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OC.04.5 EUS-FNA FOR KRAS MUTATIONAL ANALYSIS IN PANCREATIC LESIONS, PRELIMINARY RESULTS USING THREE DIFFERENT TECHNIQUES: SANGER SEQUENCING, NEXT GENERATION SEQUENCING AND REAL-TIME PCR
Abstracts of the 18th National Congress of Digestive Diseases / Digestive and Liver Disease 44S (2012) S55–S220 patient underwent esophageal EUS performed with the small diameter EBUS (Pentax EB1970UK). Results: Twenty-four patients (18M, mean age 61.5) were scheduled to have EUS evaluation for esophageal cancer staging. EUS examination was successfully in 16 patients (66.6%). In the remaining 8 patients (33.3%) the esophageal lesion precluded the passage of a standard echoendoscope. The EBUS scope was successfully passed through the stricture in all 8 cases. We were able to perform tumor staging and evaluation of lymphnodes of celiac axis and of the hepatic and splenic hilum in all cases (T2N1 in 4 patients, T3N1 in the remaining patients). Evaluation of the mediastinum, and surrounding structures was completed with excellent images. No complications were reported. Conclusions: EBUS may be a valuable tool to perform upper EUS evaluation in situations when a standard EUS scope cannot be passed because of luminal narrowing. We showed that the smaller diameter EBUS scope represents a safe and available technology to perform an accurate staging of patients with esophageal cancer.
OC.04.4 INTERVENTIONAL ENDOSCOPIC ULTRASOUND COMPLICATIONS: FINAL RESULTS OF A LARGE PROSPECTIVE MULTICENTER STUDY I. Tarantino ∗ ,1 , C. Fabbri 2 , R. Di Mitri 3 , N. Pagano 4 , N. Muscatiello 5 , L. Barresi 1 , F. Mocciaro 3 , A. Maimone 2 , M. Traina 1 1 Ismett (Istituto Mediterraneo Per I Trapianti e Le Terapie Ad Alta Specializzazione), Palermo, Italy; 2 Ospedale Bellaria-Maggiore Ausl Bologna, Bologna, Italy; 3 Arnas Ospedale Civico, Plaermo, Italy; 4 Istituto Clinico Humanitas (Rozzano), Milano, Italy; 5 Ospedale di Foggia, Foggia, Italy
Background and aim: Over the last decade, interventional endoscopic ultrasound (EUS) procedures (e.g., celiac plexus neurolysis [CPN], pseudocyst drainage, biliary drainage) have become widespread in clinical practice. However, complications related to interventional EUS have not been assessed in large prospective studies. The aim of this study was to verify the complication rates of interventional EUS and the related risk factors in a prospective, multicenter study. Material and methods: From Jan 2010 to Oct 2011, we prospectively collected data from 5 referral centers, searching for all complications related to interventional EUS. Data on demographics, comorbidities, drug histories, and laboratory tests were collected. All patients were followed-up for 2 months. Vital signs were monitored during procedures, performed under deep sedation or general anesthesia by experienced endosonographers. Antibiotic prophylaxis was administered when indicated. Early and late complications (mild, moderate, severe, fatal) were recorded during follow-up. Results: One thousand one hundred-eighteen (1118) patients (592 male), with a mean age of 63.9±14.4, underwent interventional EUS procedures: 1044 FNA (214 on cystic lesions), 43 CPN, 23 pseudocyst drainage, and 8 biliary drainage. The overall complication rate was 1.9%. Complications were: 7 fever, 4 diarrhea (in CPN), 3 cystic hemorrhages, 3 mild pancreatitis (in FNA), 2 gastric bleeding (in FNA), 1 epigastric pain (FNA), 1 desaturation. All events occurred within 5 days, and were mild in 17 patients and moderate in 4. All complications resolved with medical therapy. Rate of complications was 8.1% (6/74) after therapeutic procedure, while 1.4% (15/1044) after EUS-FNA (p<0.001, OR 6.05, 95% CI 2.27-16.09). FNA on cystic lesions was associated with a higher risk of complications than that on solid ones (5.1% versus 0.5%; p<0.001, OR 11.2, 95% CI 3.52-35.51). Conclusions: This study is the first prospective, multicenter assessment of complication rates after interventional EUS procedures. Our results confirm the overall safety of EUS-FNA. Nevertheless, the complication rate in FNA on cystic lesions is higher than on solid ones. Despite a higher complication rate associated with the therapeutic procedures compared with FNA, the risk is acceptable given the clinical setting.
