studies over the next five years significantly advance our understanding of human erythropoiesis and related diseases, but this work will also serve as a paradigm for the systematic dissection of other complex diseases and traits.