IDENTIFICATION OF A 12P PROSTATE TUMOR SUPPRESSOR GENE

12P 前列腺肿瘤抑制基因的鉴定

• 批准号: 6633919
• 负责人: ADAM S KIBEL
• 金额: $ 12.15万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-05-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6633919
• 关键词: DNA methylation; complementary DNA; gene deletion mutation; genetic mapping; genetic promoter element; human genetic material tag; human tissue; laboratory mouse; loss of heterozygosity; metastasis; neoplasm /cancer genetics; neoplastic growth; plasmids; prostate neoplasms; tumor suppressor genes

DESCRIPTION (Applicant's Description): The two objectives of this application are to support my training as a physician scientist and to identify novel tumor suppressor gene(s) with functional significance in prostate carcinoma. This award will facilitate my transition to independent investigator by providing detailed molecular laboratory training under the mentorship of Dr. Paul Goodfellow, the Director of the Cancer Genetics Research Program at the School of Medicine. Dr. Goodfellow is ideally suited to be my mentor because his research into the genetic determinants of endometrial tumorigenesis closely parallels the research proposed in this application; therefore, all techniques described in this application have been used in his laboratory. Importantly, he has previously successfully mentored several physician scientists. A didactic program in molecular biology will complement the intellectual environment provided by Dr. Goodfellow's lab, the Cancer Genetics Research Program, and the Division of Urology. I have recently reported the discovery of a 1-2Mb homozygous deletion (HZD) at 1 2 p12-13 in metastatic prostate cancer specimens. Frequent loss of heterozygosity (LOH) of the same region is frequently seen in tumors. The smallest common region of LOH encompasses the location of the HZD and narrows the consensus deletion region to approximately 600kb. The frequency and regional specificity of these deletions (LOH and HZD) point to a prostate cancer suppressor gene at this site. The experimental rationale and methods proposed to clone this 12pl2-13 prostate tumor suppressor gene are as follows: (I) Define the minimal candidate region at 12pl2-13. An overlapping HZD in a second prostate cancer specimen would narrow the region of interest. I will use computational and laboratory screening techniques to identify candidate genes in this region and then determine if candidates are HZD in a panel of prostatic tumors. (II) Investigate candidate genes inactivating genetic events. If these genes contribute to prostate tumorigenesis, inactivating genetic events will be present in the remaining prostate carcinoma specimens. Cancer specimens will be interrogated for mutations and promotor methylation. (III) Functional demonstration of tumor suppression. If loss of function of the 12p12-13 gene contributes to tumor formation, reintroduction of the wild- t y p e , but not mutant cDNA into prostate cell lines will suppress tumorigenesis. The identification of a novel tumor suppressor would provide a target for better prostate cancer therapy and/or more accurate prognostic information for men newly diagnosed with the disease.

描述（申请人的描述）：本申请的两个目标 是为了支持我作为一名医师科学家的培训并识别新颖的 在前列腺癌中具有功能意义的肿瘤抑制基因。 该奖项将有助于我过渡到独立调查员 在博士的指导下提供详细的分子实验室培训 Paul Goodfellow，癌症遗传学研究项目主任 医学院。 Goodfellow 博士非常适合成为我的导师，因为 他对子宫内膜肿瘤发生的遗传决定因素的研究 与本申请中提出的研究非常相似；因此，所有 本申请中描述的技术已在他的实验室中使用。 重要的是，他之前曾成功指导过几位医生 科学家。 分子生物学教学计划将补充 Goodfellow 博士的癌症遗传学实验室提供的智力环境 研究计划和泌尿科。 我最近在以下网址报告了 1-2Mb 纯合缺失 (HZD) 的发现： 1 2 p12-13 在转移性前列腺癌标本中。 经常丢失 同一区域的杂合性（LOH）在肿瘤中很常见。 这 LOH 的最小公共区域包含 HZD 的位置并缩小 共识删除区域约为 600kb。 频率和 这些缺失（LOH 和 HZD）的区域特异性指向前列腺 该位点的抑癌基因。 实验原理和方法 提议克隆此12pl2-13前列腺抑癌基因如下： (I) 定义 12pl2-13 处的最小候选区域。 重叠的 HZD 第二个前列腺癌标本将缩小感兴趣区域。 我会 使用计算和实验室筛选技术来识别候选人 该区域中的基因，然后确定候选者是否是一组中的 HZD 前列腺肿瘤。 （二）研究失活遗传的候选基因 事件。 如果这些基因有助于前列腺肿瘤的发生，那么使其失活 遗传事件将存在于剩余的前列腺癌样本中。 将检查癌症样本的突变和启动子甲基化。 （三）抑制肿瘤的功能论证。 如果丧失功能 12p12-13 基因有助于肿瘤形成、野生动物的重新引入 型，但突变型cDNA进入前列腺细胞系会抑制 肿瘤发生。 新型肿瘤抑制因子的鉴定将为肿瘤抑制提供靶点 更好的前列腺癌治疗和/或更准确的预后信息 新诊断出患有这种疾病的男性。