Characterization of Plasma Cell-Free DNA Integrity Using Droplet-Based Digital PCR: Toward the Development of Circulating Tumor DNA-Dedicated Assays

Autor: Geoffroy Poulet, Fanny Garlan, Sonia Garrigou, Eleonora Zonta, Leonor Benhaim, Marie-Jennifer Carrillon, Audrey Didelot, Delphine Le Corre, Claire Mulot, Philippe Nizard, Frederic Ginot, Audrey Boutonnet-Rodat, Helene Blons, Jean-Baptiste Bachet, Julien Taïeb, Aziz Zaanan, Vanna Geromel, Laurence Pellegrina, Pierre Laurent-Puig, Shu-Fang Wang-Renault, Valerie Taly
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Zdroj: Frontiers in Oncology, Vol 11 (2021)
Druh dokumentu: article
ISSN: 2234-943X
DOI: 10.3389/fonc.2021.639675
Popis: Background: Cellular-cell free-DNA (ccfDNA) is being explored as a diagnostic and prognostic tool for various diseases including cancer. Beyond the evaluation of the ccfDNA mutational status, its fragmentation has been investigated as a potential cancer biomarker in several studies. However, probably due to a lack of standardized procedures dedicated to preanalytical and analytical processing of plasma samples, contradictory results have been published.Methods: ddPCR assays allowing the detection of KRAS wild-type and mutated sequences (KRAS p.G12V, pG12D, and pG13D) were designed to target different fragments sizes. Once validated on fragmented and non-fragmented DNA extracted from cancer cell lines, these assays were used to investigate the influence of the extraction methods on the non-mutated and mutated ccfDNA integrity reflected by the DNA integrity index (DII). The DII was then analyzed in two prospective cohorts of metastatic colorectal cancer patients (RASANC study n = 34; PLACOL study n = 12) and healthy subjects (n = 49).Results and Discussion: Our results demonstrate that ccfDNA is highly fragmented in mCRC patients compared with healthy individuals. These results strongly suggest that the characterization of ccfDNA integrity hold great promise toward the development of a universal biomarker for the follow-up of mCRC patients. Furthermore, they support the importance of standardization of sample handling and processing in such analysis.
Databáze: Directory of Open Access Journals