Silvia Carrara1, Giulia Soldà2, Milena Di Leo1, Daoud Rahal3, Clelia Peano4, Michele Giunta2, Laura Lamonaca1, Francesco Auriemma5, Andrea Anderloni1, Alessandro Fugazza1, Roberta Maselli1, Alberto Malesci6, Luigi Laghi7, Alessandro Repici8. 1. Department of Gastroenterology, Endoscopic Unit, Humanitas Clinical and Research Center-IRCCS, Rozzano, Italy. 2. Department of Biomedical Sciences, Humanitas University, Milan, Italy; Biology Department, Humanitas Clinical and Research Center- IRCCS, Rozzano, Italy. 3. Department of Pathology, Humanitas Clinical and Research Center-IRCCS, Rozzano, Italy. 4. Genomic Unit, Humanitas Clinical and Research Center-IRCCS, Rozzano, Italy; Institute of Genetic and Biomedical Research, UoS Milan, National Research Council, Rozzano, Italy. 5. Endoscopic Unit, Humanitas Mater Domini, Castellanza, Varese, Italy. 6. Department of Gastroenterology, Humanitas Clinical and Research Center-IRCCS, Rozzano, Italy. 7. Department of Gastroenterology, Humanitas Clinical and Research Center-IRCCS, Rozzano, Italy; Hereditary Cancer Genetics Clinic, Humanitas Clinical and Research Center-IRCCS, Rozzano, Italy; Laboratory of Molecular Gastroenterology, Humanitas Clinical and Research Center-IRCCS, Rozzano, Italy. 8. Department of Gastroenterology, Endoscopic Unit, Humanitas Clinical and Research Center-IRCCS, Rozzano, Italy; Department of Biomedical Sciences, Humanitas University, Milan, Italy.
Abstract
BACKGROUND AND AIMS: EUS-guided biopsy sampling is the method of choice for obtaining pancreatic tissue. Next-generation sequencing (NGS) has been applied to EUS-guided biopsy sampling and may classify patients based on specific molecular profiles. Our study aimed to compare side-by-side the diagnostic yield achievable by genetic identification of somatic mutations detected with NGS versus histologic and cytologic typing in locally advanced pancreatic carcinoma (LAPC) in samples acquired under EUS guidance. METHODS: We conducted a prospective comparative pilot study at Humanitas Research Hospital. The study included 33 patients referred for LAPC who underwent EUS-guided tissue acquisition using a 22-gauge Franseen needle. Material was obtained for both pathologic diagnosis and DNA extraction and targeted NGS analysis with the AmpliSeq Comprehensive Panel v3 (Illumina Inc, San Diego, Calif, USA). Twenty-one genes were prioritized for somatic mutation detection. RESULTS: The final diagnosis was pancreatic ductal adenocarcinoma (PDAC) in all patients (100%). A macroscopic core was obtained in 30 patients (91%). In 3 lesions no cores adequate for histologic analysis were obtained, but cytologic analysis revealed tumoral cells from PDAC. DNA was extracted from 32 of 33 samples (97%), most of which (27/32) carried at least 2 clearly pathogenic mutations in different genes. Detection of K-ras mutation allowed for molecular diagnosis of PDAC in most of the patients (30/32). CONCLUSIONS: In our study we demonstrated that proper tissue specimens obtained under EUS guidance allowed DNA sample extraction and subsequent NGS analysis in 97% of cases. These results support the potential role of NGS as a complementary diagnostic test to be implemented in association with standard diagnostic modalities. (Clinical trial registration number: NCT03578939.).
BACKGROUND AND AIMS: EUS-guided biopsy sampling is the method of choice for obtaining pancreatic tissue. Next-generation sequencing (NGS) has been applied to EUS-guided biopsy sampling and may classify patients based on specific molecular profiles. Our study aimed to compare side-by-side the diagnostic yield achievable by genetic identification of somatic mutations detected with NGS versus histologic and cytologic typing in locally advanced pancreatic carcinoma (LAPC) in samples acquired under EUS guidance. METHODS: We conducted a prospective comparative pilot study at Humanitas Research Hospital. The study included 33 patients referred for LAPC who underwent EUS-guided tissue acquisition using a 22-gauge Franseen needle. Material was obtained for both pathologic diagnosis and DNA extraction and targeted NGS analysis with the AmpliSeq Comprehensive Panel v3 (Illumina Inc, San Diego, Calif, USA). Twenty-one genes were prioritized for somatic mutation detection. RESULTS: The final diagnosis was pancreatic ductal adenocarcinoma (PDAC) in all patients (100%). A macroscopic core was obtained in 30 patients (91%). In 3 lesions no cores adequate for histologic analysis were obtained, but cytologic analysis revealed tumoral cells from PDAC. DNA was extracted from 32 of 33 samples (97%), most of which (27/32) carried at least 2 clearly pathogenic mutations in different genes. Detection of K-ras mutation allowed for molecular diagnosis of PDAC in most of the patients (30/32). CONCLUSIONS: In our study we demonstrated that proper tissue specimens obtained under EUS guidance allowed DNA sample extraction and subsequent NGS analysis in 97% of cases. These results support the potential role of NGS as a complementary diagnostic test to be implemented in association with standard diagnostic modalities. (Clinical trial registration number: NCT03578939.).