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OC.04.5 EUS-FNA FOR KRAS MUTATIONAL ANALYSIS IN PANCREATIC LESIONS, PRELIMINARY RESULTS USING THREE DIFFERENT TECHNIQUES: SANGER SEQUENCING, NEXT GENERATION SEQUENCING AND REAL-TIME PCR M. Visani 1 , D. De Biase 1 , A.M. Polifemo ∗ ,2 , C. Fabbri 2 , P. Baccarini 3 , A. Maimone 2 , N. D’Imperio 2 , A. Pession 1 1 Department of Sperimental Phathology University of Bologna, Bologna, Italy; 2 Unit of Gastroenterology and Digestive Endoscopy, Ausl Bologna Bellaria-Maggiore Hospital, Bologna, Italy; 3 Unit of Anatomy and Hystophathology Bellaria Hospital, Bologna, Italy
Background and aim: KRAS is an oncogene frequently mutated in pancreatic carcinomas. 90% of the mutations are localized at the level of codons 12, 13 and 61. Mutations in KRAS are useful in discriminating benign from malignant pancreatic lesions. Identifying such mutations using FNA cytology performed under EUS preoperative endoscopy, may be an important diagnostic aid. For this reason it is necessary to have highly sensitive and specific molecular techniques. Material and methods: In thirty-five consecutive patients, who came to our observation to undergo EUS-FNA for typing of focal lesions of the pancreas, was collected cytological material for molecular analysis. The material, collected and preserved in ethyl alcohol, was used for the analysis of KRAS through: automated sequencing (capillaries, platform Beckman), Real-Time PCR (ASLNAqPCR) and Next Generation Sequencing (NGS - platform Roche-454-GS-Junior). Results: The analysis with the Beckman sequencer has identified KRAS mutations in 12/35 samples and in 2 of these double mutations were observed. Through ASLNAqPCR KRAS mutations were detected in 13/35 patients, 6 with multiple mutations. Finally, by NGS mutations were detected in 16/35 samples including 6 multiple. Conclusions: ASLNAqPCR NGS and techniques were found to be more sensitive than conventional sequencing. The NGS is able to detect infrequent mutations providing also data on the heterogeneity of the tumor. The presence of multiple mutations in individual patients could suggest the existence of multiple cell populations with different mutations (subclones) bringing, in the near future, possible implications for clinical treatment.
