Functional Genomic Dissection of Refractory Anemia

难治性贫血的功能基因组解析

• 批准号: 9113647
• 负责人: Benjamin Levine Ebert
• 金额: $ 44.38万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9113647
• 关键词: Acute leukemia; Alleles; Amino Acid Sequence; Amino Acids; Binding; Biochemical; Biochemistry; Biological Assay; Biology; Blood; Cell Count; Cell physiology; Cells; Chromosome Deletion; Chromosome abnormality; Chromosomes; Clonal Expansion; Complex; Congenital Disorders; Coupled; Cytogenetics; Development; Diamond-Blackfan anemia; Disease; Dissection; Drug resistance; Dysmyelopoietic Syndromes; Erythroid; Erythropoiesis; FDA approved; Funding; Genes; Genetic; Genetically Engineered Mouse; Genotype; Grant; Health; Hematopoietic stem cells; Human; Individual; Ineffective Hematopoiesis; Lesion; Link; Malignant Neoplasms; Methodology; Molecular; Molecular Abnormality; Mus; Mutagenesis; Mutate; Mutation; Oncogenic; Patients; Pharmaceutical Preparations; Phenotype; Post-Translational Protein Processing; Production; Protein-Serine-Threonine Kinases; Proteomics; Recurrence; Refractory anemias; Resistance development; Ribosomal Proteins; Role; Somatic Mutation; Syndrome; System; TP53 gene; Transplantation; Ubiquitination; base; beta catenin; casein kinase; chromosome 5q loss; clinical efficacy; clinical phenotype; combinatorial; functional genomics; gain of function; in vivo; interstitial; lenalidomide; member; mouse model; mutant; new therapeutic target; novel; phosphoproteomics; research study; response; ubiquitin ligase; ubiquitin-protein ligase

 DESCRIPTION (provided by applicant): Myelodysplastic syndrome (MDS) is characterized by ineffective hematopoiesis, most commonly of the erythroid lineage, resulting in a phenotype termed refractory anemia. Patients with MDS who have a heterozygous, interstitial deletion of chromosome 5q (Del (5q)), have a highly reproducible clinical phenotype, though the molecular basis of this phenotype was previously unknown. In the first funding period of this grant, we found that haploinsufficiency for RPS14, one of the genes within the common deleted region for del (5q) MDS, causes a block in erythropoiesis, the dominant clinical phenotype associated with this genetic abnormality, establishing a previously unrecognized link between del (5q) MDS and Diamond Blackfan anemia, a congenital disorder with a similar phenotype that is also caused by genetic inactivation of one allele of genes encoding ribosomal proteins. In the most recent funding period, we found that haploinsufficiency for CSNK1A1 causes elevated β-catenin levels, an increase in the number and function of hematopoietic stem cells, and sensitivity to the drug lenalidomide. Moreover, we identified recurrent, gain-of-function somatic mutations in CSNK1A1, the first gene within the Del (5q) common deleted region identified with recurrent somatic mutations. In the current proposal, we first aim to determine the mechanisms underlying clonal dominance of cells with CSNK1A1 heterozygous deletion or mutation using both murine models and phosphoproteomics. We next aim to determine how CSNK1A1 haploinsufficiency interacts with haploinsufficiency for other Del (5q) genes using a cellular barcoding system to perform competitive repopulation assays with multiple genotypes in a single mouse model. Finally, we will investigate the biochemistry of lenalidomide-dependent degradation by the CRL4CRBN ubiquitin ligase, using a novel saturation mutagenesis functional screen. These studies will inform the biology and treatment of Del (5q) MDS, the mechanisms underlying heterozygous deletions in cancer more broadly, and the development of novel therapeutics targeting ubiquitin ligase activity.

 描述（由适用提供）：骨髓增生综合征（MDS）的特征是无效的造血症，最常见的是红斑谱系，导致一种称为难治性贫血的表型。患有杂合子5q（DEL（5Q））杂合，间质缺失的MDS患者具有高度可重现的临床表型，尽管以前未知该表型的分子基础。在这笔赠款的第一个资金期间，我们发现RPS14的单倍不使人不适，这是DEL（5Q）MD的普通已删除区域中的基因之一，导致促红细胞生成的障碍，这是与此遗传异常相关的主导临床表型，与先前未识别的del（5q）Mdnemment Angity Is Nection And Angunity Is Nection Andemant Andement and Mondem Mand and Mand Mand and Anginia Mondem and Mondem and Monginal Angine Anement a Mondial and Anginape由一个编码核糖体蛋白的基因等位基因的遗传失活引起。在最近的资金期间，我们发现CSNK1A1的单倍不足会导致β-catenin水平升高，造血干细胞的数量和功能增加以及对药物为lenalidomide的敏感性。此外，我们确定了CSNK1A1的复发性，功能获得的体细胞突变，这是DEL（5Q）常见缺失区域内的第一个基因，该基因鉴定出具有复发性体细胞突变的鉴定。在当前的建议中，我们首先旨在确定使用鼠模型和磷蛋白质组学使用CSNK1A1杂合缺失或突变的细胞基础机制。接下来，我们的目的是确定CSNK1A1单倍度不足与其他DEL（5Q）基因使用细胞条形码系统在单个小鼠模型中使用多个基因型进行竞争性重新分析测定法。最后，我们将使用一种新型的饱和诱变功能筛选来研究CRL4CRBN泛素连接酶的Lenalidomide依赖性降解的生物化学。这些研究将为DEL（5Q）MDS的生物学和治疗提供信息，这是癌症中杂合缺失的基本机制，以及针对泛素连接酶活性的新型治疗的发展。