原句：Cancer of unknown primary remains a challenging clinical entity. Despite receiving empiric chemotherapy, median overall survival is approximately 6-12 months. Site-specific therapy based on molecular characterization has been shown to improve outcomes; however, feasibility outside of clinical trials, especially in community centers, is lacking. This study explores the application of rapid next-generation sequencing in defining cancer of unknown primary and to identify therapeutic biomarkers. 来源：这项研究探讨了快速下一代测序技术在定义原发癌未知的肿瘤（Cancer of unknown primary，CUP）和鉴定治疗生物标志物方面的应用。尽管接受了经验证的化疗，CUP的中位总生存期约为6-12个月，这仍然是一个具有挑战性的临床实体。基于分子特征的部位特异性治疗已被证明可以改善预后，但在临床试验以外，特别是在社区中心的可行性仍然不足。 翻译：原发癌未知的肿瘤仍然是具有挑战性的临床实体。尽管接受了经验证的化疗，中位总生存期约为6-12个月。基于分子特征的部位特异性治疗已被证明可以改善预后，但在临床试验以外，特别是在社区中心的可行性仍然不足。本研究探讨了快速下一代测序技术在定义原发癌未知的肿瘤和鉴定治疗生物标志物方面的应用。

Cancer of unknown primary remains a challenging clinical entity. Despite receiving empiric chemotherapy, median overall survival is approximately 6-12 months. Site-specific therapy based on molecular characterization has been shown to improve outcomes; however, feasibility outside of clinical trials, especially in community centers, is lacking. This study explores the application of rapid next-generation sequencing in defining cancer of unknown primary and to identify therapeutic biomarkers.A retrospective chart review was performed by identifying pathological samples designated cancer of unknown primary. Next-generation sequencing testing was based on an automated workflow utilizing the Genexus integrated sequencer, validated for clinical use. Genomic profiling was further integrated within a routine immunohistochemistry service, with results reported directly by anatomic pathologists.Between October 2020 and October 2021, 578 solid tumor samples underwent genomic profiling. Among this cohort, 40 were selected based on an initial diagnosis of cancer of unknown primary. The median (range) age at diagnosis was 70 (42-85) and 23 (57%) were female. Genomic data were used to support a site-specific diagnosis in 6 patients (15%). Median turnaround time was 3 business days (IQR: 1-5). Most common alterations identified were KRAS (35%), CDKN2A (15%), TP53 (15%), and ERBB2 (12%). Actionable molecular targeted therapies were identified in 23 (57%) patients, including alterations in BRAF, CDKN2A, ERBB2, FGFR2, IDH1, and KRAS. Immunotherapy-sensitizing mismatch repair deficiency was identified in 1 patient.This study supports the adoption of rapid next-generation sequencing among patients with cancer of unknown primary. We also demonstrate the feasibility of integration of genomic profiling with diagnostic histopathology and immunohistochemistry in a community practice setting. Diagnostic algorithms incorporating genomic profiling to better define cancer of unknown primary should be considered for future study.© The Author(s) 2023. Published by Oxford University Press.