ISOLATION OF A SECOND WILMS TUMOR SUPPRESSOR GENE

描述：(改编自调查人员的摘要)身份 对人类肿瘤抑制基因的研究使人们对其机制有了新的认识 人类癌症的发展。一些最早的肿瘤抑制基因是 通过对包括Rb和Rb在内的儿童恶性肿瘤的研究确定 WTI基因。在Wilms肿瘤的案例中，进一步的调查导致了 11号染色体上其他潜在肿瘤抑制基因的发现， 16以及一个未映射的家族性形式。作为补充，他已经 采取了一种功能性的方法，使用生物测试，肿瘤抑制， 利用单染色体定位功能性抑癌基因 调职。他现在已经缩小了第二个Wilms瘤的位置 抑制基因WT2位于11p15.5，大小约为350 kb。其他 研究发现Beckwith-Weidemann综合征患者存在易位 和在同一区域的Wilms肿瘤样本的杂合性丢失。在……里面 此外，人类基因组的这一区域包含受 通过基因组印迹来灭活。耐人寻味的是，许多Wilms‘s肿瘤显示出 11p15.5中的基因印记丢失导致其失活 或者增加了表达。这些表观遗传事件的作用，如 肾母细胞瘤和其他人类癌症发展过程中的印记 未知。因此，WT2肿瘤的分离和鉴定 抑制基因将在理解这些方面取得重大进展。 影响。在上一个资助期内，他开发了PAC/BAC/PI 重叠群穿过WT2抑癌基因区域并开始 该地区候选基因的鉴定。在这场竞争性的更新中 应用，他建议分离WT2基因并鉴定其状态 在肾母细胞瘤的发展过程中。在具体目标A中，他将确定为 通过解决WT2肿瘤抑制区可能的多个基因 基因组DNA序列的杂交捕获和分析。在具体目标B中，他 将筛选每个候选基因与肿瘤相关的表达 微细胞混合模型系统中的抑制。他还将寻找 原发肿瘤标本的基因组改变及其表达检测 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，基因只有五个或更少。在最后一个具体目标中，他将 通过筛查WT2基因突变和表达缺失来鉴定WT2基因 原发肿瘤样本。他还将把基因转移到G401细胞中 展示功能性肿瘤抑制活性的线路。可得性 WT2基因的发现将拓宽人们对抑癌基因的认识 功能，为正常哺乳动物的进化过程提供了重要线索 组织发育包括基因组印记及其对治疗的影响 以及人类癌症的检测。

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.