OC.04.6 DIAGNOSTIC VALUES OF COMBINED APPROACH OF TRANSBRONCHIAL NEEDLE ASPIRATION (TBNA) AND TRANSESOPHAGEAL ENDOSCOPIC ULTRASOUND-FINE NEEDLE ASPIRATION (EUS-FNA) IN THE DETECTION OF MEDIASTINAL MASSES D. Assisi ∗ , M. Filippetti, T. Federici, P. Visca, M. Anti Regina Elena National Cancer Institute, Rome, Italy Background and aim: Minimally invasive methods for accurate diagnosis of mediastinal masses are an advisable alternative to mediastinoscopy because of a lesser rate of complications. TBNA alone has been shown to be effective and sufficiently safe either when extensive mediastinal involvment is present or when the single anterior mediastinum has to be reached. EUS-FNA is able to collect samples from both posterior and inferior mediastinum (inaccessible to TBNA). In this study we evaluated the efficacy of a complementary approach to the morphological diagnosis of mediastinal masses through the combined use of TBNA and EUS-FNA. Material and methods: Forty-two patients with not histologically determined mediastinal masses underwent a combined technique by TBNA (videobronchoscope-Olympus) and EUS-FNA (linear echoendoscope- Olympus) under unconscious sedation. Collected samples were fixed in Thin Prep. An average of 4 needle passages were carried out during each procedure. Cytopathologic examination was performed by an experienced pathologist. When the histological report was negative for malignancy patients underwent mediastinoscopy. Results: Thirty-one out of 42 patients had a diagnosis of malignancy by TBNA/EUS-FNA combined procedure (26 primary lung cancer, 4 nodal
OC.04.4 INTERVENTIONAL ENDOSCOPIC ULTRASOUND COMPLICATIONS: FINAL RESULTS OF A LARGE PROSPECTIVE MULTICENTER STUDY I. Tarantino ∗ ,1 , C. Fabbri 2 , R. Di Mitri 3 , N. Pagano 4 , N. Muscatiello 5 , L. Barresi 1 , F. Mocciaro 3 , A. Maimone 2 , M. Traina 1 1 Ismett (Istituto Mediterraneo Per I Trapianti e Le Terapie Ad Alta Specializzazione), Palermo, Italy; 2 Ospedale Bellaria-Maggiore Ausl Bologna, Bologna, Italy; 3 Arnas Ospedale Civico, Plaermo, Italy; 4 Istituto Clinico Humanitas (Rozzano), Milano, Italy; 5 Ospedale di Foggia, Foggia, Italy
Background and aim: Over the last decade, interventional endoscopic ultrasound (EUS) procedures (e.g., celiac plexus neurolysis [CPN], pseudocyst drainage, biliary drainage) have become widespread in clinical practice. However, complications related to interventional EUS have not been assessed in large prospective studies. The aim of this study was to verify the complication rates of interventional EUS and the related risk factors in a prospective, multicenter study. Material and methods: From Jan 2010 to Oct 2011, we prospectively collected data from 5 referral centers, searching for all complications related to interventional EUS. Data on demographics, comorbidities, drug histories, and laboratory tests were collected. All patients were followed-up for 2 months. Vital signs were monitored during procedures, performed under deep sedation or general anesthesia by experienced endosonographers. Antibiotic prophylaxis was administered when indicated. Early and late complications (mild, moderate, severe, fatal) were recorded during follow-up. Results: One thousand one hundred-eighteen (1118) patients (592 male), with a mean age of 63.9±14.4, underwent interventional EUS procedures: 1044 FNA (214 on cystic lesions), 43 CPN, 23 pseudocyst drainage, and 8 biliary drainage. The overall complication rate was 1.9%. Complications were: 7 fever, 4 diarrhea (in CPN), 3 cystic hemorrhages, 3 mild pancreatitis (in FNA), 2 gastric bleeding (in FNA), 1 epigastric pain (FNA), 1 desaturation. All events occurred within 5 days, and were mild in 17 patients and moderate in 4. All complications resolved with medical therapy. Rate of complications was 8.1% (6/74) after therapeutic procedure, while 1.4% (15/1044) after EUS-FNA (p<0.001, OR 6.05, 95% CI 2.27-16.09). FNA on cystic lesions was associated with a higher risk of complications than that on solid ones (5.1% versus 0.5%; p<0.001, OR 11.2, 95% CI 3.52-35.51). Conclusions: This study is the first prospective, multicenter assessment of complication rates after interventional EUS procedures. Our results confirm the overall safety of EUS-FNA. Nevertheless, the complication rate in FNA on cystic lesions is higher than on solid ones. Despite a higher complication rate associated with the therapeutic procedures compared with FNA, the risk is acceptable given the clinical setting.
