NMR Spectroscopic Windows on the Systemic Effects of SARS-CoV-2 Infection on Plasma Lipoproteins and Metabolites in Relation to Circulating Cytokines

Abstract

To investigate the systemic metabolic effects of SARS-CoV-2 infection, we analyzed 1 H NMR spectroscopic data on human blood plasma and co-modeled with multiple plasma cytokines and chemokines (measured in parallel). Thus, 600 MHz 1 H solvent-suppressed single-pulse, spin-echo, and 2D J-resolved spectra were collected on plasma recorded from SARS-CoV-2 rRT-PCR-positive patients (n = 15, with multiple sampling timepoints) and agematched healthy controls (n = 34, confirmed rRT-PCR negative), together with patients with COVID-19/influenza-like clinical symptoms who tested SARS-CoV-2 negative (n = 35). We compared the single-pulse NMR spectral data with in vitro diagnostic research (IVDr) information on quantitative lipoprotein profiles (112 parameters) extracted from the raw 1D NMR data. All NMR methods gave highly significant discrimination of SARS-CoV-2 positive patients from controls and SARS-CoV-2 negative patients with individual NMR methods, giving different diagnostic information windows on disease-induced phenoconversion. Longitudinal trajectory analysis in selected patients indicated that metabolic recovery was incomplete in individuals without detectable virus in the recovery phase. We observed four plasma cytokine clusters that expressed complex differential statistical relationships with multiple lipoproteins and metabolites. These included the following: cluster 1, comprising MIP-1β, SDF-1α, IL-22, and IL-1α, which correlated with multiple increased LDL and VLDL subfractions; cluster 2, including IL-10 and IL-17A, which was only weakly linked to the lipoprotein profile; cluster 3, which included IL-8 and MCP-1 and were inversely correlated with multiple lipoproteins. IL-18, IL-6, and IFN-γ together with IP-10 and RANTES exhibited strong positive correlations with LDL1−4 subfractions and negative correlations with multiple HDL subfractions. Collectively, these data show a distinct pattern indicative of a multilevel cellular immune response to SARS CoV-2 infection interacting with the plasma lipoproteome giving a strong and characteristic immunometabolic phenotype of the disease. We observed that some patients in the respiratory recovery phase and testing virus-free were still metabolically highly abnormal, which indicates a new role for these technologies in assessing full systemic recovery.

Molecular Phenomic Approaches to Deconvolving the Systemic Effects of SARS-CoV-2 Infection and Post-acute COVID-19 Syndrome

“…It is essentially a systemic metabolic disease driven by the heightened activities of the immune system responses to the viral presence. Profound effects are seen in lipid (Kimhofer et al 2020;Chen et al 2020) and amino acid (Wu et al 2020b;Cai et al 2020;Lawler et al 2021) metabolism and other small molecules as well as shifts in lipoprotein profiles, these changes reflecting liver, cardiovascular, diabetic, muscle wasting, and neuropathological effects (Kimhofer et al 2020;Chen et al 2020;Wu et al 2020b;Cai et al 2020;Lawler et al 2021;Lodge et al 2021a). There is also a plethora of other effects including anosmia, Guillain-Barre syndrome, and chronic fatigue) and most of those symptoms can be persistent in patients with PACS (Nalbandian et al 2021).…”

“…The various spectroscopic "windows" into the disease process reveal different, but complementary, insights into the altered biochemistry and pathological processes. NMR spectroscopy of blood plasma has given new metabolic, glycoproteomic and lipoproteomic insights into the acute COVID-19 disease process and the relationships between these parameters and cytokines and chemokines indicates the presence of an activated immune-metabolic axis (Kimhofer et al 2020;Lawler et al 2021;Lodge et al 2021a). Even simple metabolic measurements can be revealing, thus, in severely affected patients, the plasma lactate to pyruvate ratio can be high reflecting impaired peripheral blood oxygenation and poor pulmonary function (Kimhofer et al 2020).…”

“…Proton NMR methods allow simultaneous observation of metabolites, lipoproteins and glycoproteins (Nicholson et al 2002(Nicholson et al , 2012. Thus, acute-phase reactive proteins such as a-1acid glycoprotein (GlycA) are highly elevated in plasma in active COVID-19 disease, and there is also a marked reduction in HDL-related species, e.g., phospholipids, with a significant rise in many VLDL and LDL components (Kimhofer et al 2020;Lodge et al 2021a). In particular, the Apolipoprotein B100/Apolipoprotein A1 ratio (ABA1) is increased in active COVID-19 disease and ABA1 values have previously been associated with increased cardiovascular and atherosclerotic risks (Holmes et al 2008b;Lodge et al 2021a).…”

Incomplete Systemic Recovery and Metabolic Phenoreversion in Post-Acute-Phase Nonhospitalized COVID-19 Patients: Implications for Assessment of Post-Acute COVID-19 Syndrome

“… 23 − 28 We postulate that full functional recovery from the disease, by most of the patients without pre-existing comorbidities, is likely to be reflected in a normalization of the disrupted sentinel biomarker parameters and the overall metabolic phenotype. The process of multilevel recovery can be considered to be functionally equivalent to the reverse of disease onset and progression “phenoconversion”, 23 , 24 during which the measured metabolic profile shifts from normality to one typical of the disease or its subphenotypes. From a theoretical point of view, patient “phenoreversion” therefore would be partial or complete, reflecting the degree of systemic recovery from COVID-19 infection.…”

Diagnostic Potential of the Plasma Lipidome in Infectious Disease: Application to Acute SARS-CoV-2 Infection

“…Recent reports have detailed changes in plasma lipoproteins and the lipidome in response to SARS-CoV-2 infection [ 11 , 12 ]. Lipids are critical to systems biology, with essential roles as membrane structural components, signalling molecules, and energy sources, meaning that the lipidome is a rich source of biological information that has great potential in diagnostic pathology.…”