ETV4 mediates dosage-dependent prostate tumor initiation and cooperates with p53 loss to generate prostate cancer

The mechanisms underlying ETS-driven prostate cancer initiation and progression remain poorly understood due to a lack of model systems that recapitulate this phenotype. We generated a genetically engineered mouse with prostate-specific expression of the ETS factor, ETV4, at lower and higher protein dosages through mutation of its degron. Lower-level expression of ETV4 caused mild luminal cell expansion without histologic abnormalities and higher-level expression of stabilized ETV4 caused prostatic intraepithelial neoplasia (mPIN) with 100% penetrance within 1 week. Tumor progression was limited by p53-mediated senescence and Trp53 deletion cooperated with stabilized ETV4. The neoplastic cells expressed differentiation markers such as Nkx3.1 recapitulating luminal gene expression features of untreated human prostate cancer. Single-cell and bulk RNA-sequencing showed stabilized ETV4 induced a novel luminal-derived expression cluster with signatures of the cell cycle, senescence, and epit..., Mouse prostate digestion: Intraperitoneal injection of tamoxifen was administered in 8-week-old mice. 2 weeks after tamoxifen treatment, the mouse prostate was digested 1 hour with Collagenase/Hyaluronidase (STEMCELL, #07912), and then 30 minutes with TrypLETM Express Enzyme (Thermo Fischer, # 12605028) at 37°C to isolate single prostate cells. The prostate cells were stained with PE/Cy7 conjugated anti-mouse CD326 (EpCAM) antibody (BioLegend, 118216) and then, CD326 and EYFP double positive cells were sorted out by flow cytometry, which are luminal cells mainly from the anterior prostate and dorsal prostate. The mRNA or genomic DNA were extracted from these double-positive cells and then were used for ATAC-sequencing and RNA-sequencing analysis.
ATAC-seq and primary data processing: ATAC-seq was performed as previously described. Primary data processing and peak calling were performed using ENCODE ATAC-seq pipeline (https://github.com/kundajelab/atac_dnase_pipelines). Briefly, paired-e..., Please see the README document (\"README_Dataset-ETV4_mediates_dosage-dependent_prostate_tumor_initiation.txt\") and the accompanying published article: Dan Li, Yu Zhan, Naitao Wang, Fanying Tang, Cindy J. Lee, Gabriella Bayshtok, Amanda R. Moore, Elissa W.P. Wong, Mohini R. Pachai, Yuanyuan Xie, Jessica Sher, Jimmy L. Zhao, Makhzuna Khudoynazarova, Anuradha Gopalan, Joseph Chan, Ekta Khurana, Peter Shepherd, Nora M. Navone, Ping Chi, Yu Chen, ETV4 mediates dosage-dependent prostate tumor initiation and cooperates with p53 loss to generate prostate cancer. Science Advances 2023. Accepted. DOI: 10.5061/dryad.v41ns1s0s