Integrative Multiparametric Analysis of Circulating Cell‐Free Nucleic Acids of Plasma in Healthy Individuals During Aging

• Published in: Aging cell Vol. 24; no. 9; pp. e70133 - n/a
• Main Authors: Tessier, Nicolas P., Hardy, Lise M., Mauger, Florence, Daunay, Antoine, Daviaud, Christian, Hchaichi, Ilef, Horgues, Caroline, Sahbatou, Mourad, Le Buanec, Hélène, Deleuze, Jean‐François, How‐Kit, Alexandre
• Format: Journal Article
• Language: English
• Published: England John Wiley & Sons, Inc 01.09.2025; Wiley Open Access
• Subjects: Adult; Age groups; Aged; Aging; Aging - blood; Aging - genetics; Analysis; Biochemistry, Molecular Biology; Biomarkers; Blood banks; Capillary electrophoresis; Cell-Free Nucleic Acids - blood; circulating cell‐free DNA; circulating cell‐free nucleic acids; circulating cell‐free RNA; DNA Methylation; Ethical aspects; Ethylenediaminetetraacetic acid; Female; Genomics; Humans; Leukocytes; Life Sciences; Male; Methylation; Middle Aged; miRNA; Mitochondrial DNA; Nucleotide sequence; Older people; Plasma; RNA; Tropical diseases; Young Adult; France; Paris France; circulating cell‐free DNA; circulating cell‐free nucleic acids; DNA methylation; circulating cell‐free RNA; miRNA; aging; plasma; aging circulating cell-free DNA circulating cell-free nucleic acids circulating cell-free RNA DNA methylation miRNA plasma; circulating cell-free RNA; circulating cell-free nucleic acids; circulating cell-free DNA
• ISSN: 1474-9718; 1474-9726; 1474-9726; 1474-9728
• DOI: 10.1111/acel.70133

ABSTRACT Plasma circulating cell‐free nucleic acids (ccfNAs) provide an exceptional source of information about an individual's health, yet their biology in healthy individuals during aging remains poorly understood. Here, we present the first integrative multiparametric analysis of the major types of plasma ccfNAs, including nuclear (ccfnDNA) and mitochondrial (ccfmtDNA) DNA, as well as ribosomal (ccfrRNA), messenger (ccfmRNA) and micro‐RNA (ccfmiRNA) in 139 healthy donors aged 19–66 years. We focused on quantity, integrity, and DNA methylation using an optimized experimental workflow that combines highly sensitive analytical methods with the detection of highly repetitive DNA and highly abundant RNA sequences, thereby reducing the required amount of ccfNAs per analysis. We showed a highly significant increase in ccfnDNA levels during aging (p < 0.001), associated with a decrease in its integrity (p < 0.05), while no significant changes were detected in ccfmtDNA levels and ccfDNA methylation. Moreover, a significant increase in ccfmRNA and ccfrRNA (p < 0.05), as well as miR‐483‐5p (p < 0.001) levels was detected during aging, but without any changes in ccfRNA integrity. Finally, we also showed that ccfDNA and ccfRNA levels were correlated (p < 0.001), and a similar pattern was observed for ccfmtDNA and ccfRNA levels, suggesting a possible common release, maintenance, and/or clearance mechanism. Therefore, our study provides an optimized workflow for the global analysis of ccfNAs, enhances the understanding of their biology during aging, and identifies several potential ccfNA‐based biomarkers of aging. We performed the first integrative multiparametric analysis of plasma circulating cell‐free nucleic acids, including nuclear and mitochondrial DNA, as well as ribosomal, messenger, and micro‐RNAs, in a cohort of 139 healthy individuals. We focused on quantity, integrity, and DNA methylation using assays targeting highly repetitive DNA and highly abundant RNA.