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Characterization of the cellular lipid composition during SARS-CoV-2 infection

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Abstract

Emerging and re-emerging zoonotic viral diseases continue to significantly impact public health. Of particular interest are enveloped viruses (e.g., SARS-CoV-2, the causative pathogen of COVID-19), which include emerging pathogens of highest concern. Enveloped viruses contain a viral envelope that encapsulates the genetic material and nucleocapsid, providing structural protection and functional bioactivity. The viral envelope is composed of a coordinated network of glycoproteins and lipids. The lipid composition of the envelope consists of lipids preferentially appropriated from host cell membranes. Subsequently, changes to the host cell lipid metabolism and an accounting of what lipids are changed during viral infection provide an opportunity to fingerprint the host cell’s response to the infecting virus. To address this issue, we comprehensively characterized the lipid composition of VeroE6-TMPRSS2 cells infected with SARS-CoV-2. Our approach involved using an innovative solid-phase extraction technique to efficiently extract cellular lipids combined with liquid chromatography coupled to high-resolution tandem mass spectrometry. We identified lipid changes in cells exposed to SARS-CoV-2, of which the ceramide to sphingomyelin ratio was most prominent. The identification of a lipid profile (i.e., lipid fingerprint) that is characteristic of cellular SARS-CoV-2 infection lays the foundation for targeting lipid metabolism pathways to further understand how enveloped viruses infect cells, identifying opportunities to aid antiviral and vaccine development.

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Abbreviations

WHO:

World Health Organization

HIV:

Human immunodeficiency virus

HSV:

Herpes simplex virus

RSV:

Respiratory syncytial virus

VSV:

Vesicular stomatitis virus

LC HR-MS/MS:

Liquid chromatography, high-resolution tandem mass spectrometry

SPE:

Solid-phase extraction

HPLC:

High-pressure liquid chromatography

MTBE:

Tert-Butyl methyl ether

HRMS:

High-resolution mass spectrometry

Q-TOF:

Quadrupole time of flight

LLE:

Liquid-liquid extraction

PCA:

Principal component analysis

PLS-DSA:

Partial least squares discriminate analysis

CE:

Cholesterol esters

GP:

Glycerophospholipids

DG:

Diacylglycerol

TG:

Triacylglycerol

SP:

Sphingolipids

Cer:

Ceramide

SM:

Sphingomyelin

LPC:

Lysophosphocholine

LPE:

Lysophosphoethanolamine

PC:

Glycerophosphoscholine

ePC:

Ether glycerophosphocholine

PE :

Glycerophosphoethanolamine

PI:

Glycerophosphoinositol

PS:

Glycerophosphoserine

IVA:

Influenza A

VE6-TMPRSS2:

Vero E6 cells overexpressing TMPRSS2

ASM:

Acid sphingomyelinase

FDA:

Federal Drug Administration

FIASMAs:

Functional inhibitors of acid sphingomyelinase

PLA2 :

Phospholipase A2

cPLA2 :

Cytosolic PLA2

SEM:

Standard error of the mean

TIC:

Total ion chromatogram

EIC:

Extracted ion chromatogram

t R :

Retention time

FC:

Fold change

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Acknowledgements

The authors acknowledge the University of Maryland School of Pharmacy Faculty Start-up funds (J.W.J.); Agilent Research Gift #4520 (J.W.J.); University of Maryland, Baltimore, Institute for Clinical & Translational Research (ICTR) and the National Center for Advancing Translational Sciences (NCATS) Clinical Translational Science Award (CTSA) grant number 1UL1TR003098 (J.W.J.); University of Maryland School of Pharmacy Mass Spectrometry Center (SOP1841-IQB2014) and Cornell University NIH/NIAID grants R01 IA109022 and R21 AI142377 (H.A.C.); Emergent Ventures at Mercatus Center, George Mason Univ. Fast Grant # 2103 (H.A.C.); and Cornell Seed grants (H.A.C.).

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, 20 N. Pine Street, Room N721, Baltimore, MD, 21201, USA

    Ahmed M. Abdel-Megied & Jace W. Jones

  2. Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Kafr El-Sheikh University, Kafr El-Sheikh City, Egypt

    Ahmed M. Abdel-Megied

  3. Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY, 14853, USA

    Isaac A. Monreal & Hector C. Aguilar

  4. Agilent Technologies, Wilmington, DE, USA

    Limian Zhao

  5. Agilent Technologies, Santa Clara, CA, USA

    Alex Apffel

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Abdel-Megied, A.M., Monreal, I.A., Zhao, L. et al. Characterization of the cellular lipid composition during SARS-CoV-2 infection. Anal Bioanal Chem 415, 5269–5279 (2023). https://doi.org/10.1007/s00216-023-04825-1

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