ISOLATION OF A SECOND WILMS TUMOR SUPPRESSOR GENE

第二个 Wilms 肿瘤抑制基因的分离

• 批准号: 6266679
• 负责人: Bernard E. Weissman
• 金额: $ 6.65万
• 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/6266679
• 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：（改编自研究者摘要） 对人类肿瘤抑制基因的研究使人们对肿瘤发生的机制有了新的认识。 人类癌症的发展。 一些最早的肿瘤抑制基因是 通过儿科恶性肿瘤研究确定，包括RB和 WTI基因。 在肾母细胞瘤的情况下，进一步的调查导致 在11号染色体上发现了其他潜在的肿瘤抑制基因， 16、一个没有血缘关系的家族。 作为一种补充，他 通过使用生物测定，肿瘤抑制， 通过单染色体定位功能性肿瘤抑制基因 转移 他现在已经缩小了第二个肾母细胞瘤的位置 抑制基因WT 2定位于11p15.5上约350 kb的区域。 其他 研究表明贝-魏二氏综合征患者 在同一区域的肾母细胞瘤样本中存在杂合性缺失。 在 此外，人类基因组的这一区域包含受以下影响的基因： 通过基因组印记失活。 有趣的是，许多Wilms肿瘤显示出 11p15.5中基因印记的丢失导致其失活 或增加表达。 这些表观遗传事件的作用， 在肾母细胞瘤和其他人类癌症发展中的印记仍然存在 未知 因此，WT 2肿瘤的分离和表征 抑制基因将构成理解这些的重大进展， 影响。 在上一个资助期间，他开发了一个PAC/BAC/PI 在WT 2肿瘤抑制基因区域的重叠群，并开始 确定该地区的候选基因。 在这次竞争激烈的续约中 应用，他建议分离WT 2基因并表征其状态 在威尔姆斯肿瘤的发展中。 在具体目标A中，他将确定为 尽可能多的基因为WT 2肿瘤抑制区的解决方案 杂交捕获和基因组DNA序列分析。 在特定目标B中，他 筛选出与肿瘤相关的候选基因 微单元混合模型系统中的抑制。 他还将寻找 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。 他希望限制强有力的候选人的数量 根据这些标准，他将基因减少到5个或更少。在最后一个具体目标中，他将 通过筛选WT 2基因的突变和表达缺失， 原发性肿瘤样品。 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。 的可用性 对WT 2基因的研究将拓宽对抑癌基因的认识 功能，提供有关正常哺乳动物的过程的重要线索， 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。