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©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.
Exosomal glypican-1 is elevated in pancreatic cancer precursors and can signal genetic predisposition in the absence of endoscopic ultrasound abnormalities
Pedro Moutinho-Ribeiro, Ines A Batista, Sofia T Quintas, Bárbara Adem, Marco Silva, Rui Morais, Armando Peixoto, Rosa Coelho, Pedro Costa-Moreira, Renato Medas, Susana Lopes, Filipe Vilas-Boas, Manuela Baptista, Diogo Dias-Silva, Ana L Esteves, Filipa Martins, Joanne Lopes, Helena Barroca, Fátima Carneiro, Guilherme Macedo, Sonia A Melo
Pedro Moutinho-Ribeiro, Marco Silva, Rui Morais, Armando Peixoto, Rosa Coelho, Pedro Costa-Moreira, Renato Medas, Susana Lopes, Filipe Vilas-Boas, Guilherme Macedo, Serviço de Gastrenterologia, Centro Hospitalar Universitário de São João, Porto 4200, Portugal
Pedro Moutinho-Ribeiro, Marco Silva, Rui Morais, Armando Peixoto, Rosa Coelho, Pedro Costa-Moreira, Renato Medas, Susana Lopes, Filipe Vilas-Boas, Fátima Carneiro, Guilherme Macedo, Sonia A Melo, Faculty of Medicine, University of Porto, Porto 4200, Portugal
Ines A Batista, Sofia T Quintas, Bárbara Adem, Fátima Carneiro, Sonia A Melo, Instituto de Investigação e Inovação em Saúde (i3S), Universidade do Porto, Porto 4200, Portugal
Ines A Batista, Sofia T Quintas, Fátima Carneiro, Sonia A Melo, IPATIMUP–Institute of Molecular Pathology and Immunology, University of Porto, Porto 4200, Portugal
Ines A Batista, Bárbara Adem, Instituto de Ciências Biomédicas Abel Salazar (ICBAS), University of Porto, Porto 4050, Portugal
Manuela Baptista, Serviço de Cirurgia Geral, Centro Hospitalar Universitário de São João, Porto 4200, Portugal
Diogo Dias-Silva, Ana L Esteves, Filipa Martins, Unidade de Saúde Familiar Serpa Pinto, ACeS Porto Ocidental, Porto 4250, Portugal
Joanne Lopes, Helena Barroca, Fátima Carneiro, Serviço de Anatomia Patológica, Centro Hospitalar Universitário de São João, Porto 4200, Portugal
Author contributions: Moutinho-Ribeiro P was the main project investigator, was responsible for patient selection and sample collection, performed pancreatic endoscopic ultrasound, and wrote the initial manuscript; Batista IA participated in the sample processing, supervised and collected the data, performed the data analyses, and supported the manuscript writing; Quintas ST participated in the sample processing, data collection and analyses, assembled the figures, and supported the manuscript writing; Adem B participated in the sample processing and supported the data analyses; Silva M participated in the data collection and performed the statistical analyses; Morais R, Peixoto A, Coelho R, Costa-Moreira P, and Medas R participated in patient observation, data collection, and database management; Medas R performed the statistical analyses; Lopes S and Vilas-Boas F performed the pancreatic endoscopic ultrasound; Baptista M, Dias-Silva D, Esteves AL, and Martins F participated in patient observation and data collection; Lopes J, Barroca H, and Carneiro F performed the cytological and histological examination of the pancreatic specimens; Macedo G conceived and designed the study, interpreted the data analyses, and critically reviewed the manuscript; Melo SA conceived and designed the study, supervised the data analyses, and critically reviewed the manuscript; All authors approved the final version to be published.
Supported by Guilherme Macedo team was supported by the Portuguese Society of Digestive Endoscopy (SPED) 2017 Research Grant, No. SG/CHSJ-A2017; Norte Portugal Regional Programme (NORTE 2020) under the PORTUGAL 2020 Partnership Agreement through the European Regional Development Fund (ERDF) to Sonia A Melo, No. NORTE-01-0145-FEDER-000029; National Funds through Foundation for Science and Technology (FCT) to Sonia A Melo, No. POCI-01-0145-FEDER-32189; and Foundation for Science and Technology (FCT) to Bárbara Adem and Ines A Batista, No. PD/BD/135546/2018 and No. SFRH/BD/144854/2019.
Institutional review board statement: This study was reviewed and approved by the Ethics Committee of Centro Hospitalar Universitário de São João (CHUSJ), Porto, Portugal, No. CES 327-15.
Informed consent statement: All study participants or their legal guardian provided informed written consent about personal and medical data collection prior to study enrolment.
Conflict-of-interest statement: Sónia A Melo holds patents in the field of exosomes biology and are licensed to Codiak Biosciences, Inc. All other authors have no conflicts of interest to declare.
Data sharing statement: No additional data are available.
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: Sonia A Melo, PhD, Academic Research, Assistant Professor, Instituto de Investigação e Inovação em Saúde (i3S), Universidade do Porto, R. Alfredo Allen 208, Porto 4200, Portugal.
smelo@i3s.up.pt
Received: February 17, 2022
Peer-review started: February 17, 2022
First decision: April 16, 2022
Revised: April 30, 2022
Accepted: June 24, 2022
Article in press: June 24, 2022
Published online: August 21, 2022
Processing time: 179 Days and 20.5 Hours
BACKGROUND
Individuals within specific risk groups for pancreatic ductal adenocarcinoma (PDAC) [mucinous cystic lesions (MCLs), hereditary risk (HR), and new-late onset diabetes mellitus (NLOD)] represent an opportunity for early cancer detection. Endoscopic ultrasound (EUS) is a premium image modality for PDAC screening and precursor lesion characterization. While no specific biomarker is currently clinically available for this purpose, glypican-1 (GPC1) is overexpressed in the circulating exosomes (crExos) of patients with PDAC compared with healthy subjects or those harboring benign pancreatic diseases.
AIM
To evaluate the capacity of GPC1+ crExos to identify individuals at higher risk within these specific groups, all characterized by EUS.
METHODS
This cross-sectional study with a prospective unicentric cohort included 88 subjects: 40 patients with MCL, 20 individuals with HR, and 20 patients with NLOD. A control group (CG) was submitted to EUS for other reasons than pancreatic pathology, with normal pancreas and absence of hereditary risk factors (n = 8). The inclusion period was between October 2016 and January 2019, and the study was approved by the Ethics Committee of Centro Hospitalar Universitário de São João, Porto, Portugal. All patients provided written informed consent. EUS and blood tests for quantification of GPC1+ crExos by flow cytometry and carbohydrate antigen 19-9 (CA 19-9) levels by ELISA were performed in all subjects. EUS-guided tissue acquisition was done whenever necessary. For statistical analysis, SPSS® 27.0 (IBM Corp., Armonk, NY, United States) version was used. All graphs were created using GraphPad Prism 7.00 (GraphPad Software, San Diego, CA, United States).
RESULTS
Half of MCLs harbored worrisome features (WF) or high-risk stigmata (HRS). Pancreatic abnormalities were detected by EUS in 10.0% and 35.0% in HR and NLOD individuals, respectively, all considered non-malignant and “harmless.” Median levels of GPC1+ crExos were statistically different: MCL [99.4%, interquartile range (IQR): 94.9%-99.8%], HR (82.0%, IQR: 28.9%-98.2%), NLOD (12.6%, IQR: 5.2%-63.4%), and CG (16.2%, IQR: 6.6%-20.1%) (P < 0.0001). Median levels of CA 19-9 were within the normal range in all groups (standard clinical cut-off of 37 U/mL). Within HR, individuals with a positive history of cancer had higher median levels of GPC1+ crExos (97.9%; IQR: 61.7%-99.5%), compared to those without (59.7%; IQR: 26.3%-96.4%), despite no statistical significance (P = 0.21). Pancreatic cysts with WF/HRS were statistically associated with higher median levels of GPC1+ crExos (99.6%; IQR: 97.6%-99.8%) compared to those without (96.5%; IQR: 81.3%-99.5%) (P = 0.011), presenting an area under the receiver operating characteristic curve value of 0.723 (sensitivity 75.0% and specificity 67.7%, using a cut-off of 98.5%; P = 0.012).
CONCLUSION
GPC1+ crExos may act as biomarker to support the diagnosis and stratification of PDAC precursor lesions, and in signaling individuals with genetic predisposition in the absence of EUS abnormalities.
Core Tip: Patients with mucinous cystic lesions (MCLs), hereditary risk (HR), and new-late onset diabetes mellitus represent a target for early pancreatic ductal adenocarcinoma (PDAC) detection. Within these groups, we evaluated the capacity of glypican-1 positive (GPC1+) circulating exosomes (crExos) to identify individuals at higher risk, all characterized by endoscopic ultrasound (EUS). High levels of GPC1+ crExos were present in MCL and in individuals with HR (predominantly in those with history of cancer), even in the absence of EUS abnormalities. GPC1 may represent a biomarker to support the diagnosis and stratification of PDAC precursor lesions, and indicate genetic predisposition.
