Translational genomics

转化基因组学

• 批准号: 7068947
• 负责人: Thomas E. Ried
• 金额: --
• 依托单位: DIVISION OF CLINICAL SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7068947
• 关键词: aneuploidy; biomedical automation; breast neoplasm /cancer diagnosis; breast neoplasms; carcinogenesis; cervical /vaginal smear; cervix neoplasms; chromosome aberrations; chromosome translocation; comparative genomic hybridization; computer program /software; cytogenetics; diagnosis design /evaluation; diagnostic tests; fluorescent in situ hybridization; gene expression profiling; genetic markers; imaging /visualization /scanning; mass screening; neoplasm /cancer diagnosis; neoplasm /cancer genetics; proteomics

CGH analyses of human tumors have revealed that chromosomal aberrations result in genomic imbalances specific for different tumor from diverse tissue types. Furthermore, these changes define discrete steps in the progression of epithelial tumors. More than 90% of cervical carcinomas carry extra copies of chromosome 3 and virtually all diploid breast cancers show a gain of chromosome 1q. Also, gains of chromosome 3q precede copy number increases of chromosomes 5p and 1, and loss of chromosome 2 during the genesis of cervical carcinomas. It is therefore conducive to apply the visualization of these recurring and specific chromosomal aberrations to complement and enhance the cytomorphological diagnosis of human cancers and their precursor lesions. This can be achieved using interphase cytogenetics with fluorescently tagged DNA probes that recognize specific chromosomal target regions directly in interphase cells. We have focused on three applications: Identification of the progressive potential of cervical intraepithelial neoplasia based on the detection of extra copies of chromosome 3 and 5 in thinprep PAP-smears. Diagnosis and prognostication of suspicious breast lesions following the detection of chromosomal aneuploidies in fine needle aspirates. Visualization of specific chromosomal aneuploidies in cytokeratin positive epithelial cells isolated from the peripheral blood of breast cancer patients.These projects are accompanied by the improvement of procedures for the preparation of cytological specimens for interphase cytogenetics, by the development of directly labeled, multicolor probe cocktails for relevant chromosomal regions, oncogenes, and tumor suppressor genes (supported by a CRADA with Vysis, Inc.), and by the development of semi-automated or automated microscope hardware and imaging software for fluorescent spot counting. We could show that the visualization of chromosomal aneupoloidies in cytological preparations (fine needle aspirates from the breast and monolayer PAP-smears) is highly specific and sensitive tests for the diagnosis of cancer and premalignant precursor lesions. For instance, the detection of genomic copy number alterations of the human telomerase gene hTERT (which maps to chromosome 3q26) serves as an independent genetic marker for the diagnosis of cervical dysplasia. Other translational research efforts include the use of parallel gene expression profiling for predicting therapy response in rectal carcinomas, and the use of proteomics to identify early markers for the recurrence of colon tumors after surgery.

对人类肿瘤的CGH分析表明，染色体异常导致了来自不同组织类型的不同肿瘤的基因组不平衡。此外，这些变化定义了上皮性肿瘤进展的离散步骤。超过90%的宫颈癌携带额外的3号染色体拷贝，几乎所有二倍体乳腺癌都显示出1Q染色体的增加。在宫颈癌的发生过程中，染色体3q的获得先于5p和1染色体的拷贝数增加和2号染色体的丢失。因此，应用这些复发的和特定的染色体异常的可视化有助于补充和加强对人类癌症及其前驱病变的细胞形态诊断。这可以使用间期细胞遗传学和荧光标记的DNA探针来实现，这些探针直接识别间期细胞中的特定染色体靶区。我们将重点放在三个应用上：基于检测ThinPrep PAP涂片中额外的3号和5号染色体拷贝来识别宫颈上皮内瘤变的进展潜力。在细针抽吸物中检测到染色体非整倍体后可疑乳腺病变的诊断和预测。从乳腺癌患者外周血中分离的细胞角蛋白阳性的上皮细胞中特定的染色体非整倍体的可视化。这些项目伴随着制备间期细胞遗传学的细胞学标本的程序的改进，通过开发相关染色体区域、癌基因和肿瘤抑制基因的直接标记的多色探针鸡尾酒(由与Vosis，Inc.的CRADA支持)，以及开发用于荧光斑点计数的半自动或自动显微镜硬件和成像软件。我们可以证明，在细胞学准备(乳腺细针抽吸物和单层PAP涂片)中染色体非整倍体的可视化是诊断癌症和癌前病变的高度特异和敏感的测试。例如，检测人类端粒酶基因hTERT(位于染色体3q26)的基因组拷贝数变化可作为诊断宫颈异型增生的独立遗传标记。其他翻译研究工作包括使用平行基因表达谱来预测直肠癌的治疗反应，以及使用蛋白质组学来确定结肠肿瘤手术后复发的早期标志物。