GENETICS OF MYELODYSPLASTIC SYNDROMES

骨髓增生异常综合征的遗传学

基本信息

批准号：
8841404
负责人：
Matthew J Walter
金额：
$ 37.43万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-01 至 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8841404
关键词：
Acute Myelocytic Leukemia Age Apoptosis Biological Assay Blood Bone Marrow Cells Bone Marrow Transplantation CD34 gene Candidate Disease Gene Cell Cycle Cell Cycle Kinetics Cells Chromosome abnormality Chromosomes Chromosomes, Human, Pair 5 Complication Data Defect Development Disease Distal Dominant Genes Dose Dysmyelopoietic Syndromes Engineering Erythroid Progenitor Cells Event Gene Mutation Gene Targeting Genes Genetic Goals Hematopoiesis Hematopoietic Human Individual Ineffective Hematopoiesis Insertional Mutagenesis Knock-out Knockout Mice Laboratories Macrocytic Anemia Measures Modeling Molecular Chaperones Moloney Leukemia Virus Monitor Mus Mutate Newborn Infant Outcome Patients Penetrance Phenotype Population Prevalence Proteins RNA Interference Recurrence Reporter Retroviridae Risk S Phase Site Stem cells Syndrome Testing Viral Wild Type Mouse Work arm chromosome 5q loss defined contribution effective therapy erythroid differentiation improved integration site interstitial leukemia mouse development mouse model outcome forecast overexpression progenitor stem

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of this project is to identify and characterize genes that contribute to the development and progression of myelodysplastic syndromes (MDS). A better understanding of these genes may allow us to create effective therapies for patients with MDS and improve outcomes. An interstitial deletion involving one copy of the long arm of chromosome 5 (5q) is among the most common cytogenetic abnormalities in de novo and therapy-related MDS (25-40% of patients). We identified HSPA9 as a candidate gene located on the chromosome 5q31.2 commonly deleted region that contributes to altered hematopoiesis. We propose to define the mechanism of altered hematopoiesis caused by reduced levels of HSPA9 and to test whether Hspa9 haploinsufficiency contributes to MDS initiation and progression using primary human hematopoietic cells and heterozygous Hspa9 knock-out mice (Hspa9+/-), respectively, in the following specific aims: Specific Aim 1: We will determine whether the accelerated apoptosis and altered cell cycle kinetics observed following HSPA9 knockdown are p53-dependent. Because HSPA9 is known to physically interact with p53 in several models, we will determine whether they interact in primary human CD34+ cells. We will measure p53 activation in CD34+ cells following HSPA9 knockdown using a reporter assay and identify candidate p53-target genes responsible for the phenotype. We will validate the importance of p53 and the candidate genes using murine knockout models and RNA interference in primary human CD34+ cells. Specific Aim 2: We will establish whether haploinsufficiency of Hspa9 cooperates with Rps14 haploinsufficiency in mice. Mice lacking one copy of Rps14, a del(5q) gene, develop a macrocytic anemia and reduced progenitor cells. Most chromosome 5q deletions produce haploinsufficiency of both Hspa9 and Rps14 genes, suggesting that loss of both genes may cooperate in MDS. To directly test whether haploinsufficiency of Hspa9 and Rps14 cooperate during MDS initiation, we will intercross Hspa9+/- mice with mice lacking one copy of Rps14 and comprehensively characterize the hematopoiesis of singly or doubly heterozygous mice. Specific Aim 3: We will determine whether Hspa9 haploinsufficiency predisposes mice to develop leukemia. A devastating complication of MDS is the development of leukemia, which is highly-associated with deletions spanning chromosome 5q31.2. Identifying genes that contribute to del(5q31.2)-associated leukemia is vital to our understanding of this disease. To directly test whether haploinsufficiency of Hspa9 contributes to leukemia development, we will infect Hspa9+/- or wild-type littermate mice with a modified Moloney murine leukemia virus (MOL4070LTR retrovirus, a well established AML model) and monitor mice for the development of AML. We will identify and study candidate cooperating genes using mouse models.

描述（由申请人提供）：该项目的长期目标是识别和表征有助于骨髓增生综合征（MDS）发展和发展的基因。更好地了解这些基因可能使我们能够为MDS患者创建有效的疗法并改善预后。涉及染色体5（5q）长臂副本的间质缺失是从头和治疗相关的MDS中最常见的细胞遗传学异常之一（占患者的25-40％）。我们将HSPA9确定为位于5q31.2染色体上的候选基因，通常删除的区域有助于改变造血。我们建议定义由HSPA9水平降低引起的改变造血性的机制，并测试HSPA9单倍不足的不足效率是否有助于使用原发性人类造血细胞和杂合HSPA9敲出小鼠使用原发性造血细胞和杂合HSPA9敲出小鼠（HSPA9 +/-）（hspa9 +/-）（特定于特定的APS：aps：apts and aps：aps：aps：aps and ap and and aps：aps：aps：ap and and ap and aps：aps：aps：ap and and ap and aps：aps：aps：apts inters：aps：aps and inters 1： HSPA9敲低后观察到的循环动力学是p53依赖性的。由于已知HSPA9在几种模型中与p53物理相互作用，因此我们将确定它们是否在原代人CD34+细胞中相互作用。我们将使用记者测定法测量HSPA9敲低后CD34+细胞中的p53激活，并确定负责表型的候选p53目标基因。我们将使用鼠敲除模型和原代人CD34+细胞中的RNA干扰来验证p53和候选基因的重要性。具体目标2：我们将确定HSPA9的单倍弥补是否与小鼠的RPS14单倍弥补合作。缺少一份RPS14（5Q）基因的RPS14副本的小鼠会发展出大细胞性贫血和减少的祖细胞。大多数5Q染色体缺失可产生HSPA9和RPS14基因的单倍不足，这表明两个基因的损失可能在MDS中合作。为了直接测试HSPA9和RPS14在MDS启动过程中是否合作的单倍不足，我们将与缺少一份RPS14副本的小鼠间ross互变，并全面地表征单一或双重杂合小鼠的造血。具体目标3：我们将确定HSPA9单倍弥弥漫是否使小鼠患白血病。 MDS的破坏性并发症是白血病的发展，白血病与跨越5q31.2染色体的缺失高度相关。识别有助于DEL（5Q31.2）相关的白血病的基因对于我们对该疾病的理解至关重要。为了直接测试HSPA9的单倍不足是否有助于白血病的发育，我们将使用改良的Moloney鼠白血病病毒（MOL4070LTR逆转录病毒，良好的AML模型）感染HSPA9 +/-或Wild-type同变物小鼠，并监测AML开发的小鼠。我们将使用小鼠模型来识别和研究候选基因。