Systematic Genetic Dissection of Human Erythropoiesis

项目概要/摘要 人类在红细胞生成（红细胞生成）过程中差异很大。这种变化 导致疾病，包括多种形式的贫血。这种变异大部分可归因于遗传 在人群中发现的多态性。然而，与大多数其他复杂疾病和特征一样， 变异对红细胞生成过程的影响仍然知之甚少。我们取得了巨大 在开始破译人类遗传变异如何影响前五年的红细胞生成方面取得了进展 这是NIDDK多年来的资助。我们率先使用大规模并行报告基因测定作为 一种系统地剖析非编码遗传变异如何从全基因组关联中揭示的方法 GWAS研究（GWAS）在功能上影响转录调控元件。我们还利用了CRISPR/Cas9 基因组编辑以支持内源性背景下的这些发现。在这次竞争性的续约申请中，我们 旨在振兴这些研究，并直接解决人类遗传变异如何塑造非编码基因， 红细胞生成的基因组景观。我们将在先前研究的基础上，采用创新的新方法 在原代人类红细胞中提出这些问题。我们最近进行了一次最大的GWAS 红细胞性状的研究，并确定了超过1,800个不同的相关基因座。我们建议利用一些 尖端的基因组工具，使我们能够从这些新兴的大规模遗传学中获得重要的见解， 问题研究首先，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 对连接的转录调控元件的影响。其次，我们将系统地鉴定靶基因 这可能会受到携带性状相关变异的调节元件的影响。最后，我们将解剖 这些变体的子集通过其影响靶基因并改变人红细胞生成的作用机制。 未来五年的研究不仅将大大促进我们对人类的理解， 红细胞生成和相关疾病，但这项工作也将作为一个范例，系统解剖 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.