title: Targeted next generation sequencing of endoscopic ultrasound acquired cytology from ampullary and pancreatic adenocarcinoma has the potential to aid patient stratification for optimal therapy selection.
abstract: OBJECTIVE
  Less than 10% of registered drug intervention trials for pancreatic ductal adenocarcinoma (PDAC) include a biomarker stratification strategy. The ability to identify distinct mutation subsets via endoscopic ultrasound fine needle aspiration (EUS FNA) molecular cytology could greatly aid clinical trial patient stratification and offer predictive markers. We identified chemotherapy treatment naïve ampullary adenocarcinoma and PDAC patients who underwent EUS FNA to assess multigene mutational frequency and diversity with a surgical resection concordance assessment, where available.
  
  
  METHODS
  Following strict cytology smear screening criteria, targeted next generation sequencing (NGS) using a 160 cancer gene panel was performed.
  
  
  RESULTS
  Complete sequencing was achieved in 29 patients, whereby 83 pathogenic alterations were identified in 21 genes. Cytology genotyping revealed that the majority of mutations were identified in KRAS (93%), TP53 (72%), SMAD4 (31%), and GNAS (10%). There was 100% concordance for the following pathogenic alterations: KRAS, TP53, SMAD4, KMT2D, NOTCH2, MSH2, RB1, SMARCA4, PPP2R1A, PIK3R1, SCL7A8, ATM, and FANCD2. Absolute multigene mutational concordance was 83%. Incremental cytology smear mutations in GRIN2A, GATA3 and KDM6A were identified despite re-examination of raw sequence reads in the corresponding resection specimens.
  
  
  CONCLUSIONS
  EUS FNA cytology genotyping using a 160 cancer gene NGS panel revealed a broad spectrum of pathogenic alterations. The fidelity of cytology genotyping to that of paired surgical resection specimens suggests that EUS FNA represents a suitable surrogate and may complement the conventional stratification criteria in decision making for therapies and may guide future biomarker driven therapeutic development.
authors.affiliation: Division of Gastroenterology & Hepatology, Mayo Clinic Rochester, MN, USA.
authors.forename: Ferga C
authors.initials: FC
authors.lastname: Gleeson
authors.affiliation: Department of Laboratory Medicine & Pathology, Mayo Clinic Rochester, MN, USA.
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Kerr
authors.affiliation: Department of Laboratory Medicine & Pathology, Mayo Clinic Rochester, MN, USA.
authors.forename: Benjamin R
authors.initials: BR
authors.lastname: Kipp
authors.affiliation: Department of Laboratory Medicine & Pathology, Mayo Clinic Rochester, MN, USA.
authors.forename: Jesse S
authors.initials: JS
authors.lastname: Voss
authors.affiliation: Department of Laboratory Medicine & Pathology, Mayo Clinic Rochester, MN, USA.
authors.forename: Douglas M
authors.initials: DM
authors.lastname: Minot
authors.affiliation: Division of Biomedical Statics & Informatics, Department of Health Sciences Research, Mayo Clinic Rochester, MN, USA.
authors.forename: Zheng Jin
authors.initials: ZJ
authors.lastname: Tu
authors.affiliation: Department of Laboratory Medicine & Pathology, Mayo Clinic Rochester, MN, USA.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Henry
authors.affiliation: Department of Laboratory Medicine & Pathology, Mayo Clinic Rochester, MN, USA.
authors.forename: Rondell P
authors.initials: RP
authors.lastname: Graham
authors.affiliation: Center for Individualized Medicine, Mayo Clinic, Rochester, MN, USA.
authors.forename: George
authors.initials: G
authors.lastname: Vasmatzis
authors.affiliation: Center for Individualized Medicine, Mayo Clinic, Rochester, MN, USA.
authors.forename: John C
authors.initials: JC
authors.lastname: Cheville
authors.affiliation: Division of Gastroenterology & Hepatology, Mayo Clinic Rochester, MN, USA.|Center for Individualized Medicine, Mayo Clinic, Rochester, MN, USA.
authors.forename: Konstantinos N
authors.initials: KN
authors.lastname: Lazaridis
authors.affiliation: Division of Gastroenterology & Hepatology, Mayo Clinic Rochester, MN, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Levy
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000473:pathology* / Q000628:therapy; D000368:Aged; D000369:Aged, 80 and over; D014670:Ampulla of Vater; D003138:Common Bile Duct Neoplasms / Q000235:genetics / Q000473:pathology* / Q000628:therapy; D061765:Endoscopic Ultrasound-Guided Fine Needle Aspiration / Q000379:methods*; D005260:Female; D016158:Genes, p53; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000473:pathology* / Q000628:therapy; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics; D051901:Smad4 Protein / Q000235:genetics
chemical_list: C117307:KRAS protein, human; C497796:SMAD4 protein, human; D051901:Smad4 Protein; D016283:Proto-Oncogene Proteins p21(ras)
keywords: endoscopic ultrasound fine needle aspiration; mutation concordance; pancreatic adenocarcinoma; personalized medicine; targeted next-generation sequencing