Observational Study
Copyright ©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. May 14, 2023; 29(18): 2864-2874
Published online May 14, 2023. doi: 10.3748/wjg.v29.i18.2864
Endoscopic ultrasound-guided fine-needle aspiration pancreatic adenocarcinoma samples yield adequate DNA for next-generation sequencing: A cohort analysis
Stefania Bunduc, Bianca Varzaru, Razvan Andrei Iacob, Andrei Sorop, Ioana Manea, Andreea Spiridon, Raluca Chelaru, Adina Emilia Croitoru, Gabriel Becheanu, Mona Dumbrava, Simona Dima, Irinel Popescu, Cristian Gheorghe
Stefania Bunduc, Bianca Varzaru, Razvan Andrei Iacob, Gabriel Becheanu, Simona Dima, Cristian Gheorghe, Faculty of Medicine, Carol Davila University of Medicine and Pharmacy, Bucharest 020021, Romania
Stefania Bunduc, Razvan Andrei Iacob, Adina Emilia Croitoru, Gabriel Becheanu, Mona Dumbrava, Simona Dima, Irinel Popescu, Cristian Gheorghe, Digestive Diseases and Liver Transplantation Center, Fundeni Clinical Institute, Bucharest 022328, Romania
Stefania Bunduc, Bianca Varzaru, Razvan Andrei Iacob, Andrei Sorop, Ioana Manea, Andreea Spiridon, Raluca Chelaru, Adina Emilia Croitoru, Gabriel Becheanu, Mona Dumbrava, Simona Dima, Irinel Popescu, Cristian Gheorghe, Center of Excellence in Translational Medicine, Fundeni Clinical Institute, Bucharest 022328, Romania
Author contributions: Bunduc S contributed to conceptualization, methodology, investigation, formal analysis, original draft, and data visualization; Varzaru B contributed to conceptualization, investigation, data curation, methodology, review and editing; Iacob RA contributed to conceptualization, supervision, review and editing, methodology, funding acquisition, and resources; Sorop A contributed to methodology, investigation, data curation, review and editing; Manea I contributed to methodology, investigation, data curation, review and editing; Spiridon A contributed to investigation, data curation, review and editing; Chelaru R contributed to investigation, data curation, review and editing; Becheanu G contributed to conceptualization, methodology, investigation, review and editing; Dumbrava M contributed to conceptualization, methodology, investigation, review and editing; Croitoru A contributed to conceptualization, methodology, review and editing, Dima S contributed to conceptualization, supervision, funding acquisition, project administration, review and editing, resources; Popescu I contributed to conceptualization, supervision, funding acquisition, project administration, review and editing, Gheorghe C contributed to guarantor of the article, conceptualization, supervision, funding acquisition, project administration, review and editing; all authors certify that they have participated sufficiently in this work to take public responsibility for the content, including participation in the concept, design, analysis, writing, or revision of the manuscript. All authors have read the journal's authorship agreement, and reviewed and approved the final version of the manuscript.
Supported by The Executive Agency for Higher Education, Research, Development and Innovation Funding-research, No. PN-III-P1-1.2-PCCDI-2017-0797 (PANCNGS).
Institutional review board statement: This prospective observational study was approved by the Internal Review Board of Fundeni Clinical Institute, No. IRB:23878/14.06.2018.
Informed consent statement: Each participant gave informed consent for study enrolment.
Conflict-of-interest statement: All authors confirm they have no conflict of interest in relation to this manuscript.
Data sharing statement: The data underlying this article will be shared upon reasonable request to the corresponding author.
STROBE statement: The authors have read the STROBE Statement-checklist of items, and the manuscript was prepared and revised according to the STROBE Statement-checklist of items.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Cristian Gheorghe, FASGE, MD, PhD, Professor, Faculty of Medicine, Carol Davila University of Medicine and Pharmacy, No. 258 Fundeni Road, Bucharest 020021, Romania. drcgheorghe@gmail.com
Received: December 31, 2022
Peer-review started: December 31, 2022
First decision: February 28, 2023
Revised: March 14, 2023
Accepted: April 18, 2023
Article in press: April 18, 2023
Published online: May 14, 2023
Processing time: 130 Days and 10.9 Hours
ARTICLE HIGHLIGHTS
Research background

Due to the opportunities for personalized treatment in pancreatic adenocarcinoma (PDAC), genetic testing is increasingly performed. Fine needle biopsy is the method recommended for endoscopic ultrasound (EUS) guided tissue acquisition to obtain samples dedicated to downstream comprehensive molecular analyses. In current practice however, fine needle aspiration (FNA) is more widely accessible.

Research motivation

We evaluated the EUS-FNA PDAC samples in terms of adequacy for next generation sequencing (NGS) of the yielded DNA to assess the possibility of using this type of samples for genetic testing.

Research objectives

To investigate the association between DNA parameters (amount and purity) measured by spectrophotometry and FNA needle size (19 gauge [G] or 22G), and also tumor characteristics.

Research methods

We performed an observational prospective study on PDAC cases diagnosed through EUS-FNA at a tertiary center of Gastroenterology in Romania. During EUS one pass acquired samples dedicated to genetic testing. NGS adequacy was a dichotomus variable defined based on DNA parameters (purity: A260/280 ≥ 1.7 and DNA amount: ≥ 100 ng for whole genome sequencing, ≥ 50 ng for whole exome sequencing or ≥ 10 ng for amplicon based targeted NGS).

Research results

Our cohort analysis comprised 105 confirmed PDAC cases. The majority of samples were acquired with 22G FNA needles-75 (71%). The DNA amount was in average 1289 ng (inter-quartile range: 534.75-2995). All samples yielded more than 10 ng of DNA while 98 (93%) of them yielded more than 100 ng of DNA. Needle size was not correlated with DNA NGS adequacy rate regardless of NGS type. Needle size did not influence the concentration or A260/280 ratio of the extracted DNA. The median A260/230 was significantly higher in the 22G samples than in 19G samples (P = 0.038). In multivariate analysis (on needle size, tumor location and tumor diameter) the only independent predictor of A260/230 was needle size (β = 0.36, t(104) = 2.1, P = 0.038).

Research conclusions

Both 22G an 19G EUS-FNA PDAC samples are adequate for downstream NGS. FNA needle size and tumor characteristics did not significantly influence sample NGS adequacy rate. Greater FNA needle size might be associated with decreased sample purity.

Research perspectives

PDAC FNA samples (22 and 19G) yield samples of adequate purity and amount for NGS and can be used both in current practice and for research purposes.