S65
OC.04.5 EUS-FNA FOR KRAS MUTATIONAL ANALYSIS IN PANCREATIC LESIONS, PRELIMINARY RESULTS USING THREE DIFFERENT TECHNIQUES: SANGER SEQUENCING, NEXT GENERATION SEQUENCING AND REAL-TIME PCR M. Visani 1 , D. De Biase 1 , A.M. Polifemo ∗ ,2 , C. Fabbri 2 , P. Baccarini 3 , A. Maimone 2 , N. D’Imperio 2 , A. Pession 1 1 Department of Sperimental Phathology University of Bologna, Bologna, Italy; 2 Unit of Gastroenterology and Digestive Endoscopy, Ausl Bologna Bellaria-Maggiore Hospital, Bologna, Italy; 3 Unit of Anatomy and Hystophathology Bellaria Hospital, Bologna, Italy
Background and aim: KRAS is an oncogene frequently mutated in pancreatic carcinomas. 90% of the mutations are localized at the level of codons 12, 13 and 61. Mutations in KRAS are useful in discriminating benign from malignant pancreatic lesions. Identifying such mutations using FNA cytology performed under EUS preoperative endoscopy, may be an important diagnostic aid. For this reason it is necessary to have highly sensitive and specific molecular techniques. Material and methods: In thirty-five consecutive patients, who came to our observation to undergo EUS-FNA for typing of focal lesions of the pancreas, was collected cytological material for molecular analysis. The material, collected and preserved in ethyl alcohol, was used for the analysis of KRAS through: automated sequencing (capillaries, platform Beckman), Real-Time PCR (ASLNAqPCR) and Next Generation Sequencing (NGS - platform Roche-454-GS-Junior). Results: The analysis with the Beckman sequencer has identified KRAS mutations in 12/35 samples and in 2 of these double mutations were observed. Through ASLNAqPCR KRAS mutations were detected in 13/35 patients, 6 with multiple mutations. Finally, by NGS mutations were detected in 16/35 samples including 6 multiple. Conclusions: ASLNAqPCR NGS and techniques were found to be more sensitive than conventional sequencing. The NGS is able to detect infrequent mutations providing also data on the heterogeneity of the tumor. The presence of multiple mutations in individual patients could suggest the existence of multiple cell populations with different mutations (subclones) bringing, in the near future, possible implications for clinical treatment.
OC.04.6 DIAGNOSTIC VALUES OF COMBINED APPROACH OF TRANSBRONCHIAL NEEDLE ASPIRATION (TBNA) AND TRANSESOPHAGEAL ENDOSCOPIC ULTRASOUND-FINE NEEDLE ASPIRATION (EUS-FNA) IN THE DETECTION OF MEDIASTINAL MASSES D. Assisi ∗ , M. Filippetti, T. Federici, P. Visca, M. Anti Regina Elena National Cancer Institute, Rome, Italy Background and aim: Minimally invasive methods for accurate diagnosis of mediastinal masses are an advisable alternative to mediastinoscopy because of a lesser rate of complications. TBNA alone has been shown to be effective and sufficiently safe either when extensive mediastinal involvment is present or when the single anterior mediastinum has to be reached. EUS-FNA is able to collect samples from both posterior and inferior mediastinum (inaccessible to TBNA). In this study we evaluated the efficacy of a complementary approach to the morphological diagnosis of mediastinal masses through the combined use of TBNA and EUS-FNA. Material and methods: Forty-two patients with not histologically determined mediastinal masses underwent a combined technique by TBNA (videobronchoscope-Olympus) and EUS-FNA (linear echoendoscope- Olympus) under unconscious sedation. Collected samples were fixed in Thin Prep. An average of 4 needle passages were carried out during each procedure. Cytopathologic examination was performed by an experienced pathologist. When the histological report was negative for malignancy patients underwent mediastinoscopy. Results: Thirty-one out of 42 patients had a diagnosis of malignancy by TBNA/EUS-FNA combined procedure (26 primary lung cancer, 4 nodal