Genetic Identification of Novel Genes Critical in Erythropoiesis

红细胞生成关键新基因的遗传鉴定

• 批准号: 8020998
• 负责人: LUANNE L PETERS
• 金额: $ 13.44万
• 依托单位: NEW YORK BLOOD CENTER
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-02-15 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8020998
• 关键词: Actins; Address; Affect; Alleles; Anemia; Area; Binding; Binding Proteins; Biological Assay; Blood; C-terminal; Cell membrane; Characteristics; Chromosome Mapping; Chromosomes, Human, Pair 8; Complex; Critiques; Defect; Densitometry; Erythrocytes; Erythropoiesis; Etiology; F-Actin; Genes; Genetic; Grant; Health; Hematopoietic stem cells; Heme; Hemolytic Anemia; High Pressure Liquid Chromatography; Human; Inherited; Investigation; Iron; Knowledge; Ligands; Literature; Location; Membrane; Messenger RNA; Methods; Mus; Mutation; Neonatal Anemia; Outcome; Pathogenesis; Pathology; Pathway interactions; Phenotype; Positioning Attribute; Principal Investigator; Proteins; Public Health; Published Comment; Research; Research Design; Research Methodology; Residual state; Sedimentation process; Spectrin; Suggestion; Testing; Thrombocytopenia; Vesicle; Work; Writing; alpha Spectrin; base; beta Spectrin; dimer; experience; gel electrophoresis; gene discovery; gene function; improved; iron metabolism; man; membrane skeleton; mouse model; mutant; new therapeutic target; novel; novel diagnostics; positional cloning; programs; protein 4.1; research study; response; uptake

DESCRIPTION (provided by applicant): Genetically defined mouse models have been instrumental in advancing our knowledge of the red blood cell (RBC) and the pathogenesis of inherited anemia. In this proposal, we will continue this powerful phenotype- driven approach to gene discovery and elucidation of mammalian gene function. The specific aims are: Aim 1. Positional cloning of scat and Nan. The recessive mouse mutation, scat (severe combined anemia and thrombocytopenia) and the dominant mutation, Nan (Neonatal anemia), display a severe anemia phenotype resulting from defects intrinsic to hematopoietic stem cells. Genetic mapping localized both scat and Nan to unique positions on chromosome 8 not associated with any previously identified mouse mutation. We hypothesize, therefore, that Nan and scat represent novel genes not previously recognized as significantly impacting blood formation. Here, we will (a) positionally clone the scat and Nan genes, (b) characterize the encoded mRNA and protein products, and (c) further characterize the scat and Nan hematological phenotypes. Aim 2. Characterization of novel mutant sph alleles. Five allelic recessive mouse mutations (spherocytosis, sph) with severe hemolytic anemia due to defects in the 1-spectrin gene (Spna1) have been previously described. We recently identified 2 additional sph alleles, sph3J and sph4J, that display unique phenotypic characteristics compared to the previously described alleles. Hence, we hypothesize that sph3J and sph4J disrupt novel interactions within the RBC membrane skeleton. We will (a) complete the phenotyping of sph3J and sph4J, (b) identify functional deficits incurred by the respective 1-spectrin mutations by characterizing interactions with known membrane skeleton components, and (c) identify novel spectrin interactions within the membrane. Relevance to Public Health: Inherited anemias in mouse and man share common etiologies. Thus, identifying the primary gene defect in mouse models of anemia has direct relevance to human health, will enhance our understanding of blood formation, and provide novel diagnostic and therapeutic targets for hematological pathologies.

描述（由申请人提供）：基因定义的小鼠模型有助于增进我们对红细胞（RBC）和遗传性贫血发病机制的了解。在本提案中，我们将继续这种强大的表型驱动方法来发现基因并阐明哺乳动物基因功能。具体目标是： 目标1. scat和Nan的定位克隆。隐性小鼠突变 scat（严重合并贫血和血小板减少症）和显性突变 Nan（新生儿贫血）显示出由造血干细胞固有缺陷引起的严重贫血表型。遗传图谱将 scat 和 Nan 定位在 8 号染色体上的独特位置，与任何先前发现的小鼠突变无关。因此，我们假设 Nan 和 scat 代表了以前未被认为会显着影响血液形成的新基因。在这里，我们将 (a) 按位置克隆 scat 和 Nan 基因，(b) 表征编码的 mRNA 和蛋白质产物，以及 (c) 进一步表征 scat 和 Nan 血液学表型。目标 2. 新型突变体 sph 等位基因的表征。先前已描述过由于 1-血影蛋白基因 (Spna1) 缺陷而导致严重溶血性贫血的五种等位基因隐性小鼠突变（球形红细胞增多症，sph）。我们最近鉴定了另外 2 个 sph 等位基因，sph3J 和 sph4J，与之前描述的等位基因相比，它们表现出独特的表型特征。因此，我们假设 sph3J 和 sph4J 破坏了红细胞膜骨架内的新相互作用。我们将 (a) 完成 sph3J 和 sph4J 的表型分析，(b) 通过表征与已知膜骨架成分的相互作用来识别由各自 1-血影蛋白突变引起的功能缺陷，以及 (c) 识别膜内新的血影蛋白相互作用。 与公共卫生的相关性：小鼠和人类的遗传性贫血具有共同的病因。因此，鉴定贫血小鼠模型中的主要基因缺陷与人类健康直接相关，将增强我们对血液形成的理解，并为血液病理学提供新的诊断和治疗靶点。