Age-Associated Metabolomic Changes in Human Spermatozoa

The functional genomic mechanisms contributing to aging-associated decline in fertility in men remain insufficiently elucidated. This study investigated age-related alterations in the sperm metabolome of healthy fertile Arab men across three groups: young adult (21–30 years, n = 6), late adult (31–40 years, n = 7), and advanced age (41–51 years, n = 5). Metabolomics was performed using LC-MS/MS. Statistical/functional analyses were performed using MetaboAnalyst-Pro. A total of 380 metabolites were identified, of which 164 showed significant differences (p < 0.05) across age groups. Principal component analysis, partial least squares-discriminate (PLS-DA), and sparse PLS-DA consistently demonstrated distinct metabolomic clustering between young adult and advanced age groups. Notably, in the advanced-age spermatozoa, L-homocysteine was undetectable, while methyloctadecanoyl-CoA was uniquely abundant. Biomarker analysis identified 137 potential aging-sperm biomarkers (AUC = 1), including upregulated (e.g., pentadecanoyl-CoA, (3S)-3-hydroxylinoleoyl-CoA, CDP-DG(LTE4/20:4(8Z11Z14Z17Z)), uracil) and downregulated (e.g., (S)-hydroxyoctanoyl-CoA, DG(22:6/18:4), L-homocysteine, N-myristoyl serine) metabolites. These biomarkers participate in key sperm domains, including motility, energy metabolism, membrane remodeling, oxidative-stress regulation, and fertilization. In conclusion, advancing age disrupts sperm “metabolostasis” (metabolite homeostasis essential for normal function), compromising their physiological integrity and fertilization competence. The identified biomarkers offer promising targets for interventions to preserve sperm health and mitigate age-related fertility decline.