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：
8759599
负责人：
STELLA T CHOU
金额：
$ 36.54万
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：
2014
资助国家：
美国
起止时间：
2014-09-01 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8759599
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 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 public health relevance 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的发展并引起人类疾病是生物医学中的一个基本问题。通过研究唐氏综合症（DS，三体术21，T21）和造血转录因子GATA-1，我们吸引了这个难题。 DS引起多种造血异常，包括多性炎，血小板减少症和两种相关的克隆疾病：瞬时骨髓增生性疾病（TMD）和急性巨毛质细胞性白血病（AMKL）。后一种疾病是多步进进展的一部分，需要体细胞GATA1突变，导致83个氨基酸截断的蛋白质称为GATA-1S。 GATA1中的类似种系突变会导致多倍体患者贫血。这些临床观察结果提出了几个有趣的问题：1）21染色体（HSA21）上的哪些基因调节造血？ 2）GATA-1氨基末端如何促进造血分化？ 3）在患有和没有T21的患者中，相同的GATA1突变如何引起不同的疾病？ 4）T21和GATA1突变如何唯一地协同引起骨髓增生？鼠模型提供了重要的信息，但并未完全概括人类疾病。我们正在使用人类胎儿肝标本和诱导多能干细胞（IPSC）研究这些问题，该干细胞（IPSC）是由DS，TMD和GATA1S相关性贫血的患者产生的。初步研究表明，具有T21和GATA-1S的IPSC表现出明显的造血异常，这些异常概括了相关人类疾病的许多方面。现在，我们将在具有T21，GATA-1S或两者兼有的IPSC中进行原始（蛋黄囊型）和确定性（胎儿肝型）造血的系统表征。我们将操纵IPSC中候选HSA21基因的表达，以识别负责DS相关血液异常的人。同时，我们将使用患者来源的IPSC和类似MEP的细胞系来学习 GATA1S突变如何通过识别与GATA-1氨基末端相互作用的蛋白质的努力来研究造血基因表达并研究相关机制。我们的研究将阐明GATA1和HSA21基因如何分别和一起调节造血。更普遍地，我们希望创建新的范式，在这种范式中，可以通过创建和操纵患者衍生的IPSC来建模其他造血疾病。