Lipid-Based Nutrient Supplementation Increases High-Density Lipoprotein (HDL) Cholesterol Efflux Capacity and Is Associated with Changes in the HDL Glycoproteome in Children

Abstract

Prenatal plus postnatal small-quantity lipid-based nutrient supplements (SQ-LNS) improved child growth at 18 months in the International Lipid-Based Nutrient Supplements DYAD trial in Ghana. In this secondary outcome analysis, we determined whether SQ-LNS versus prenatal iron and folic acid (IFA) supplementation improves the cholesterol efflux capacity (CEC) of high-density lipoprotein (HDL) particles and alters their lipidomic, proteomic, or glycoproteomic composition in a subset of 80 children at 18 months of age. HDL CEC was higher among children in the SQ-LNS versus IFA group (20.9 ± 4.1 vs 19.4 ± 3.3%; one-tailed p = 0.038). There were no differences in HDL lipidomic or proteomic composition between groups. Twelve glycopeptides out of the 163 analyzed were significantly altered by SQ-LNS, but none of the group differences remained significant after correction for multiple testing. Exploratory analysis showed that 6 out of the 33 HDL-associated proteins monitored differed in glycopeptide enrichment between intervention groups, and 6 out of the 163 glycopeptides were correlated with CEC. We conclude that prenatal plus postnatal SQ-LNS may modify HDL protein glycoprofiles and improve the CEC of HDL particles in children, which may have implications for subsequent child health outcomes. This trial was registered at as NCT00970866.

High-Density Lipoprotein Changes in Alzheimer’s Disease Are APOE Genotype-Specific

“…HDL cholesterol efflux was measured using a commercially available kit (Abcam, Cambridge, UK, catalog ab19685) with modifications, as previously described [ 14 ]. J774A.1 (ATCC, Manassas, VA, USA, catalog TIB-67) were seeded at 100,000 cells in 96-well plates for 4 h in Dulbecco’s Modified Eagle’s Medium containing 10% fetal bovine serum and 100 μg/mL penicillin and streptomycin.…”

Glycosylation of HDL-Associated Proteins and Its Implications in Cardiovascular Disease Diagnosis, Metabolism and Function

“…Because HDL particles are so small (7-12 nm in diameter) as to overlap with many plasma proteins in terms of their size (e.g., ferritin), and because they are close in density to other lipoprotein particles and even extracellular vesicles, they are difficult to isolate and purify. According to multiple proteomic studies HDL could carry as few as 12 key proteins or up to an excess of 200 proteins (24,25) depending on how they are isolated (24,26,27). Various methods, and combinations of these methods, have been used to isolate HDL including ultracentrifugation, size exclusion chromatography, immunoaffinity precipitation, and asymmetrical flow field flow fractionation.…”

“…ApoA-I, the major structural, defining HDL apoprotein accounting for around 70% of total HDL protein mass, plays a key role in lipid and cholesterol metabolism and is highly associated with cardioprotection (42). Despite ApoA-I being reported to possibly be glycosylated (43)(44)(45) extensive mass spectrometry-based (MS)-based profiling demonstrated that there is no evidence of ApoA-I glycosylation (25). ApoA-I does not have the consensus sequence for N-glycosylation (AsnXxxSer/Thr/Cys, where Xxx can be any amino acid except proline), and whereas O-glycosylation is possible on any serine or threonine residue, detailed analysis of isolated HDL has not yielded any confirmed O-glycosylated peptides on ApoA-I.…”

“…ApoA-II is the second most abundant HDL apoprotein, representing as much as 20% of total protein mass (42), and has been shown to have important implications for CV health though results were historically inconsistent and controversial. For instance one early study showed that low serum ApoA-II was a marker of atheroprotection in patients with non-insulindependent-diabetes mellitus (46) but conversely another study showed that elevated levels of ApoA-II were proatherogenic Proteins included in this table include only those with glycosylation sites confirmed to actually express glycans at those sites by mass spectrometry analysis of isolated HDL fractions from a starting volume of 500 uL of plasma, as described in (25). HDL-associated proteins that have been reported to be glycosylated previously, and/or have putative sites but that either could not be confirmed by mass spectrometry or are present at low abundance such that they fall under the limit of detection, are not reported in this table.…”

Quantitative glycoproteomics of high-density lipoproteins

“…Target discovery of HDL-associated proteins was accomplished through bottom-up glycoproteomics using Orbitrap MS on a total of 80 samples from young children. 15 Proteomics of HDL and other nanoparticles have been performed previously. 5,11,[16][17][18] We considered both N-and O-linked glycosylation in searching for potential glycopeptides.…”