Modeling genetic modifiers of hematopoiesis with induced pluripotent stem cells

用诱导多能干细胞模拟造血的遗传修饰剂

基本信息

批准号：
9049947
负责人：
STELLA T CHOU
金额：
$ 2.24万
依托单位：
CHILDREN'S HOSP OF PHILADELPHIA
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9049947
关键词：
4 year old Acute Megakaryocytic Leukemias Address Amino Acids Anemia Binding Biological Biological Models Blast Cell Blood Blood Cells Blood typing procedure Cell Line Child Chromatin Chromosomes, Human, Pair 21 Clinical Collaborations Communities DNA Development Diamond-Blackfan anemia Disease Disease model Down Syndrome E2F1 gene Erythro Erythroid Erythropoiesis Exhibits Fetal Liver Functional disorder GATA1 gene Gene Expression Gene Expression Profile Gene Targeting Genes Genetic Genetic Models Genotype Germ-Line Mutation Goals Health Hematological Disease Hematopoiesis Hematopoietic Hereditary Disease Human Infant Inherited Macrocytic Anemia Megakaryocytes Megakaryocytopoieses Methods Modeling Molecular Abnormality Mus Mutation Myeloproliferation Myeloproliferative disease Neutropenia Patients Phenocopy Phenotype Polycythemia Production Property Proteins Protocols documentation Regulation Repression Retinoblastoma Protein Role Specimen Study models Syndrome Testing Thrombocytopenia Tissues Transcription factor genes Work Yolk Sac chromatin immunoprecipitation cofactor fetal histone modification human GATA1 protein human disease human subject improved induced pluripotent stem cell insight interest leukemia mouse model novel postnatal progenitor tool transcription factor transient myeloproliferative disorder

项目摘要

DESCRIPTION (provided by applicant): Understanding how genes interact to coordinate tissue development and cause human disease is a fundamental problem in biomedicine. We are drawn to this puzzle through studies of Down syndrome (DS, trisomy 21, T21) and the hematopoietic transcription factor GATA-1. DS causes multiple hematopoietic abnormalities including polycythemia, thrombocytopenia and two related clonal disorders: transient myeloproliferative disorder (TMD) and acute megakaryoblastic leukemia (AMKL). The latter disorders are part of a multi-step progression that requires somatic GATA1 mutations resulting in an 83 amino acid truncated protein termed GATA-1s. Similar germline mutations in GATA1 cause anemia in euploid patients. These clinical observations raise several interesting questions: 1) what genes on chromosome 21 (HSA21) regulate hematopoiesis? 2) How does the GATA-1 amino terminus facilitate hematopoietic differentiation? 3) How do the same GATA1 mutations cause different diseases in patients with and without T21? and 4) How do T21 and GATA1 mutations synergize uniquely to cause myeloproliferation? Murine models have provided important information, but do not fully recapitulate the human diseases. We are studying these problems using human fetal liver specimens and induced pluripotent stem cells (iPSCs) generated from patients with DS, TMD, and GATA1s-associated anemias. Preliminary studies indicate that iPSCs with T21 and GATA-1s exhibit distinct hematopoietic abnormalities that recapitulate many aspects of the associated human disorders. Now, we will perform systematic characterization of primitive (yolk sac-type) and definitive (fetal liver-type) hematopoiesis in iPSCs with T21, GATA-1s or both. We will manipulate the expression of candidate HSA21 genes in iPSCs to identify those responsible for DS-associated blood abnormalities. In parallel, we will use our patient-derived iPSCs and a MEP-like cell line to study how GATA1s mutations dysregulate hematopoietic gene expression and investigate the associated mechanisms through efforts to identify proteins that interact with the GATA-1 amino terminus. Our studies will elucidate how GATA1 and HSA21 genes, separately and together, modulate hematopoiesis. More generally, we hope to create new paradigms in which other hematopoietic diseases can be modeled through creation and manipulation of patient-derived iPSCs.

描述（由申请人提供）：了解基因如何相互作用以协调组织发育并导致人类疾病是生物医学中的一个基本问题。通过对唐氏综合症（DS、21 三体、T21）和造血转录因子 GATA-1 的研究，我们被这个难题所吸引。 DS 会导致多种造血异常，包括红细胞增多症、血小板减少症和两种相关的克隆性疾病：短暂性骨髓增殖性疾病 (TMD) 和急性巨核细胞白血病 (AMKL)。后一种疾病是多步骤进展的一部分，需要体细胞 GATA1 突变，产生 83 个氨基酸的截短蛋白，称为 GATA-1。 GATA1 中类似的种系突变会导致整倍体患者贫血。这些临床观察结果提出了几个有趣的问题：1）21 号染色体 (HSA21) 上的哪些基因调节造血作用？ 2）GATA-1氨基末端如何促进造血分化？ 3）相同的GATA1突变如何在有和没有T21的患者中引起不同的疾病？ 4) T21 和 GATA1 突变如何独特地协同作用导致骨髓增殖？小鼠模型提供了重要信息，但并未完全概括人类疾病。我们正在使用人类胎儿肝脏样本和来自 DS、TMD 和 GATA1 相关贫血患者的诱导多能干细胞 (iPSC) 研究这些问题。初步研究表明，具有 T21 和 GATA-1 的 iPSC 表现出明显的造血异常，概括了相关人类疾病的许多方面。现在，我们将使用 T21、GATA-1 或两者对 iPSC 中的原始（卵黄囊型）和决定性（胎肝型）造血进行系统表征。我们将操纵 iPSC 中候选 HSA21 基因的表达，以确定那些与 DS 相关的血液异常负责的基因。与此同时，我们将使用源自患者的 iPSC 和 MEP 样细胞系来研究 GATA1 突变如何失调造血基因表达，并通过努力鉴定与 GATA-1 氨基末端相互作用的蛋白质来研究相关机制。我们的研究将阐明 GATA1 和 HSA21 基因如何单独和共同调节造血作用。更一般地说，我们希望创建新的范例，通过创建和操作患者来源的 iPSC 来模拟其他造血系统疾病。