Discovery of novel T cell oncogenes by using a functional retroviral cDNA library screen.

通过使用功能性逆转录病毒 cDNA 文库筛选发现新型 T 细胞癌基因。

• 批准号: nhmrc : 384144
• 负责人: Dr David Izon
• 金额: $ 46.17万
• 依托单位: St. Vincent's Institute of Medical Research
• 依托单位国家: 澳大利亚
• 项目类别: NHMRC Project Grants
• 财政年份: 2006
• 资助国家: 澳大利亚
• 起止时间: 2006-01-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://researchdata.edu.au/discovery-novel-t-library-screen/98148
• 关键词: Discovery; novel; cell; oncogenes; using

T cells mature in an organ called the thymus which is located on top of the heart. Blood borne T cell precursors enter the thymus after being resident in the bone marrow. T cell leukaemia is a disease where a blood cell that is committed to becoming a T cell is blocked from maturing into a functional cell. Instead, the leukaemic immature T cell uncontrollably divides to make endless non-functional copies of itself. As a result, normal functional T cells are outcompteted and the immune system is crippled. Patients generally die due to opportunistic infection. The molecular causes of T cell leukaemia are slowly being discovered. Up to 50% of all human T cell leukaemias overexpress SCL-TAL-1. Other T cell leukaemia-causing genes (oncogenes) include Ras and Notch. Current leukaemia treatments include chemotherapy and bone marrow transplants but even these fail ~30% of the time. Consequently, all T cell oncogenes need to be discovered so that disease-specific treatments can be generated. This proposal will utlise a functional retroviral cDNA library screen to uncover novel T cell lineage commitment genes and T cell oncogenes. This will be accomplished by constructing a coloured [GFP] cDNA library (a library of genes) that will be transfected (inserted) into immature T cells that cannot develop down the T cell pathway owing to the lack of a crucial gene (Rag-1). The T cell oncogene Ras and the T cell lineage commitment gene Notch can move cells past the Rag-1 block. If there is a gene in the cDNA library that can compensate for the lack of Rag-1 and allow the cells to mature we will detect it using high speed flow cytometryic cell sorting (like sieving weevils from flour very quickly). Once we find this cell we will isolate the gene using the colour tag. The potential oncogenes uncovered will provide the foundation for next generation drug development that targets each leukaemia based on its cause.

T细胞在一个叫做胸腺的器官中成熟，胸腺位于心脏顶部。血液携带的T细胞前体在驻留在骨髓中之后进入胸腺。T细胞白血病是一种疾病，其中致力于成为T细胞的血细胞被阻止成熟为功能细胞。相反，白血病未成熟的T细胞不受控制地分裂，产生无休止的无功能的自身拷贝。结果，正常功能的T细胞被淘汰，免疫系统受损。患者通常死于机会性感染。T细胞白血病的分子原因正在慢慢被发现。高达50%的人T细胞白血病过表达SCL-TAL-1。其他导致T细胞白血病的基因（癌基因）包括Ras和Notch。目前的白血病治疗包括化疗和骨髓移植，但即使这些治疗也有30%的失败率。因此，需要发现所有T细胞癌基因，以便产生疾病特异性治疗。本研究将利用功能性逆转录病毒cDNA文库筛选新的T细胞谱系定型基因和T细胞癌基因。这将通过构建彩色[GFP] cDNA文库（基因文库）来实现，该文库将被转染（插入）到由于缺乏关键基因（Rag-1）而不能沿T细胞途径发育的未成熟T细胞中。T细胞癌基因Ras和T细胞谱系定型基因Notch可以使细胞通过Rag-1阻断。如果cDNA文库中有一个基因可以弥补Rag-1的缺失并使细胞成熟，我们将使用高速流式细胞术细胞分选（如快速筛选面粉中的象鼻虫）检测它。一旦我们找到这个细胞，我们将使用颜色标签分离基因。发现的潜在致癌基因将为下一代药物开发提供基础，这些药物将根据其病因靶向每种白血病。