TUMOR SUPPRESSOR LOCI IN MESOTHELIOMA

间皮瘤中的肿瘤抑制基因座

• 批准号: 2683616
• 负责人: JONATHAN Alfred FLETCHER
• 金额: $ 28.43万
• 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/2683616
• 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：（改编自研究者的摘要）恶性 间皮瘤是一种与石棉相关的肿瘤，越来越多的公众 健康问题，并带来巨大的诊断和治疗 挑战。至少 65% 的间皮瘤有中层缺失 6 号染色体长臂的一部分，值得注意的是，这与 该区域在乳腺癌、白血病、非霍奇金病中被非随机删除 淋巴瘤、骨肉瘤和前列腺癌。弗莱彻博士和同事 假设 6q16.3-q21 区域的抑癌基因是 大多数间皮瘤中缺失和/或突变，并且它们还 假设该基因在非间皮瘤中具有广泛的相关性 肿瘤发生。弗莱彻博士将通过精细绘图来解决这些假设 通过定位、分离和表征 6q 缺失区域 平衡的细胞遗传学重排会中断该区域，并且通过 评估间皮瘤和非肿瘤中的候选抑癌基因 具有 6q 缺失的间皮瘤。映射将分两次进行 阶段。最初，弗莱彻博士将绘制染色体的广泛区域图 使用已建立的方法在细胞系中的 30 个原发性间皮瘤中发现 6 个长臂 大型 YAC 的 10 名成员 FISH 小组，间隔为 2-10 兆碱基。这 映射阶段将定义关键删除区域，同时 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 使用细菌人工染色体 (BAC) FISH 检测间皮瘤 以 500 kb 间隔跨越关键删除区域的面板。这些 研究可能会从细胞遗传学角度揭示表观杂合子和 纯合缺失将进一步缩小关键缺失范围 地区。 删除映射将与 mega-YAC FISH 协调 平衡细胞遗传学断点的定位和克隆 共识删除区域内的重排。 筛选 候选抑癌基因将通过直接cDNA完成 使用包含关键的 100-250 kb BAC DNA 序列进行选择 缺失区域和细胞遗传学断点。候选抑癌基因 基因将根据拷贝数（FISH）进行评估， 失活点突变（SSCP/测序）和表达（Northern 印迹）在原发性间皮瘤、间皮瘤细胞系和非 具有已知 6q 缺失的间皮瘤癌症。长期目标 包括评估 6q 肿瘤抑制基因作为潜在的 间皮瘤的诊断标志物及其生物学评价 通过功能研究对该基因的元素进行研究。