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Abstract
In the last decade, clinical studies have investigated the clinical relevance of circulating cell-free-DNA (ccfDNA) as a diagnostic and prognosis tool in various diseases including cancers. However, limited knowledge on ccfDNA biology restrains its full development in the clinical practice. To improve our understanding, we evaluated the impact of the circadian rhythm on ccfDNA release in healthy subjects over a 24-h period. 10 healthy female subjects underwent blood sampling at 8am and 20 healthy male subjects underwent serial blood sampling (8:00 AM, 9:00 AM, 12:00 PM, 4:00 PM, 8:00 PM, 12:00 AM, 4 AM (+ 1 Day) and 8 AM (+ 1 Day)). We performed digital droplet-based PCR (ddPCR) assays to target 2 DNA fragments (69 & 243 bp) located in the KRAS gene to determine the ccfDNA concentration and fragmentation profile. As control, half of the samples were re-analyzed by capillary miniaturized electrophoresis (BIAbooster system). Overall, we did not detect any influence of the circadian rhythm on ccfDNA release. Instead, we observed a decrease in the ccfDNA concentration after meal ingestion, suggesting either a post-prandial effect or a technical detection bias due to a higher plasma load in lipids and triglycerides. We also noticed a potential effect of gender, weight and creatinine levels on ccfDNA concentration.
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1 Université de Paris, UMR-S1138, CNRS SNC5096, Équipe Labélisée Ligue Nationale Contre le Cancer, Centre de Recherche des Cordeliers, Paris, France (GRID:grid.410511.0) (ISNI:0000 0001 2149 7878); Eurofins-Biomnis, Gerland, Lyon, France (GRID:grid.410511.0)
2 CIC1418 and DMU CARTE, AP-HP, Hôpital Européen Georges-Pompidou, Paris, France (GRID:grid.414093.b) (ISNI:0000 0001 2183 5849)
3 Université de Paris, UMR-S1138, CNRS SNC5096, Équipe Labélisée Ligue Nationale Contre le Cancer, Centre de Recherche des Cordeliers, Paris, France (GRID:grid.410511.0) (ISNI:0000 0001 2149 7878)
4 Adelis, Labège, France (GRID:grid.410511.0)
5 Eurofins-Biomnis, Gerland, Lyon, France (GRID:grid.410511.0)
6 Université de Paris, UMR-S1138, CNRS SNC5096, Équipe Labélisée Ligue Nationale Contre le Cancer, Centre de Recherche des Cordeliers, Paris, France (GRID:grid.410511.0) (ISNI:0000 0001 2149 7878); Hôpital Européen Georges Pompidou (HEGP), Assistance Publique Hôpitaux de Paris (AP-HP), Biochemistry Department - Unit of Pharmacogenetic and Molecular Oncology, Paris, France (GRID:grid.414093.b) (ISNI:0000 0001 2183 5849)
7 Université de Paris, UMR-S1138, CNRS SNC5096, Équipe Labélisée Ligue Nationale Contre le Cancer, Centre de Recherche des Cordeliers, Paris, France (GRID:grid.410511.0) (ISNI:0000 0001 2149 7878); Gustave Roussy, Department of Visceral and Surgical Oncology, Villejuif, France (GRID:grid.14925.3b) (ISNI:0000 0001 2284 9388)