Background: Anti-androgens and taxanes are a mainstay of prostate cancer (PCa) treatment. These therapies, however, fail as the tumor progresses. Molecular characterization of advanced refractory PCa to identify actionable alterations was undertaken to enhance current treatment paradigms. Methods: 388 PCa cases were evaluated. A multiplatform approach of biomarker testing was used and included a combination of sequencing (NGS), protein expression (IHC) and gene amplification (ISH). Results: IHC was performed in ~350 patients, ISH was performed ~100 patients. Therapies not commonly used in PCa may be considered: cetuximab (47% EGFR IHC high, 13% EGFR amplified) and temozolomide, gemcitabine and fluorouracil based on low expression of MGMT (44%), RRM1 (69%) and TS (83%). Traditional PCa drugs including taxanes based on low TUBB3 (74%) or high TLE3 (45%), anti-androgens based on high AR (89%) and platinums based on low ERCC1 (72%) may also be of benefit. Based on NGS data from ~66 patients, the most commonly mutated pathways are TP53 (43%), PI3K/PTEN (16%) and Wnt (APC or CTNNB1) (11%); agents targeting these pathways are available in clinical trials. Single agent response rates in PCa have been disappointing or patients relapse. Additional analyses identified potential combination strategies. Forty percent of patients with low TS or RRM1 harbor mutations in TP53. Recent data (Blackwood, 2013) suggest the addition of Chk1 inhibitors potentiates response to chemotherapy in TP53-mutated patients. Dual targeting of AR and PIK3CA may be of potential benefit as 68% of patients with high AR expression exhibit PTEN loss. Twenty-one percent of patients exhibit low TUBB3 and high TLE3, a majority of which exhibit low ERCC1 (75%) or high expression of AR (98%), suggesting combination of taxanes plus carboplatin or anti-androgens, respectively. Forty-four percent of PCa lack actionable gene mutations by NGS, however 96% of these patients did have actionable targets identified by IHC and ISH platforms. Conclusions: Combining novel targeted agents with traditional treatments may achieve additive or synergistic effects. Utilization of IHC, ISH and mutational analysis results can identify combination treatment strategies for advanced refractory PCa.