Mutagenesis Screen for Prostate Cancer

前列腺癌诱变筛查

基本信息

批准号：
8574446
负责人：
GRANT D TROBRIDGE
金额：
$ 41.78万
依托单位：
WASHINGTON STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-16 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8574446
关键词：
Androgens Bacteria Biological Markers Cancer Etiology Cancer Patient Candidate Disease Gene Castration Cell Culture Techniques Cells Cessation of life Charcoal Chromosomes Databases Detection Disease Progression Distant Environment Epithelial-Stromal Communication Future Gene Expression Gene Expression Profile Genes Growth Human In Vitro Injection of therapeutic agent LNCaP Libraries Location Malignant neoplasm of prostate Mediating Metastatic Prostate Cancer Molecular Mus Mutagenesis Mutate Mutation Neoplasm Metastasis Oncogenes PC3 cell line Parents Plasmids Process Prostate Proteome Proteomics Protocols documentation Proviruses Reverse Transcriptase Polymerase Chain Reaction Risk Screening for Prostate Cancer Serum Shuttle Vectors Site Technology Tumor Suppressor Genes Tumor Suppressor Proteins United States Validation Vascularization Viral Xenograft Model Xenograft procedure anticancer research base cancer genome cancer type candidate identification castration resistant prostate cancer clinically relevant comparative genomic hybridization deep sequencing design gene discovery in vitro Model in vivo knock-down male men new technology new therapeutic target novel outcome forecast overexpression promoter prostate cancer cell prostate cancer model public health relevance small hairpin RNA tumor tumor growth tumor progression vector

项目摘要

DESCRIPTION (provided by applicant): Prostate cancer is the second most common cause of cancer related deaths in men, yet relatively little is known about the molecular mechanisms of disease progression. Identifying genes that are involved in this process is important to identify new therapeutic targets. Here we propose to establish a novel retroviral shuttle vector mutagenesis technology to identify genes that drive prostate cancer progression. Comparative genome hybridization (CGH), transcriptome deep sequencing, and proteomics have emerged as technologies for prostate cancer gene discovery. However, distinguishing the causal mutations from the entire cancer genome/transcriptome/proteome remains a challenge. We have developed a novel mutagenesis vector that allows efficient high-throughput identification of candidate dysregulated genes. In this approach, integrated lentiviral shuttle vectors mutagenize target cells by dysregulating the expression of nearby genes. Vector provirus:chromosome junctions are rescued as plasmids in bacteria, and these plasmids are sequenced to identify the chromosomal location of each vector provirus. This identifies nearby candidate dysregulated genes that promote cancer growth and/or progression. Our shuttle vector technology overcomes a severe technical limitation of previous retroviral mutagenesis screens, inefficient PCR-based detection. We will take advantage of an established LNCaP xenograft prostate cancer model to establish our novel technology. Using this technology we expect to rapidly identify novel genes that drive prostate cancer in vivo. These genes can be targeted in future studies to inhibit prostate cancer or used as biomarkers. In future studies our approach may be used for other types of cancer to accelerate the pace of cancer research with broad potential impact.

描述（由申请人提供）：前列腺癌是男性癌症相关死亡的第二大最常见原因，但对疾病进展的分子机制知之甚少。识别参与此过程的基因对于识别新的治疗靶标很重要。在这里，我们建议建立一种新型的逆转录病毒媒介诱变技术，以鉴定驱动前列腺癌进展的基因。比较基因组杂交（CGH），转录组深测序和蛋白质组学已成为前列腺癌基因发现的技术。但是，将因果突变与整个癌症基因组/转录组/蛋白质组区分开是一个挑战。我们已经开发了一种新型的诱变载体，该诱变载体可以有效地对候选基因失调的高通量鉴定。在这种方法中，综合的慢病毒穿梭载体通过失调附近基因的表达来诱变靶细胞。载体原病毒：染色体连接在细菌中被挽救为质粒，这些质粒被测序以鉴定每个载体病毒的染色体位置。这确定了附近的候选基因失调，促进癌症的生长和/或进展。我们的航天飞机矢量技术克服了以前的逆转录病毒诱变筛查的严重技术限制，基于PCR的检测效率低下。我们将利用已建立的LNCAP异种移植前列腺癌模型来建立我们的新技术。使用这项技术，我们期望快速识别在体内驱动前列腺癌的新型基因。这些基因可以在以后的研究中旨在抑制前列腺癌或用作生物标志物。在未来的研究中，我们的方法可能用于其他类型的癌症，以加速具有广泛潜在影响的癌症研究的速度。