Auto-immunoproteomics analysis of COVID-19 ICU patients revealed increased levels of autoantibodies related to the male reproductive system

Frank Schmidt; Houari B Abdesselem; Karsten Suhre; Nishant N Vaikath; Muhammad U Sohail; Maryam Al-Nesf; Ilham Bensmail; Fathima Mashod; Hina Sarwath; Joerg Bernhardt; Stephanie Schaefer-Ramadan; Ti-Myen Tan; Priscilla E Morris; Edward J Schenck; David Price; Vidya Mohamed-Ali; Mohammed Al-Maadheed; Abdelilah Arredouani; Julie Decock; Jonathan M Blackburn; Augustine M K Choi; Omar M El-Agnaf

doi:10.3389/fphys.2023.1203723

Auto-immunoproteomics analysis of COVID-19 ICU patients revealed increased levels of autoantibodies related to the male reproductive system

Front Physiol. 2023 Jul 14:14:1203723. doi: 10.3389/fphys.2023.1203723. eCollection 2023.

Authors

Frank Schmidt¹, Houari B Abdesselem^{2

3}, Karsten Suhre⁴, Nishant N Vaikath³, Muhammad U Sohail¹, Maryam Al-Nesf^{5

6}, Ilham Bensmail², Fathima Mashod¹, Hina Sarwath¹, Joerg Bernhardt⁷, Stephanie Schaefer-Ramadan⁸, Ti-Myen Tan^{9

10

11}, Priscilla E Morris^{9

10

11}, