Background: Alterations (alt) in TP53, PTEN and RB1 TSGs have been identified in some prostate cancers. Preclinical data suggest that cooperative loss of 2 or more TSGs drives development of more aggressive disease. Methods: We identified men who underwent targeted exome sequencing (DFCI PROFILE) of CSPC (localized [L] and M1 presentation) and CRPC tissue samples. Biomarker(BM)-positive(+) was defined as copy number loss or deleterious mutation of ≥1 TSG (TP53, PTEN or RB1). For pts presenting with L-CSPC, Kaplan-Meier method estimated time from biopsy to PSA relapse/metastasis/death (EFS), CRPC, and death (OS). Cox model assessed association of BM status and outcomes, adjusted for age, stage and Gleason score in multivariate analyses (MVA). Time from ADT start for M1-CSPC to CRPC and death was also estimated. For M1-CRPC, duration on 1^st line CRPC therapy and time from CRPC to death was estimated, adjusting for age, volume and location of metastasis in MVA. Association of cumulative BM+ hits (0 vs 1 vs 2 vs 3) and outcomes was assessed. Results: For BM+ frequencies see table. L-CSPC with BM+ had a shorter EFS (median 2.6 years, HR 1.95, 95% CI 1.22-3.13) and time to CRPC (HR 3.36, 95% CI 1.01-11.16). MVA confirmed association with EFS (HR 1.84, p = 0.029). More gene hits lead to greater risk of relapse (MVA; 1 vs 0 hit: HR 1.75, p = 0.05; 2/3 vs 0: HR 2.74, p = 0.04). None of the 43 M1-CSPC pts who were BM-neg had died with median follow-up of 3.3 yrs; BM+ 4-year OS was 64%. Only 4 (8%) of the CRPC cohort (n = 48) were BM-neg and with a median follow-up 4.1 years, only 1 had died (5.2 yrs). Cumulative TSG loss was associated with shorter duration on 1^st line therapy in MVA (3 vs 0/1 hit: HR 2.86, p = 0.013). Conclusions: Deleterious TP53, PTEN and RB1 variants are associated with increased risk of relapse (M0) and death (M1) in CSPC and shorter duration on 1^st line therapy in CRPC. BM-neg in CRPC is rare but may represent a subset of pts with very good prognosis. Poorer outcomes are seen with cumulative gene hits across cohorts.