Studies on the molecular mechanisms of chromosome instability and development of MDS/AML

染色体不稳定与MDS/AML发生的分子机制研究

基本信息

批准号：
11670982
负责人：
YAMASHITA Takayuki
金额：
$ 2.11万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

Fanconi anemia (FA) is an autosomal recessive disease characterized by bone marrow failure, which is predisposed to development of myelodysplastic syndrome and acute myeloid leukemia, chromosomal instability and hypersensitivity to DNA damaging agents such as mitomycin C.There are at least eight genetically distinct groups (A, B, C, D1, D2, E, F and G) in FA, and thus far six genes (FANCA, C, D2, E, F and G) have been cloned. Multiple FA proteins encoded by these genes have been shown to cooperate in a molecular pathway, which is critical in the maintenance of genomic stability in hematopoietic stem cells. Our purpose is to clarify molecular pathogenesis in Japanese FA patients, based on the new understanding of pathophysiology of FA.In order to study structure-function relationship of FANCA, we established multiple transformants stably expressing various mutant FANCA proteins in FANCA(-) cells and are analyzing several functions of these cells. Also, we detected some FANCA-interacting proteins using yeast two-hybrid and co-immunopecipitation and are studying functional significance of these interactions. We developed a diagnostic method to identify abnormalities of FA genes, using protein analyses and functional complementation of patient cells with retroviral transduction of wildtype FA genes. We detected some mutations characteristic to the Japanese population. Our data indicate that analyses of FANCD2 ubiquitination is a reliable diagnostic tool for FA.

Fanconi anemia (FA) is an autosomal recessive disease characterized by bone marrow failure, which is predisposed to development of myelodysplastic syndrome and acute myeloid leukemia, chromosomal instability and hypersensitivity to DNA damaging agents such as mitomycin C.There are at least eight genetically distinct groups （a，b，c，d1，d2，e，f和g）在FA中，到目前为止六个基因（Fanca，C，D2，E，F和G）已被克隆。这些基因编码的多种FA蛋白已显示在分子途径中合作，这对于维持造血干细胞的基因组稳定性至关重要。我们的目的是根据对FA的病理生理学的新理解来阐明日本FA患者的分子发病机理，以研究FANCA的结构 - 功能关系，我们建立了多种转化器，稳定地表达了Fanca（ - ）细胞中各种突变的Fanca蛋白正在分析这些细胞的几个功能。同样，我们使用酵母两杂交和共免疫沉淀检测到了一些FANCA相互作用的蛋白质，并正在研究这些相互作用的功能意义。我们开发了一种诊断方法，使用蛋白质分析和逆转录病毒FA基因倒流转导的患者细胞的功能互补，以鉴定FA基因的异常。我们检测到了日本人口特征的一些突变。我们的数据表明，对FANCD2泛素化的分析是FA的可靠诊断工具。