Metabolomics highlights cardiovascular risk factor links to sphingolipid and one-carbon metabolism in asymptomatic adults

Background: Metabolic perturbations associated with cardiovascular disease (CVD) risk factors prior to clinical disease manifestation may inform novel preventative measures. Here we characterize metabolome-wide associations with various cardiometabolic risk factors in an asymptomatic population to develop a mechanistic metabolic framework of preemergent metabolic shifts. Methods: Plasma was collected from a clinically healthy cross-sectional cohort (n = 361; Clinical Trials.gov ID: NCT02367287). Cohort mass spectrometry-based metabolomics (biocrates MxP(R) Quant 500 kits) were complemented with expanded lipidomic coverage for a sex/age-matched subset with LDLc difference > 60 mg/dL (n = 38). LDLc-focused partial least square regressions and discriminate analyses were performed, after false discovery rate corrections. Results: As LDLc levels increased, plasma sphingolipids and cholesteryl esters, and transulfuration products were higher, while ether-linked phosphatidylcholines serine, glycine, alanine, methionine were lower. The subset analysis further highlighted elevations in triglycerides and deoxyceramides with elevated LDLc. When adjusted for alanine concentrations, deoxyceramide concentrations increased as serine declined (p = 0.0012), and the deoxyceramide:ceramide ratios were higher in the high LDLc group (p = 0.039). Lower serine levels were also accompanied by parallel changes in glycine (p= 0.0004), and anti-parallel changes in transsulfuration pathway metabolites particularly when adjusted for ceramide levels. Notably, the alanine:serine ratio was more strongly associated with fasting triglycerides (r = 0.38; p <0.0001) than LDLc. Conclusions: Serine depletion by increased demands of sphingolipid, and possibly transsulfuration pathway, metabolism is associated with elevations in cytotoxic deoxyceramides and reductions in glycine and methionine availability for one-carbon metabolism. These changes were strongly associated with fasting triglycerides and insulin resistance, a key marker of residual CVD risk.