TYROSINE KINASE FUSION IN HEMATOLOGIC MALIGNANCY

该项目将描述酪氨酸激酶融合在 血液恶性肿瘤的发病机制，通过小鼠的发展， 白血病和淋巴瘤的模型。 我们的实验室最近克隆了 几个酪氨酸激酶融合，这将提供这些基础， 问题研究 我们克隆了TEL-PDGFR β融合蛋白及其相关变体 与t（5;12）慢性粒单核细胞白血病（CMML）相关， TEL-ABL融合与急性髓系白血病相关 我们的合作者， 已克隆了与高恶性肿瘤相关的JNPM-ALK酪氨酸激酶融合蛋白， 淋巴瘤 我们将开发白血病和淋巴瘤的小鼠模型， TEL-PDGFR β、TEL-ABL和NPM-ALK融合体。 与慢性髓细胞性白血病（CML）中的BCR-ABL一样，TEL-PDGFR β、TEL-PDGFR β和TEL-ABL也是如此。 ABL和NPM-ALK可能通过组成性激活赋予恶性表型 PDGFR β和ALK的酪氨酸激酶结构域。 具体目标1， 将描述TEL和PDGFR β在以下患者中的参与：（i） CMML和t（5;12）易位，（ii）具有正常核型的CMML，（iii） 骨髓增生异常综合征的其他FAB亚型，（iv）髓样化生和 骨髓纤维化，（v）急性单核细胞白血病，和（vi）血液学 恶性肿瘤伴12 p13细胞遗传学异常。 这些研究将有助于 阐明TEL和PDGFR β在血液病发病机制中的作用， 恶性肿瘤，并可能提供洞察功能域， 在转化活动中很重要。 在具体目标2中， TEL-PDGFR β和TEL-ABL融合基因的活性将在 稳定转染的哺乳动物细胞系。 TEL-PDGFR β的表达和 TEL-ABL融合将通过免疫印迹和体外激酶 测定。 具体目标1和2将为具体目标3提供基础 将评估TEL-PDGFR β、TEL-ABL和NPM的转化潜力- 白血病小鼠骨髓移植模型中的ALK融合， 淋巴瘤 移植小鼠的评估将包括详细的 组织病理学检查;白色血细胞分类和 免疫表型分析以确定血液系统疾病谱系参与 恶性肿瘤;和分析白血病细胞和白血病细胞系， 移植小鼠的前病毒整合和表达的证据， 融合蛋白。 这些研究将描述电话的作用- PDGFR β、TEL-ABL和NPM-ALK融合蛋白在白血病发病机制中的作用 和淋巴瘤，并可能提供更有效的治疗， 酪氨酸激酶融合介导血液恶性肿瘤。 此外，委员会认为， 这些研究将为免疫学方法提供基础， 白血病和淋巴瘤。

This project will characterize the role of tyrosine kinase fusions in pathogenesis of hematologic malignancy, through development of murine models of leukemia and lymphoma. Our laboratory has recently cloned several tyrosine kinase fusions which will provide the basis for these studies. We have cloned the TEL-PDGFRbeta fusion and related variants associated with t(5;12) chronic myelomonocytic leukemia (CMML), and a novel TEL-ABL fusion associated with acute myeloid leukemia. Our collaborator, has cloned athe NPM-ALK tyrosine kinase fusion associated with high grade lymphomas. We will develop murine models of leukemia and lymphoma using the TEL-PDGFRbeta, TEL-ABL and NPM-ALK fusions. As with BCR-ABL in chronic myelogenous leukemia (CML), TEL-PDGFRbeta, TEL- ABL and NPM-ALK may confer a malignant phenotype by constitutive activation of the tyrosine kinase domains of PDGFRbeta and ALK. In Specific Aim 1 we will characterize involvement of TEL and PDGFRbeta in patients with (i) CMML and t(5;12) translocation, (ii) CMML with normal karyotype, (iii) other FAB subtypes of myelodysplastic syndrome, (iv) myeloid metaplasia and myelofibrosis, (v), acute monocytic leukemias, and (vi) hematologic malignancy with 12p13 cytogenetic abnormalities. These studies will help to clarify the role of TEL and PDGFRbeta in pathogenesis of hematologic malignancy, and may provide insight into functional domains which are important in transforming activity. In Specific Aim 2, transforming activity of the TEL-PDGFRbeta and TEL-ABL fusion gene will be confirmed in stably transfected mammalian cell lines. Expression of TEL-PDGFRbeta and TEL-ABL fusions will be confirmed by immunoblotting and in vitro kinase assays. Specific Aims 1 and 2 will provide the basis for Specific Aim 3 will assess transforming potential of the TEL-PDGFRbeta, TEL-ABL and NPM- ALK fusions in murine bone marrow transplant models of leukemia and lymphoma. Evaluation of transplanted mice will include detailed histopathologic examination; white blood cell differential and immunophenotype analysis to determine lineage involvement of hematological malignancy; and analysis of leukemic cells and leukemic cell lines from transplanted mice for evidence of proviral integration and expression of the fusion protein. These studies will characterize the role of the TEL- PDGFRbeta, TEL-ABL and NPM-ALK fusion proteins in pathogenesis of leukemia and lymphoma, and may provide insight into more effective therapy of hematologic malignancy mediated by tyrosine kinase fusions. Furthermore, these studies will provide a foundation for immunotherapeutic approaches to leukemia and lymphoma.