Cell-free, high-density lipoprotein–specific phospholipid efflux assay predicts incident cardiovascular disease
Masaki Sato, … , Uwe J.F. Tietge, Alan T. Remaley
Masaki Sato, … , Uwe J.F. Tietge, Alan T. Remaley
Published July 20, 2023
Citation Information: J Clin Invest. 2023;133(18):e165370. https://doi.org/10.1172/JCI165370.
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Clinical Medicine Vascular biology

Cell-free, high-density lipoprotein–specific phospholipid efflux assay predicts incident cardiovascular disease

Abstract

BACKGROUND Cellular 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.METHODS We 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.RESULTS Receiver 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).CONCLUSION HDL-SPE could serve as a routine clinical assay for improving CVD risk assessment and drug discovery.TRIAL REGISTRATION ClinicalTrials.gov NCT01621594.FUNDING NHLBI Intramural Research Program, NIH (HL006095-06).

Authors

Masaki Sato, Edward B. Neufeld, Martin P. Playford, Yu Lei, Alexander V. Sorokin, Angel M. Aponte, Lita A. Freeman, Scott M. Gordon, Amit K. Dey, Kianoush Jeiran, Masato Hamasaki, Maureen L. Sampson, Robert D. Shamburek, Jingrong Tang, Marcus Y. Chen, Kazuhiko Kotani, Josephine L.C. Anderson, Robin P.F. Dullaart, Nehal N. Mehta, Uwe J.F. Tietge, Alan T. Remaley

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Figure 1

Efflux of fluorescent PE from donor LC-CSH particles to plasma lipoproteins is HDL specific.

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Efflux of fluorescent PE from donor LC-CSH particles to plasma lipoprote...
(A) Model of cell-free HDL-SPE and NS-CEC assays, as described in Methods. After centrifugation, lipoprotein-associated fluorescence in the supernatant is measured by agarose gel electrophoresis and fluorometry. HDL acquires both *PE and *Chol, whereas LDL/VLDL acquires only *Chol. (B) Effect of pooled normal HP volume on fluorescent lipid efflux. Agarose gel electrophoresis: *PE transfer from donor particles was HDL specific, whereas *Chol transfer to plasma lipoproteins was nonspecific. (C) Fluorometry of the reaction mixture supernatant revealed that the dependence of *PE and *Chol efflux on plasma volume differed. (D) Both HDL-SPE and NS-CEC were linear using 15–35 μL plasma. (E) Fluorometric and electrophoretic gel analyses of HDL-SPE were highly correlated. (F) LC-CSH *PE robustly effluxed to isolated HDL, whereas little to no *PE effluxed to isolated LDL or VLDL. All data are presented as the mean ± SD from triplicate assays unless otherwise stated. A P value of less than 0.05 was considered statistically significant. SUP, supernatant; R, released.