Rad9-Based Mouse Model of Prostate Carcinogenesis

基于 Rad9 的前列腺癌小鼠模型

基本信息

批准号：
7728289
负责人：
HOWARD B. LIEBERMAN
金额：
$ 24.05万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-01 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7728289
关键词：
Address Aftercare Age Androgen Receptor Animals Apoptotic Benign Prostatic Hypertrophy Binding Biopsy Cancerous Carcinogen exposure Carcinogens Cell Cycle Checkpoint Cell Cycle Progression Cell Line Cells Cessation of life Characteristics Chemical Agents Chemicals Complement Consensus Sequence DNA Damage DU145 Data Defect Development Disease Engineering Environment Environmental Carcinogens Fission Yeast Frequencies Future Gamma Rays Gene Targeting Genes Genetic Genetic Predisposition to Disease Genetic Recombination Genetic Transcription Genome Genome Stability Genomics Goals Grant Growth Health Human Human Cloning Hyperplasia Injection of therapeutic agent Investigation Ionizing radiation LNCaP Lead Left Link Liver Location Lung Maintenance Malignant Neoplasms Malignant neoplasm of prostate Mediating Mediator of activation protein Metastatic Prostate Cancer Microtomy Modeling Molecular Morphology Mus Mutation Neoplasm Metastasis Neoplasms Nude Mice Organ Other Genetics PC3 cell line Play Predisposition Primary Neoplasm Prostate Prostate Adenocarcinoma Prostatic Diseases Prostatic Intraepithelial Neoplasias Prostatic Neoplasms Proteins RNA Radiation Radiation Interaction Relative (related person)Research Resistance Risk Role Severities Severity of illness Small Interfering RNA Staging System TP53 gene Testing Time Tissues Transactivation Transgenic Mice Transgenic Organisms Tumorigenicity Up-Regulation Vascular Endothelial Growth Factor Receptor-1 Yeasts advanced disease base bone cancer cell cancer site cancer therapy carcinogenesis cell growth cellular engineering environmental carcinogenesis genetic element immortalized cell in vivo irradiation lymph nodes male mouse model novel outcome forecast overexpression promoter prostate carcinogenesis public health relevance rad9 protein repaired research study tool tumorigenesis tumorigenic

项目摘要

DESCRIPTION (provided by applicant): When cells are exposed to physical or chemical agents that damage DNA, deleterious effects can ensue, including mutation, cancer or death. However, mechanisms are available to repair the damage, stabilize the genome and neutralize harmful effects. We isolated and characterized an evolutionarily conserved gene called Rad9, from yeast, mouse and human, which is critical for maintaining genomic stability and promoting cellular resistance to DNA damage. Based on these as well as other findings, we examined the role of Rad9 in carcinogenesis, and found a significant link in particular to prostate cancer. Our results include the observation that Rad9 is dramatically overexpressed in four commonly used prostate cancer cell lines, CWR22, DU145, LNCaP and PC-3, relative to normal human prostate PrEC cells and the immortalized but nontumorigenic cell line PWRE-1. We also found that the degree to which siRNA reduces Rad9 protein levels correlates with the extent of decrease in tumorigenicity of these cells after injection into nude mice. Moreover, nontumorigenic immortalized PWRE-1 cells can form aberrant growths in nude mice if the cells are engineered to overproduce Rad9. In addition, we detected high Rad9 protein levels in 153 of 339 (45.1%) human prostate adenocarcinomas, but just low abundance in 2 of 52 normal prostate tissue controls. Relatively higher levels of Rad9 protein were in general associated with more advanced disease. Our results thus indicate that Rad9 plays a functional role in prostate carcinogenesis. This project focuses on the hypothesis that aberrantly high abundance of Rad9 protein leads to prostate cancer. The major experimental goal is to create and characterize a novel mouse model of prostate carcinogenesis, based on aberrant high expression of Rad9 specifically in that organ. To this end, we have preliminary data indicating that such a line of mouse has been constructed. Specific aims of the study focus on 1) examining prostate tissue of parental, as well as transgenic mice with high levels of Rad9 in prostate with respect to spontaneous as well as radiation-induced formation of prostate tissue abnormalities and tumorigenesis as a function of age; 2) determine the relationship between high levels of Rad9 and spontaneous as well as ionizing radiation-induced metastatic prostate cancer; and 3) defining the relationship between the ability of high levels of Rad9 to mediate prostate disease and expression of downstream target genes of the transactivity function of Rad9 as well as expression of other genes well-established to be intimately involved in prostate cancer, to begin to understand the mechanistic basis for Rad9 mediated prostate carcinogenesis. In the end, this project will provide a unique model to define molecular mechanisms of prostate carcinogenesis, and serve as an in vivo system for elucidating risk o environmental carcinogenesis. PUBLIC HEALTH RELEVANCE: This proposal focuses on the role of Rad9 in mediating prostate carcinogenesis. The goal is to engineer and characterize a unique mouse model demonstrating genetic predisposition to this kind of cancer, based on high levels of Rad9 protein in prostate. The spontaneous as well as radiation-induced development of prostate disease will be assessed. As such, this project is important since the results could impact on human health, both in terms of treatment for prostate cancer and understanding the interplay of genetics and the environment with respect to carcinogenesis.

