IDENTIFICATION OF A 12P PROSTATE TUMOR SUPPRESSOR GENE

描述（申请人的描述）：本申请的两个目的 是为了支持我作为一名医生科学家的训练， 在前列腺癌中具有功能意义的肿瘤抑制基因。 这个奖项将有助于我过渡到独立调查员， 提供详细的分子实验室培训的指导下博士。 Paul Goodfellow，癌症遗传学研究项目主任， 医学院。 古德费洛博士是我的理想导师 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密切相关;因此，所有 本申请中描述的技术已经在他的实验室中使用。 重要的是， 他曾成功指导过几位医生， 科学家 分子生物学的教学计划将补充 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。 我最近报告了在1999年发现的1- 2 Mb纯合缺失（HZD）。 1 2转移性前列腺癌标本中的p12-13。 经常失去 同一区域的杂合性（洛）在肿瘤中常见。 的 洛缺失的最小共同区域包括HZD的位置， 将共有缺失区扩增至约600 kb。 的频率和 这些缺失（洛和HZD）的区域特异性指向前列腺 癌抑制基因在这个位置。 实验原理和方法 提出克隆该12 p12 -13前列腺肿瘤抑制基因的方法如下： (I)在12 pl 2 -13处定义最小候选区域。 一个重叠的HZD 第二前列腺癌样本将缩小感兴趣区域。 我会 使用计算和实验室筛选技术来确定候选人 基因，然后确定候选人是否是HZD在一组 前列腺肿瘤 (II)研究使遗传失活的候选基因 事件 如果这些基因有助于前列腺肿瘤的发生， 遗传事件将存在于剩余的前列腺癌样本中。 将询问癌症标本的突变和启动子甲基化。 (III)肿瘤抑制的功能演示。 如果功能丧失， 12 p12 -13基因有助于肿瘤形成，野生型的重新引入， t y p e，但不能将突变的cDNA导入前列腺细胞系中， 肿瘤发生 一种新的肿瘤抑制因子的鉴定将提供一个靶点， 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人。

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.