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

• 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
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670982/
• 关键词: Fanconi anemia; chromosome instability; myelodysplastic syndrome; leukemia; ファンユニ貧血; 急性骨髄性白血病; リン酸化; 蛋白リン酸化酵素

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贫血(FA)是一种以骨髓衰竭为特征的常染色体隐性遗传病，易发生骨髓增生异常综合征和急性髓系白血病，染色体不稳定，对丝裂霉素C等DNA损伤剂超敏。FA至少有8个基因群(A、B、C、D_1、D_2、E、F和G)，迄今已克隆出6个基因(FANCA、C、D_2、E、F和G)。由这些基因编码的多个FA蛋白已被证明在一条分子途径上相互协作，这在维持造血干细胞基因组稳定性方面是至关重要的。我们的目的是在对FA的病理生理学有了新的认识的基础上，阐明日本FA患者的分子发病机制。为了研究FANCA的结构与功能的关系，我们在FANCA(-)细胞中建立了稳定表达各种突变的FANCA蛋白的多个转化子，并对这些细胞的几种功能进行了分析。此外，我们还利用酵母双杂交和免疫共沉淀检测到了一些FANCA相互作用的蛋白，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，通过逆转录病毒转导野生型FA基因，利用蛋白质分析和患者细胞的功能互补来识别FA基因的异常。我们检测到了一些日本人特有的突变。我们的数据表明，FANCD2泛素化分析是一种可靠的FA诊断工具。

项目成果

Oda,T.,Yamashita,T. et al.: "A novel SH2 domain-containing protein, HSH2, may link tyrosine kinase signaling to cpc42-regulated kinase Ackl in hematopoiete cells."Blood. 96. 78a (2000)

小田，T.，山下，T.

Kupfer, G., Yamashita, T., et al.: "A patient-derived mutant of Fanconi anemia protein, FANCA, is defective in nuclear accumulation"Exp Hematol. 27. 587-593 (1999)

Kupfer, G.、Yamashita, T. 等人：“源自患者的 Fanconi 贫血蛋白突变体 FANCA 在核积累方面存在缺陷”Exp Hematol。

Yamashita, T., Nakahata, T.: "Current Know ledge on the Pathogenesis of Fanconi Anemia : from genes to phenotypes"Int J. Hematol. (印刷中). (2001)

Yamashita, T., Nakahata, T.：“范可尼贫血发病机制的当前知识：从基因到表型”Int J. Hematol（印刷中）。

Yamashita T and Nakahata T.: "Current knowledge on the pathophysiology of Fanconi anemia : from genes to phenotypes."Int J Hematol. (in press).

Yamashita T 和 Nakahata T.：“范可尼贫血病理生理学的最新知识：从基因到表型。”Int J Hematol。

Futaki,M.,Yamashita,T. et al.: "The IVS4+4A to T mutation of the Fanani anemia gene FANCC is not associated with a sever phenotype in Japanese patients."Blood. 95. 1493-1498 (2000)

二木，M.，山下，T.