GATA1 Mutation in Defective Erythropoiesis

红细胞生成缺陷中的 GATA1 突变

基本信息

批准号：
8651635
负责人：
John D Crispino
金额：
$ 33.04万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-18 至 2018-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8651635
关键词：
Acute Megakaryocytic Leukemias Affect Anemia Animals Binding Blood Platelets Cells ChIP-seq Chromatin Congenital Anemia Congenital Disorders Congenital dyserythropoietic anemia Coupled DNA DNA Binding Data Data Set Defect Development Diamond-Blackfan anemia Down Syndrome Embryo Erythroblasts Erythrocytes Erythroid Erythroid Cells Erythroid Progenitor Cells Erythropoiesis Failure Fetal Liver Fingers Friends GATA1 gene Gene Expression Gene Expression Regulation Gene Targeting Genes Grant Gray unit of radiation dose Hematological Disease Hematopoietic Hemoglobin Histones Human Knock-in Mouse Laboratories Lead Length Megakaryocytes Mus Mutation N-terminal Names Nuclear Patients Physical condensation Porphyrias Pregnancy Process Protein Isoforms Proteins Proteomics Research Site Syndrome Thrombocytopenia base chromatin immunoprecipitation cofactor dyserythropoietic anemia erythroid Kruppel-like factor genome-wide in vivo induced pluripotent stem cell insight interest mast cell mitochondrial autophagy mutant next generation sequencing novel novel strategies progenitor public health relevance research study transcription factor transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): GATA1 is a transcription factor that is required for survival and maturation of erythroid cells. In the absence of GATA1, mouse embryos die from anemia in mid-gestation. In humans, GATA1 mutations are associated with a spectrum of blood disorders, including congenital dyserythropoietic anemia and thrombocytopenia, porphyria, and Diamond-Blackfan Anemia (DBA). In the presence of trisomy 21, GATA1 mutations that delete the N- terminus lead to Down syndrome associated Acute Megakaryoblastic Leukemia (DS-AMKL). We recently demonstrated that a GATA1 mutant, which fails to bind FOG1 and is associated with rare cases of cases of dyserythropoietic anemia, fails to bind chromatin in the same manner as wild-type GATA1. This differential chromatin binding allows GATA1 to promote mast cell formation instead of red cell or megakaryocyte development. In a similar fashion, we have recently discovered that GATA1 molecules that lack the N-terminal activation domain, seen in both DBA and DS-AMKL, also fail to bind chromatin in the same way as full-length GATA1. Of interest, genes that are not properly bound or regulated by GATA1s include critical red cell genes such as Alas2, Slc4a1, and Klf1 (EKLF). In this grant, we will precisely define the requirement for the N- terminus of GATA1 in red cell development. Our aim is to discover how the GATA1 mutations that lead to expression of GATA1s in place of the full-length protein cause defects in the erythroid lineage and congenital anemia, including DBA. Our overarching hypothesis is that the reduced chromatin binding and aberrant gene regulation by GATA1s leads to impaired specification and terminal differentiation of red blood cells. Our aims are to: 1 Correlate chromatin occupancy of GATA1s with gene expression defects in primary GATA1s knock-in erythroid progenitor cells to identify key direct target genes that are dysregulated; 2) Investigate the consequences of the GATA1s mutation on erythroid specification and differentiation; and 3) Determine if loss of the N-terminus reduces the interaction with essential cofactors and in turn affects their chromatin occupancy. The research described in this proposal will greatly expand our insights into how loss of the N-terminus of GATA1 alters erythropoiesis and will benefit patients with congenital anemia and DS-AMKL.

描述（由申请人提供）：GATA1是红系细胞存活和成熟所需的转录因子。如果缺乏 GATA1，小鼠胚胎会在妊娠中期因贫血而死亡。在人类中，GATA1 突变与一系列血液疾病相关，包括先天性红细胞生成障碍性贫血和血小板减少症、卟啉症和 Diamond-Blackfan 贫血 (DBA)。在 21 三体存在的情况下，删除 N 末端的 GATA1 突变会导致唐氏综合症相关的急性巨核细胞白血病 (DS-AMKL)。我们最近证明，GATA1 突变体无法以与野生型 GATA1 相同的方式结合染色质，该突变体无法结合 FOG1 并且与罕见的红细胞生成障碍性贫血病例相关。这种差异化的染色质结合使 GATA1 能够促进肥大细胞的形成，而不是红细胞或巨核细胞的发育。以类似的方式，我们最近发现缺乏 N 端激活结构域的 GATA1 分子（在 DBA 和 DS-AMKL 中均可见）也无法以与全长 GATA1 相同的方式结合染色质。有趣的是，未正确结合或受 GATA1 调节的基因包括关键的红细胞基因，例如 Alas2、Slc4a1 和 Klf1 (EKLF)。在这笔资助中，我们将精确定义红细胞发育中 GATA1 N 末端的要求。我们的目标是发现导致 GATA1 表达代替全长蛋白的 GATA1 突变如何导致红系谱系缺陷和先天性贫血（包括 DBA）。我们的总体假设是，GATA1 染色质结合的减少和基因调控的异常会导致红细胞的规格和终末分化受损。我们的目标是： 1 将 GATA1 的染色质占据与原代 GATA1 敲入红系祖细胞中的基因表达缺陷相关联，以识别失调的关键直接靶基因； 2) 研究GATA1s突变对红系规范和分化的影响； 3) 确定 N 末端的丢失是否会减少与必需辅助因子的相互作用，进而影响其染色质占据。该提案中描述的研究将极大地扩展我们对 GATA1 N 末端缺失如何改变红细胞生成的认识，并将使先天性贫血和 DS-AMKL 患者受益。