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.

Public Health Relevance

The proposal will use a novel technology to identify genes involved in the progression of prostate cancer. The proposed studies may rapidly identify new prostate cancer genes that can be targeted in future studies to inhibit cancer progression.

National Institute of Health (NIH)
National Cancer Institute (NCI)
Academic Research Enhancement Awards (AREA) (R15)
Project #
Application #
Study Section
Molecular Oncogenesis Study Section (MONC)
Program Officer
Okano, Paul
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Washington State University
Schools of Pharmacy
United States
Zip Code
Bii, Victor M; Trobridge, Grant D (2016) Identifying Cancer Driver Genes Using Replication-Incompetent Retroviral Vectors. Cancers (Basel) 8:
Rae, Dustin T; Collins, Casey P; Hocum, Jonah D et al. (2015) Modified Genomic Sequencing PCR Using the MiSeq Platform to Identify Retroviral Integration Sites. Hum Gene Ther Methods 26:221-7
Nalla, Arun K; Williams, Theodore F; Collins, Casey P et al. (2015) Lentiviral vector-mediated insertional mutagenesis screen identifies genes that influence androgen independent prostate cancer progression and predict clinical outcome. Mol Carcinog :
Hocum, Jonah D; Battrell, Logan R; Maynard, Ryan et al. (2015) VISA--Vector Integration Site Analysis server: a web-based server to rapidly identify retroviral integration sites from next-generation sequencing. BMC Bioinformatics 16:212
Rae, Dustin T; Hocum, Jonah D; Bii, Victor et al. (2015) A novel retroviral mutagenesis screen identifies prognostic genes in RUNX1 mediated myeloid leukemogenesis. Oncotarget 6:30664-74
Bii, Victor M; Rae, Dustin T; Trobridge, Grant D (2015) A novel gammaretroviral shuttle vector insertional mutagenesis screen identifies SHARPIN as a breast cancer metastasis gene and prognostic biomarker. Oncotarget 6:39507-20
Beard, Brian C; Adair, Jennifer E; Trobridge, Grant D et al. (2014) High-throughput genomic mapping of vector integration sites in gene therapy studies. Methods Mol Biol 1185:321-44
Schinke, Ellyn N; Bii, Victor; Nalla, Arun et al. (2014) A novel approach to identify driver genes involved in androgen-independent prostate cancer. Mol Cancer 13:120
Powers, John M; Trobridge, Grant D (2013) Identification of Hematopoietic Stem Cell Engraftment Genes in Gene Therapy Studies. J Stem Cell Res Ther 2013: