Javier Bosque1, Carlos Guirao1, Asia Ferrández1, Noelia Suarez1, Maria Isabel Castillejo2,3, Diana Anguita1, María Pamies1, Alejandro Moya4, José Luis Soto2,3, Javier Gallego Plazas5. 1. Medical Oncology Department, Elche University Hospital, Camí Làlmazara s/n, 03203, Elche, Spain. 2. Molecular Genetics Unit, Elche University Hospital, 03203, Elche, Spain. 3. Foundation for the Promotion of Health and Biomedical Research of Valencia Region (FISABIO), FISABIO-Elche Health Department, 03203, Elche, Spain. 4. Biostatistics Department, FISABIO-Elche Health Department, 03203, Elche, Spain. 5. Medical Oncology Department, Elche University Hospital, Camí Làlmazara s/n, 03203, Elche, Spain. j.gallegoplazas@gmail.com.
Abstract
BACKGROUND: Cell-free DNA analysis (cfDNA) holds promise for residual disease or tumor burden quantification in colorectal cancer, with reduced costs and diagnostic equipment compared to gold standard-specific tumor DNA (ctDNA) analysis. METHODS: This prospective case-control study included 46 colorectal cancer patients and healthy controls to perform cfDNA quantification by fluorometry using Quantus Fluorometer (Promega, Madison, WI) and using cell-free DNA ScreenTape assay (Agilent) and 4200 TapeStation instrument (Agilent Technologies, Inc., Santa Clara, CA, USA). cfDNA quantification results were correlated with stage, clinical and histopathological features. RESULTS: 33 localized (8 stage I, 12 stage II, and 13 stage III) and 13 advanced colorectal cancer patients were included. No differences in cfDNA quantification by fluorometry were demonstrated depending on stage or histopathological features in localized disease patients. Differences in cfDNA quantification by fluorometry could be demonstrated in patients with advanced disease depending on the presence of liver metastases and synchronous or metachronous metastatic disease. Differences in cfDNA quantification by fluorometry could be demonstrated between advanced colorectal cancer patients and both localized disease patients and healthy controls. Secondary cfDNA analysis by electrophoresis, although showing more specificity to measure ctDNA in cfDNA values, could not improve the capacity to detect differences between analyzed a groups beyond previously achieved with fluorometry. CONCLUSION: This exploratory analysis of cfDNA based on fluorometry and electrophoresis methods showed promising results discriminating colorectal cancer and non-cancer patients, as well as different colorectal cancer stages and disease profiles. Further studies are needed to increase our knowledge and to help to overcome barriers to broader implementation and applications.
BACKGROUND: Cell-free DNA analysis (cfDNA) holds promise for residual disease or tumor burden quantification in colorectal cancer, with reduced costs and diagnostic equipment compared to gold standard-specific tumor DNA (ctDNA) analysis. METHODS: This prospective case-control study included 46 colorectal cancer patients and healthy controls to perform cfDNA quantification by fluorometry using Quantus Fluorometer (Promega, Madison, WI) and using cell-free DNA ScreenTape assay (Agilent) and 4200 TapeStation instrument (Agilent Technologies, Inc., Santa Clara, CA, USA). cfDNA quantification results were correlated with stage, clinical and histopathological features. RESULTS: 33 localized (8 stage I, 12 stage II, and 13 stage III) and 13 advanced colorectal cancer patients were included. No differences in cfDNA quantification by fluorometry were demonstrated depending on stage or histopathological features in localized disease patients. Differences in cfDNA quantification by fluorometry could be demonstrated in patients with advanced disease depending on the presence of liver metastases and synchronous or metachronous metastatic disease. Differences in cfDNA quantification by fluorometry could be demonstrated between advanced colorectal cancer patients and both localized disease patients and healthy controls. Secondary cfDNA analysis by electrophoresis, although showing more specificity to measure ctDNA in cfDNA values, could not improve the capacity to detect differences between analyzed a groups beyond previously achieved with fluorometry. CONCLUSION: This exploratory analysis of cfDNA based on fluorometry and electrophoresis methods showed promising results discriminating colorectal cancer and non-cancer patients, as well as different colorectal cancer stages and disease profiles. Further studies are needed to increase our knowledge and to help to overcome barriers to broader implementation and applications.
Authors: Florent Mouliere; Dineika Chandrananda; Anna M Piskorz; Elizabeth K Moore; James Morris; Lise Barlebo Ahlborn; Richard Mair; Teodora Goranova; Francesco Marass; Katrin Heider; Jonathan C M Wan; Anna Supernat; Irena Hudecova; Ioannis Gounaris; Susana Ros; Mercedes Jimenez-Linan; Javier Garcia-Corbacho; Keval Patel; Olga Østrup; Suzanne Murphy; Matthew D Eldridge; Davina Gale; Grant D Stewart; Johanna Burge; Wendy N Cooper; Michiel S van der Heijden; Charles E Massie; Colin Watts; Pippa Corrie; Simon Pacey; Kevin M Brindle; Richard D Baird; Morten Mau-Sørensen; Christine A Parkinson; Christopher G Smith; James D Brenton; Nitzan Rosenfeld Journal: Sci Transl Med Date: 2018-11-07 Impact factor: 17.956