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：(改编自调查人员的摘要)恶性 间皮瘤是一种与石棉相关的肿瘤，其发病率越来越高。 健康问题，并造成巨大的诊断和治疗 挑战。至少65%的间皮瘤有中段缺失 6号染色体长臂的一部分，值得注意的是 区域在乳腺癌、白血病、非何杰金氏病中的非随机缺失 淋巴瘤、骨肉瘤和前列腺癌。弗莱彻博士和同事 假设6q16.3-q21区域的肿瘤抑制基因是 在大多数间皮瘤中缺失和/或突变，它们还 假设该基因在非间皮瘤中具有广泛相关性 肿瘤发生学。弗莱彻博士将通过精细的映射来解决这些假设 通过作图、分离和鉴定6q缺失区 平衡的细胞遗传学重排，中断这一区域，并通过 间皮瘤和非间皮瘤中候选抑癌基因的评价 6q缺失的间皮瘤。映射将分两部分执行 阶段。最初，弗莱彻博士将绘制大范围的染色体区域图 30个细胞系中的6个长臂间皮瘤使用已建立的 由10个成员组成的巨型YAC鱼群，间隔2-10兆基数。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，弗莱彻博士将对同一组进行重新评估 应用细菌人工染色体(BAC)FISH技术治疗间皮瘤 以500 kb的间隔跨越关键缺失区域的面板。这些 研究可能在细胞遗传学上揭示明显的杂合子和 纯合缺失将进一步缩小关键缺失的范围 区域。删除图谱将与巨型YAC鱼进行协调 细胞遗传学平衡断裂点的定位和克隆 协商一致删除区域内的重新安排。筛选 候选的肿瘤抑制基因将由直接的cdna完成。 使用100-250 kb的BAC DNA序列进行选择 缺失区和细胞遗传学断裂点。候选肿瘤抑制因子 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 Blotting)在原发间皮瘤、间皮瘤细胞系和非 已知6q缺失的间皮瘤癌症。长期目标 包括评价6q抑癌基因作为潜在的 间皮瘤的诊断标记物及其生物学评价 通过功能研究确定了该基因的组成成分。