Rad9-Based Mouse Model of Prostate Carcinogenesis

基于 Rad9 的前列腺癌小鼠模型

• 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
• 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

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）： 当细胞暴露于损害DNA的物理或化学剂时，可能会随后发生有害作用，包括突变，癌症或死亡。但是，可以使用机制来修复损害，稳定基因组并中和有害影响。我们从酵母，小鼠和人类中分离并表征了一个称为rad9的进化保守基因，这对于维持基因组稳定性和促进细胞对DNA损伤的抗性至关重要。基于这些以及其他发现，我们研究了RAD9在致癌作用中的作用，并发现了与前列腺癌的重要联系。我们的结果包括相对于正常的人类前列腺PROC细胞，以及永生但非肿瘤的细胞系PWRE-1，RAD9在四种常用的前列腺癌细胞系中极度过表达CWR22，DU145，LNCAP和PC-3。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的肿瘤症的降低程度相关。此外，如果细胞经过设计以过量产生RAD9，则非果胶永生化的PWRE-1细胞会在裸鼠中形成异常的生长。此外，我们在339（45.1％）的人类前列腺腺癌中的153个中检测到了高RAD9蛋白水平，但在52个正常前列腺组织对照中的2个中，有2个蛋白质水平。相对较高的RAD9蛋白通常与更晚期疾病有关。因此，我们的结果表明RAD9在前列腺癌发生中起功能。该项目的重点是假设RAD9蛋白异常丰度导致前列腺癌。主要的实验目标是基于该器官中的RAD9的异常高表达来创建和表征前列腺癌变的新型小鼠模型。为此，我们拥有初步数据，表明已经构建了这样的鼠标线。该研究的具体目的是1）研究父母的前列腺组织，以及在前列腺中具有高水平RAD9的转基因小鼠，相对于自发性以及辐射诱导的前列腺组织异常和肿瘤发生的形成是年龄的函数； 2）确定高水平的RAD9与自发性以及电离辐射诱导的转移性前列腺癌之间的关系； 3）定义高水平RAD9介导前列腺疾病的能力与RAD9的射击功能的下游靶基因的表达与其他良好基因的表达，以与前列腺癌紧密相关的表达，开始了解RAD9介导的前列腺癌的机械基础。最后，该项目将提供一个独特的模型来定义前列腺致癌的分子机制，并作为阐明风险O环境致癌的体内系统。公共卫生相关性：该提案的重点是RAD9在介导前列腺致癌作用中的作用。目的是根据前列腺中的高水平的RAD9蛋白来设计和表征独特的小鼠模型，该模型表明对这种癌症的遗传易感性。将评估自发性和辐射诱导的前列腺疾病发展。因此，该项目很重要，因为结果可能会影响人类健康，无论是在治疗前列腺癌的治疗方面，以及了解遗传学和环境在癌变方面的相互作用。