Exploration of Human Serum Lipoprotein Supramolecular Phospholipids Using Statistical Heterospectroscopy in n-Dimensions (SHY-n): Identification of Potential Cardiovascular Risk Biomarkers Related to SARS-CoV-2 Infection

Abstract

SARS-CoV-2 infection causes a significant reduction in lipoprotein-bound serum phospholipids give rise to supramolecular phospholipid composite (SPC) signals observed in diffusion and relaxation edited 1H NMR spectra. To characterize the chemical structural components and compartmental location of SPC and to understand further its possible diagnostic properties, we applied a Statistical HeterospectroscopY in n-dimensions (SHY-n) approach. This involved statistically linking a series of orthogonal measurements made on the same samples, using independent analytical techniques and instruments, to identify the major individual phospholipid components giving rise to the SPC signals. Thus, an integrated model for SARS-CoV-2 positive and control adults is presented that relates three identified diagnostic subregions of the SPC signal envelope (SPC1, SPC2, and SPC3) generated using diffusion and relaxation edited (DIRE) NMR spectroscopy to lipoprotein and lipid measurements obtained by in vitro diagnostic NMR spectroscopy and ultrahigh-performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS). The SPC signals were then correlated sequentially with (a) total phospholipids in lipoprotein subfractions; (b) apolipoproteins B100, A1, and A2 in different lipoproteins and subcompartments; and (c) MS-measured total serum phosphatidylcholines present in the NMR detection range (i.e., PCs: 16.0,18.2; 18.0,18.1; 18.2,18.2; 16.0,18.1; 16.0,20.4; 18.0,18.2; 18.1,18.2), lysophosphatidylcholines (LPCs: 16.0 and 18.2), and sphingomyelin (SM 22.1). The SPC3/SPC2 ratio correlated strongly (r = 0.86) with the apolipoprotein B100/A1 ratio, a well-established marker of cardiovascular disease risk that is markedly elevated during acute SARS-CoV-2 infection. These data indicate the considerable potential of using a serum SPC measurement as a metric of cardiovascular risk based on a single NMR experiment. This is of specific interest in relation to understanding the potential for increased cardiovascular risk in COVID-19 patients and risk persistence in post-acute COVID-19 syndrome (PACS).

Direct low field J-edited diffusional proton NMR spectroscopic measurement of COVID-19 inflammatory biomarkers in human serum

“…S3B †), which stems from LDL lipoproteins is almost completely eliminated at 298 K compared to the low frequency part of SPC, stemming from HDL lipoproteins. 31 Similar to SPC, Glyc is also reduced by ∼20% in intensity at lower temperature (Fig. S3C †).…”

“…In addition, the different regions of SPC are affected differently. The high frequency part of SPC, previously referred to as SPC-B 31 (Figure S3B), which stems from LDL lipoproteins is almost completely eliminated at 298 K compared to the low frequency part of SPC, SPC-A stemming from HDL lipoproteins 31 . Similar to SPC, Glyc is also reduced by ~20 % in intensity at lower temperature (Figure S3C).…”

“…SARS-CoV-2-(+) patients could be effectively distinguished from healthy controls modelling untargeted 1 H NMR data (Figure 1A). Phenoconversion from a healthy into an infected stated is associated with changes in the concentrations of a wide range of metabolic entities (lipoproteins, glycoproteins, amino acids, lipids and other metabolites) that can be derived from NMR spectroscopic and mass spectrometric data [26][27][28] , and which result in distinctive embedded biomarker features including some of those previously observed in diabetes 29,30 , cardiovascular disease [31][32][33][34][35] , liver dysfunction 36,37 , neurological disruption 38 and inflammation 39,40 .Many of these pathological changes can be observed using a variety of NMR experiments on blood plasma and these changes are robust to sample handling with standardised protocols 41,42 and were also observed by others 22,[43][44][45] .…”

Lysophosphatidylcholine: Potential Target for the Treatment of Chronic Pain

“…At present, the accurate detection of LPC relies on metabolomics or lipidomics technology, which provides assays for exploiting the role of LPC in chronic pain. Apart from chronic pain, LPC or LPC species in body fluids such as blood, urine, cerebrospinal fluid, and tissues are uniquely or collectively related to cancer [ 150 , 151 , 152 ], diabetes [ 153 , 154 , 155 , 156 ], coronary atherosclerosis [ 157 ], Alzheimer’s disease [ 158 , 159 ], rheumatoid arthritis [ 83 ], COVID-19 [ 160 ], liver and kidney damage [ 161 , 162 ], etc. Whether LPC is necessary for other chronic pain conditions, such as cancer pain, has not been confirmed.…”

Metabolomics as a powerful tool for diagnostic, pronostic and drug intervention analysis in COVID-19

“…Unfortunately, COVID-19 does not always evolve towards a full restoration of the metabolism and post-acute sequelae of COVID-19 represent an emerging global crisis ( Su et al, 2022 ) and the need to find biomarkers for long COVID is pressing. In this context, deconvolution of NMR spectra from COVID-19 sera identified three diagnostic subregions of the supramolecular phospholipid composite signal envelope that provide insight about the increased cardiovascular risk in COVID-19 patients and the risk persistence in post-acute COVID-19 syndrome ( Masuda et al, 2022 ). Notably, lipoproteins emerge as an important prognostic biomarker for the prediction of long COVID effects ( Bai et al, 2021 ).…”