Background: CC has poor prognosis and limited therapeutic options beyond first-line therapy. It is molecularly heterogeneous with several gene alterations (alt) that can be matched to targeted treatments in CT. We investigated the impact of CC molecular profiling in the clinics. Methods: From 2011 to 2017, we identified 165 patients (pts) with advanced CC - 129 intrahepatic CC (ICC) and 36 extrahepatic CC (ECC) - whose tumors were analyzed in our center with NGS tests (124 had fusion panels). We retrospectively collected outcome information and access to CT. Results: Most pts were diagnosed at stage IV (67%) and received first-line gemcitabine plus platinum (80%). Most common alt found in ICC were mutations (mt) in TP53 (19%), IDH1 (14%), CDKN2A (9%), IDH2 (7%), ATM (7%), BAP1 (6%), PIK3CA (6%), KRAS (6%), NRAS (5%), BRAF (4%) and fusions in FGFR2/3 genes (3%). In ECC, we found mt in TP53 (28%), CDKN2A (11%), PTEN (11%), PIK3CA (6%), BRCA2 (6%) and mt or amplifications (amp) in ERBB2 (11%). With a median follow up of 56 months (m), median overall survival (OS) of the overall population was 23 m, with no differences between ICC and ECC (p = 0.83). There was statistically significant difference in OS between pts with TP53 mt vs TP53 wild type (wt) tumors (28 m vs 15 m, HR 1.8, p = 0.01). During first-line therapy, median progression free survival (PFS) was 6.6 m in TP53 mt and 3.8 m in TP53 wt patients (HR 2.3, p < 0.01). 40 pts (24%) were included in a CT, with 24 pts (15%) being matched by molecular alt. Most frequent matches were FGFR inhibitor (inh) (25% of pts with targetable fusion), IDH inh in 5 (21%), PI3K inh in 3 (13%), BRAF inh in 3 (13%). Among pts treated in matched CT, median PFS was 4 m; 2 pts (8%) achieved a partial response (1 MET amp;1 BRAF mut) and 11 (46%) had stable disease as best response. We found no associations between targetable gene alt and OS or PFS during first-line therapy. Conclusions: CC is a heterogeneous tumor with a broad spectrum of molecular alt that have prognostic and treatment implications. Matched therapies may confer clinical benefit. Molecular profiling of CC is of growing interest to improve the knowledge of this disease and its therapeutic opportunities.