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Updated: Jul 22, 2025
Cell-free Biochemical Fluorometric Enzymatic Assay for High-throughput Measurement of Lipid Peroxidation in High Density Lipoprotein
Published on: October 12, 2017
Cell-free, high-density lipoprotein-specific phospholipid efflux assay predicts incident cardiovascular disease
Masaki Sato1,2,3, Edward B Neufeld1, Martin P Playford4
1Lipoprotein Metabolism Laboratory, National Heart, Lung, and Blood Institute (NHLBI), NIH, Bethesda, Maryland, USA.
Insights
A new HDL-specific phospholipid efflux (HDL-SPE) assay shows strong associations with coronary artery disease (CAD) and predicts cardiovascular disease (CVD) events. This functional HDL assay offers improved risk assessment beyond traditional markers.
Area of Science:
- Cardiovascular Disease Research
- Biomarker Development
- Lipid Metabolism
Background:
- Cellular cholesterol efflux capacity (CEC) predicts cardiovascular disease (CVD) events but is not clinically feasible.
- High-density lipoprotein (HDL) cholesterol (HDL-C) levels are insufficient for accurate CVD risk stratification.
- Novel assays are needed to assess HDL functionality for improved CVD risk assessment.
Purpose of the Study:
- To develop and validate a novel HDL-specific phospholipid efflux (HDL-SPE) assay.
- To assess the association of HDL-SPE with prevalent coronary artery disease (CAD).
- To evaluate the predictive value of HDL-SPE for incident CVD events.
Main Methods:
- Developed an HDL-SPE assay measuring HDL apolipoprotein-mediated phospholipid solubilization.
- Assessed HDL-SPE in two independent cohorts with prevalent CAD (n=120 and n=224).
- Examined HDL-SPE association with incident CVD events in the PREVEND study (n=680) matched for risk factors.
Main Results:
- HDL-SPE demonstrated stronger associations with prevalent CAD than HDL-C or apolipoprotein A-I (apoA-I) in both studies.
- Area under the curve (AUC) for HDL-SPE in predicting CAD was significantly higher (Study I: 0.68, Study II: 0.83).
- HDL-SPE independently predicted incident CVD events in the PREVEND study (OR < 0.2 per SD increment, P < 0.001).
Conclusions:
- The HDL-SPE assay is a robust measure of HDL functionality.
- HDL-SPE shows significant potential for improving cardiovascular risk assessment.
- This assay could facilitate drug discovery targeting HDL function.
Abstract:
BACKGROUNDCellular cholesterol efflux capacity (CEC) is a better predictor of cardiovascular disease (CVD) events than HDL-cholesterol (HDL-C) but is not suitable as a routine clinical assay.METHODSWe developed an HDL-specific phospholipid efflux (HDL-SPE) assay to assess HDL functionality based on whole plasma HDL apolipoprotein-mediated solubilization of fluorescent phosphatidylethanolamine from artificial lipid donor particles. We first assessed the association of HDL-SPE with prevalent coronary artery disease (CAD): study I included NIH severe-CAD (n = 50) and non-CAD (n = 50) participants, who were frequency matched for sex, BMI, type 2 diabetes mellitus, and smoking; study II included Japanese CAD (n = 70) and non-CAD (n = 154) participants. We also examined the association of HDL-SPE with incident CVD events in the Prevention of Renal and Vascular End-stage Disease (PREVEND) study comparing 340 patients with 340 controls individually matched for age, sex, smoking, and HDL-C levels.RESULTSReceiver operating characteristic curves revealed stronger associations of HDL-SPE with prevalent CAD. The AUCs in study I were as follows: HDL-SPE, 0.68; apolipoprotein A-I (apoA-I), 0.62; HDL-C, 0.63; and CEC, 0.52. The AUCs in study II were as follows: HDL-SPE, 0.83; apoA-I, 0.64; and HDL-C, 0.53. Also longitudinally, HDL-SPE was significantly associated with incident CVD events independent of traditional risk factors with ORs below 0.2 per SD increment in the PREVEND study (P < 0.001).CONCLUSIONHDL-SPE could serve as a routine clinical assay for improving CVD risk assessment and drug discovery.TRIAL REGISTRATIONClinicalTrials.gov NCT01621594.FUNDINGNHLBI Intramural Research Program, NIH (HL006095-06).
