TUMOR SUPPRESSOR LOCI IN MESOTHELIOMA

间皮瘤中的肿瘤抑制基因座

• 批准号: 2390897
• 负责人: JONATHAN Alfred FLETCHER
• 金额: $ 27.44万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-04-05 至 1999-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2390897
• 关键词: artificial chromosomes; bacteria; chromosome walking; clinical research; complementary DNA; cytogenetics; fluorescent in situ hybridization; gene deletion mutation; genetic mapping; human subject; human tissue; mesothelioma; molecular oncology; neoplasm /cancer genetics; northern blottings; nucleic acid sequence; tumor suppressor genes

DESCRIPTION: (adapted from the investigator's abstract) Malignant mesothelioma is an asbestos-associated neoplasm which is a growing public health concern and which poses tremendous diagnostic and therapeutic challenges. At least 65% of mesotheliomas have deletion of the mid- portion of the chromosome 6 long arm, and it is notable that this same region is deleted nonrandomly in breast cancer, leukemia, non-Hodgkin's lymphoma, osteosarcoma, and prostate cancer. Dr. Fletcher and coworkers hypothesize that a tumor suppressor gene in the 6q16.3-q21 region is deleted and/or mutated in the majority of mesotheliomas, and they also hypothesize that this gene has broad relevance in non-mesothelioma tumorigenesis. Dr. Fletcher will address these hypotheses by fine mapping of the 6q deletion region, by mapping, isolating, and characterizing balanced cytogenetic rearrangements that interrupt this region, and by evaluating candidate tumor suppressor genes in mesotheliomas and in non- mesothelioma tumors with 6q deletions. Mapping will be performed in two phases. Initially, Dr. Fletcher will map a broad region of the chromosome 6 long arm in 30 primary mesotheliomas in cell lines using an established 10-member FISH panel of mega-YACs spaced at 2-10 megabase intervals. This phase of mapping will define the critical deletion region while simultaneously evaluating the possibility of additional deletion regions. In the second phase Dr. Fletcher will re-evaluate the same group of mesotheliomas using a bacterial artificial chromosome (BAC) FISH panel spanning the critical deletion region at 500 kb intervals. These studies may reveal cytogenetically in apparent heterozygous and homozygous deletions which will further narrow the critical deletion region. The deletion mapping will be coordinated with mega-YAC FISH mapping and cloning of breakpoints for balanced cytogenetic rearrangements within the consensus deletion region. Screening for candidate tumor suppressor genes will then be accomplished by direct cDNA selection using 100-250 kb BAC DNA sequences containing the critical deletion region and a cytogenetic breakpoint. Candidate tumor suppressor genes will be evaluated with a respect to copy number (FISH), inactivating point mutations (SSCP/sequencing), and expression (Northern blotting) in primary mesotheliomas, mesothelioma cell lines, and in non- mesothelioma cancers with known 6q deletions. Long-term objectives include evaluation of the 6q tumor suppressor gene as a potential diagnostic marker in mesothelioma and evaluation of the biological elements of this gene through functional studies.

描述：（改编自调查员的摘要）恶性 间皮瘤是一种与石棉相关的肿瘤，它是一个日益增长的公众 健康问题，并带来极大的诊断和治疗性 挑战。至少有65％的间皮瘤删除了中间 染色体6长臂的一部分，值得注意的是 在乳腺癌，白血病，非霍奇金的乳腺癌中删除了非随机删除 淋巴瘤，骨肉瘤和前列腺癌。弗莱彻博士和同事 假设在6q16.3-q21区域中的肿瘤抑制基因是 在大多数间皮瘤中删除和/或突变，它们也 假设该基因在非间皮瘤中具有广泛的相关性 肿瘤发生。 Fletcher博士将通过精细的映射来解决这些假设 通过映射，隔离和表征6Q删除区域 平衡的细胞遗传重排，中断该区域，并通过 评估间皮瘤和非候选肿瘤抑制基因 间皮瘤肿瘤具有6Q缺失。映射将在两次 阶段。最初，弗莱彻博士将绘制染色体的广泛区域 使用已建立 由10人的巨型鱼板以2-10兆瓦的间隔间隔。这 映射阶段将定义关键缺失区域，而 同时评估额外删除的可能性 地区。在第二阶段，弗莱彻博士将重新评估同一组 使用细菌人造染色体（BAC）鱼的间皮瘤 面板以500 kb的间隔跨越关键缺失区域。这些 研究可能在明显的杂合和 纯合删除将进一步缩小关键缺失 地区。 删除映射将与大型鱼协调 平衡细胞遗传学的断点的映射和克隆 共识缺失区域内的重排。 筛选 然后，候选肿瘤抑制基因将通过直接cDNA完成 使用包含关键的100-250 kb BAC DNA序列的选择 缺失区域和细胞遗传学断点。候选肿瘤抑制剂 基因将以尊重拷贝数（鱼）的尊重进行评估， 灭活点突变（SSCP/测序）和表达（北 在原发性间皮瘤，间皮瘤细胞系和非 - 间皮瘤癌症患有已知6Q缺失。长期目标 包括评估6Q肿瘤抑制基因作为潜力 间皮瘤中的诊断标记和生物学评估 通过功能研究，该基因的要素。