ISOLATION OF A SECOND WILMS TUMOR SUPPRESSOR GENE

第二个 Wilms 肿瘤抑制基因的分离

• 批准号: 6140623
• 负责人: Bernard E. Weissman
• 金额: $ 3.88万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-06-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6140623
• 关键词: Wilms' tumor; athymic mouse; chimeric proteins; chromosome translocation; complementary DNA; gene mutation; human genetic material tag; human tissue; neoplasm /cancer genetics; nucleic acid sequence; oligonucleotides; polymerase chain reaction; single strand conformation polymorphism; synthetic peptide; tissue /cell culture; transfection; tumor suppressor genes; tumor suppressor proteins

DESCRIPTION: (adapted from the investigator's abstract) The identification of human tumor suppressor genes has led to new insights into the mechanisms of human cancer development. Some of the first tumor suppressor genes were identified through studies of pediatric malignancies including the RB and WTI genes. In the case of Wilms' tumors, further investigations have led to the discovery of other potential tumor suppressor genes on chromosomes 11, 16 as well as an unmapped familial form. In a complementary fashion, he has taken a functional approach by using a biological assay, tumor suppression, to map the locations of functional tumor suppressor genes via monochromosome transfer. He has now narrowed the location of a second Wilms' tumor suppressor gene, WT2, to an approximately 350 kb region on 11p15.5. Other studies have placed translocations in Beckwith-Weidemann Syndrome patients and loss of heterozygosity for Wilms tumor samples in this same region. In addition, this region of the human genome contains genes subject to inactivation by genomic imprinting. Intriguingly, many Wilms' tumors show a loss of imprinting for genes in 11p15.5 leading to either their inactivation or increased expression. The role of these epigenetic events such as imprinting in Wilms' tumor and other human cancer development remains unknown. Therefore, the isolation and characterization of the WT2 tumor suppressor gene would constitute a major advance in understanding these influences. During the last funding period, he developed a PAC/BAC/PI contig across the WT2 tumor suppressor gene region and began the identification of candidate genes in the area. In this competitive renewal application, he proposes to isolate the WT2 gene and characterize its status in the development of Wilms' tumor. In Specific Aim A, he will identify as many genes as possible for the WT2 tumor suppressor region by solution hybrid capture and analysis of genomic DNA sequence. In Specific Aim B, he will screen each candidate gene for correlative expression with tumor suppression in a microcell hybrid model system. He also will search for genomic alterations in primary tumor samples and examine the expression pattern in normal tissues. He hopes to limit the number of strong candidate genes by these criteria to five or less. In the last specific aim, he will identify the WT2 gene by screening for mutations and loss of expression in primary tumor samples. He will also transfer the gene into the G401 cell line to demonstrate functional tumor suppressor activity. The availability of the WT2 gene will broaden the understanding of tumor suppressor gene functions, provide important clues about the process of normal mammalian tissue development including genomic imprinting and impact upon treatment and detection of human cancer.

描述：（改编自研究者的摘要）鉴定 人类肿瘤抑制基因的研究带来了对其机制的新见解 人类癌症的发展。 一些最早的肿瘤抑制基因是 通过对儿科恶性肿瘤（包括 RB 和 WTI 基因。 就维尔姆斯氏肿瘤而言，进一步的研究导致 在 11 号染色体上发现其他潜在的肿瘤抑制基因， 16 以及未映射的家族形式。 以一种互补的方式，他 通过使用生物测定、肿瘤抑制等功能性方法， 通过单色体绘制功能性肿瘤抑制基因的位置图 转移。 他现在已经缩小了第二个维尔姆斯肿瘤的位置 抑制基因 WT2 位于 11p15.5 上约 350 kb 的区域。 其他 研究已将易位置于贝克威斯-魏德曼综合征患者身上 同一区域中肾母细胞瘤样本的杂合性丢失。 在 此外，人类基因组的这个区域包含受 通过基因组印记灭活。 有趣的是，许多肾母细胞瘤表现出 11p15.5 基因印记丢失导致其失活 或表达增加。 这些表观遗传事件的作用，例如 维尔姆斯氏瘤和其他人类癌症发展中的印记仍然存在 未知。 因此，WT2肿瘤的分离和表征 抑制基因将构成理解这些的重大进步 影响。 在上一次资助期间，他开发了 PAC/BAC/PI 重叠群跨越 WT2 肿瘤抑制基因区域并开始 识别该区域的候选基因。 在这场竞争激烈的更新中 在申请中，他建议分离 WT2 基因并表征其状态 维尔姆斯氏瘤的发展。 在具体目标 A 中，他将确定为 通过解决方案获得 WT2 肿瘤抑制区域的尽可能多的基因 基因组 DNA 序列的混合捕获和分析。 在具体目标 B 中，他 将筛选每个候选基因与肿瘤的相关表达 微单元混合模型系统中的抑制。 他也会寻找 原发性肿瘤样本中的基因组改变并检查表达 正常组织中的模式。 他希望限制强势候选人的数量 根据这些标准，基因数为 5 个或更少。 在最后一个具体目标中，他将 通过筛选突变和表达缺失来鉴定 WT2 基因 原发性肿瘤样本。 他还将把基因转移到G401细胞中 线以证明功能性肿瘤抑制活性。 可用性 WT2基因的研究将拓宽对抑癌基因的理解 功能，为正常哺乳动物的生命过程提供重要线索 组织发育，包括基因组印记和对治疗的影响 和人类癌症的检测。