Functional Genomic Dissection of Refractory Anemia

难治性贫血的功能基因组解析

• 批准号: 7022794
• 负责人: TODD R. GOLUB
• 金额: $ 32.22万
• 依托单位: DANA-FARBER CANCER INST
• 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 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7022794
• 关键词: CD34 molecule; RNA interference; biotechnology; bone marrow; cell differentiation; cell proliferation; chromosomes; clinical research; comparative genomic hybridization; drug discovery /isolation; drug screening /evaluation; dyserythropoietic anemia; erythroid stem cell; functional /structural genomics; gene deletion mutation; gene expression; gene expression profiling; high throughput technology; human subject; human tissue; microarray technology; molecular cloning; phenotype; small molecule

DESCRIPTION (provided by applicant): Myelodysplastic syndrome (MDS) has been long recognized as a collection of diseases characterized by ineffective hematopoiesis associated with impairment of differentiation and intra-medullary apoptosis. Moreover, patients with MDS often progress to acute myeloid leukemia (AML). The pathogenesis of MDS remains obscure, and effective therapies are lacking. One challenge is that MDS represents a diverse collection of diseases, categorized by often vague clinical criteria. One exception to this, however, is the 5q syndrome, which has distinct clinical characteristics and is associated with deletions of the long arm of chromosome 5. The disease gene for this syndrome has yet to be identified. We therefore propose here an ambitious genomics-based approach to the cloning of the 5q- syndrome disease gene, and to the discovery of small molecules that circumvent the differentiation block that is characteristic of MDS. In Aim 1, we will perform a systematic, RNA interference-based functional genomic screen in CD34+ primary hematopoietic progenitor cells to identify those genes in the critically deleted region of 5q that recapitulate the differentiation defect characteristic of MDS. In addition, we will perform fine-mapping of the 5q region using high density custom microarray comparative genomic hybridization (CGH) in order to identify patients with previously unrecognized, small deletions. Furthermore, we will utilize .a novel single-molecule sequencing method to resequence the candidate genes on 5q for mutations. In Aim 2, we will perform a high throughput gene expression-based small molecule screen using the GE-HTS method developed in our laboratory. Using this approach, we expect to identify small molecules (or drugs) capable of promoting the expansion and differentiation of hematopoietic progenitors. Such compounds would be valuable both as tool compounds with which to dissect the biology of MDS, and as potential starting points for the development of new therapies of MDS.

描述（由申请人提供）： 骨髓增生异常综合征（MDS）是一组以造血功能低下、分化障碍和髓内细胞凋亡为特征的疾病。 此外，MDS患者通常进展为急性髓性白血病（AML）。MDS的发病机制尚不清楚，缺乏有效的治疗方法。一个挑战是MDS代表了多种疾病的集合，通常通过模糊的临床标准进行分类。然而，5 q综合征是一个例外，它具有独特的临床特征，与5号染色体长臂缺失有关。这种综合征的致病基因尚未确定。因此，我们在这里提出了一个雄心勃勃的基于基因组学的方法来克隆5 q综合征疾病基因，并发现小分子，绕过分化块，这是MDS的特征。在目标1中，我们将在CD 34+原代造血祖细胞中进行系统的、基于RNA干扰的功能基因组筛选，以鉴定5 q关键缺失区域中概括MDS分化缺陷特征的那些基因。此外，我们将使用高密度定制微阵列比较基因组杂交（CGH）对5 q区域进行精细定位，以识别先前未识别的小缺失患者。此外，我们将利用一种新的单分子测序方法对5 q上的突变候选基因进行重新测序。在目标2中，我们将使用我们实验室开发的GE-HTS方法进行基于基因表达的高通量小分子筛选。使用这种方法，我们期望鉴定能够促进造血祖细胞扩增和分化的小分子（或药物）。这样的化合物作为剖析MDS生物学的工具化合物和作为开发MDS新疗法的潜在起点都是有价值的。