描述（由申请人提供）：当细胞暴露于损伤 DNA 的物理或化学试剂时，就会产生有害影响，包括突变、癌症或死亡。然而，有一些机制可以修复损伤、稳定基因组并消除有害影响。我们从酵母、小鼠和人类中分离并鉴定了一种名为 Rad9 的进化保守基因，该基因对于维持基因组稳定性和促进细胞对 DNA 损伤的抵抗力至关重要。基于这些以及其他发现，我们检查了 Rad9 在致癌作用中的作用，并发现了其与前列腺癌之间的显着联系。我们的结果包括观察到，相对于正常人前列腺 PrEC 细胞和永生化但非致瘤细胞系 PWRE-1，Rad9 在四种常用前列腺癌细胞系 CWR22、DU145、LNCaP 和 PC-3 中显着过度表达。我们还发现，siRNA 降低 Rad9 蛋白水平的程度与注射到裸鼠后这些细胞致瘤性降低的程度相关。此外，如果非致瘤性永生化 PWRE-1 细胞被设计为过量产生 Rad9，那么这些细胞可能会在裸鼠体内形成异常生长。此外，我们在 339 例人类前列腺腺癌中的 153 例 (45.1%) 中检测到高 Rad9 蛋白水平，但在 52 例正常前列腺组织对照中的 2 例中检测到低丰度。相对较高水平的 Rad9 蛋白通常与更晚期的疾病相关。因此，我们的结果表明 Rad9 在前列腺癌发生中发挥功能性作用。该项目的重点是假设异常高丰度的 Rad9 蛋白会导致前列腺癌。主要实验目标是基于 Rad9 在该器官中的异常高表达，创建并表征前列腺癌发生的新型小鼠模型。为此，我们有初步数据表明这样的小鼠线已经被构建出来。该研究的具体目标集中在1)检查亲本前列腺组织以及前列腺中Rad9水平高的转基因小鼠的前列腺组织自发形成以及辐射诱导的前列腺组织异常形成和随年龄变化的肿瘤发生； 2) 确定高水平的Rad9与自发性以及电离辐射诱发的转移性前列腺癌之间的关系； 3) 确定高水平的 Rad9 介导前列腺疾病的能力与 Rad9 反式活性功能的下游靶基因的表达以及已确定与前列腺癌密切相关的其他基因的表达之间的关系，以开始了解 Rad9 介导的前列腺癌发生的机制基础。最终，该项目将提供一个独特的模型来定义前列腺癌发生的分子机制，并作为阐明环境癌发生风险的体内系统。公共健康相关性：该提案重点关注 Rad9 在介导前列腺癌发生中的作用。我们的目标是基于前列腺中高水平的 Rad9 蛋白，设计并表征一种独特的小鼠模型，证明这种癌症的遗传易感性。将评估前列腺疾病的自发发展和辐射诱发的发展。因此，该项目非常重要，因为其结果可能对人类健康产生影响，无论是在前列腺癌的治疗方面还是在了解遗传学和环境与致癌作用之间的相互作用方面。