FMS ONCOGENE--CSF-1 RECEPTOR

FMS癌基因--CSF-1受体

• 批准号: 3479640
• 负责人: CHARLES J SHERR
• 金额: $ 44.36万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-06-01 至 1995-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3479640
• 关键词: acute myelogenous leukemia; chemical structure function; colony stimulating factor; cytokine receptors; feline leukemia /sarcoma virus; fibroblasts; gene induction /repression; genetic manipulation; hematopoietic growth factor; human genetic material tag; human tissue; laboratory mouse; laboratory rabbit; laboratory rat; macrophage; molecular cloning; myelogenous leukemia; myeloid stem cell; nucleic acid probes; oncogenes; phosphorylation; protein kinase C; protooncogene; transfection; transposon /insertion element; viral leukemogenesis; virus genetics

The retroviral oncogene, v-fms, was acquired by genetic recombination between a feline leukemia virus (FeLV) and proto- oncogene sequences (c-fms) from normal cat cells. We demonstrated that the c-fms proto-oncogene encodes a receptor for the mononuclear phagocyte colony stimulating factor, CSF-1 (M-CSF). To date, this is the only system in which a hematopoietic growth factor and its receptor have both been cloned and characterized. Expression of c-fms at high levels in macrophages or after retroviral-mediated transfer into cultured fibroblasts does not lead to transformation, whereas v-fms transforms fibroblasts, CSF-1-dependent macrophages, and IL-3- dependent myeloid cell lines. An analysis of mutant c-fms and chimeric v-fms/c-fms genes suggested that two genetic alterations in c-fms are required to fully activate its oncogenic potential: (1) an activating mutation in the body of the gene that renders the receptor tyrosine kinase CSF-1-independent, and (2) elimination of a single C-terminal tyrosine residue (tyr969) that is likely to be a negative regulatory site of receptor phosphorylation. Additional chimeric and mutant receptor molecules will be used to pinpoint the site(s) of activating mutation(s), to identify sites of autophosphorylation, and to define putative target residues for protein kinase C phosphorylation that mediate receptor down modulation in response to phorbol esters. The structure and function of CSF-1 and its ability to transform cells by an autocrine mechanism when cotransfected with the c- fms gene will be studied. The ability of the CSF-1 receptor to program differentiative and proliferative responses will be evaluated after introducing the c-fms gene into committed myeloid precursors in vitro. Parallel approaches will assess the ability of the v-fms and CSF-1 genes to transform early myeloid progenitors in vitro and to contribute to leukemias in vivo after gene transfer into murine hematopoietic stem cells. The human c-fms and CSF-1 genes, both closely linked on human chromosome 5q, will be evaluated for specific rearrangements associated with acute myeloid leukemias. Studies at the genetic level will be complemented by biochemical approaches to identify defects in receptor function affecting CSF-1 induced kinase activity, receptor turnover, and down modulation. Our studies are likely to provide mechanistic information about normal hematopoiesis and to pinpoint defects in growth factor -- receptor interactions that contribute to leukemia.

逆转录病毒癌基因v-fms是通过遗传获得的， 猫白血病病毒（FeLV）和原 癌基因序列（c-fms）。 我们 表明c-fms原癌基因编码一种受体， 单核吞噬细胞集落刺激因子CSF-1 （M-CSF）。 迄今为止，这是唯一一个 造血生长因子及其受体都已被 克隆并鉴定。 c-fms在小鼠中的高水平表达 巨噬细胞或逆转录病毒介导的转移到培养的 成纤维细胞不会导致转化，而V-FMS 转化成纤维细胞、CSF-1依赖性巨噬细胞和IL-3- 依赖性骨髓细胞系。 突变型c-fms的分析和 嵌合v-fms/c-fms基因提示两种遗传 需要c-fms的改变来完全激活其致癌基因， 潜在的：（1）基因体内的激活突变， 使受体酪氨酸激酶CSF-1不依赖，和（2） 消除单个C-末端酪氨酸残基（tyr 969）， 可能是受体的负调控位点 磷酸化 另外的嵌合和突变受体 分子将被用来精确定位激活的位点， 突变，以鉴定自磷酸化位点，并确定 蛋白激酶C磷酸化的推定靶残基， 介导对佛波醇酯的受体下调。 CSF-1的结构、功能及其转化能力 细胞通过自分泌机制与c- 将研究fms基因。 CSF-1受体的能力， 程序分化和增殖反应将是 在将c-fms基因导入到已定型的 体外髓样前体细胞。 并行方法将评估 v-fms和CSF-1基因转化早期骨髓细胞的能力 在体外的祖细胞，并有助于白血病在体内后， 基因转移到小鼠造血干细胞中。 人类 c-fms和CSF-1基因，两者在人类染色体上紧密连锁 5 q，将评估与以下相关的特定重排： 急性骨髓性白血病 基因水平的研究将是 辅以生物化学方法， 受体功能影响CSF-1诱导的激酶活性， 受体转换和下调。 我们的研究很可能 提供有关正常造血的机制信息， 来确定生长因子-受体相互作用的缺陷， 会导致白血病