TYROSINE KINASE FUSION IN HEMATOLOGIC MALIGNANCY

该项目将表征酪氨酸激酶融合在 小鼠恶性血液病的发病机制 白血病和淋巴瘤的模型。我们实验室最近克隆了 几种酪氨酸激酶的融合将为这些 学习。我们已经克隆了TEL-PDGFRβ融合及其相关变异体 与t(5；12)慢性粒单核细胞白血病(CMML)相关，以及一种新的 TEL-ABL融合与急性髓系白血病相关。我们的合作者， 克隆了一个与高级别相关的NPM-ALK酪氨酸激酶融合基因 淋巴瘤。我们将建立白血病和淋巴瘤的小鼠模型 TEL-PDGFRbeta、TEL-ABL和NPM-ALK融合。 与慢性粒细胞白血病(CML)的bcr-abl一样，TEL-PDGFRβ，TEL-ABL ABL和NPM-ALK可能通过结构性激活授予恶性表型 PDGFRbeta和ALK的酪氨酸激酶结构域。在具体目标1中，我们 将表征TEL和PDGFRβ在患有(I)的患者中的参与 CMML与t(5；12)易位，(Ii)正常核型CMML，(Iii) 骨髓增生异常综合征的其他FAB亚型，(Iv)髓系化生和 骨髓纤维化，(V)，急性单核细胞白血病，和(Vi)血液病 伴有12p13细胞遗传学异常的恶性肿瘤。这些研究将会有所帮助 TELL和PDGFRβ在血液病发病机制中的作用 恶性，并可能提供对功能域的洞察 在转变活动中很重要。在具体目标2中，转型 TELL-PDGFRbeta和TELL-ABL融合基因的活性将在 稳定表达的哺乳动物细胞系。TEL-PDGFRβ和TELL-PDGFRβ的表达 TELL-ABL融合将通过免疫印迹和体外激酶证实 化验。具体目标1和2将为具体目标3提供基础 将评估TEL-PDGFRbeta、TEL-ABL和NPM的转型潜力- ALK融合在白血病小鼠骨髓移植模型中的应用 淋巴瘤。对移植小鼠的评估将包括详细的 组织病理学检查；白细胞分类和 免疫表型分析确定血液病家系受累 恶性；白血病细胞和白血病细胞系的分析 移植小鼠前病毒整合和表达的证据 融合蛋白。这些研究将确定TEL的作用- PDGFRβ、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，并可能为更有效的治疗提供洞察力 酪氨酸激酶融合介导的血液病恶性。此外， 这些研究将为免疫治疗方法提供基础。 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.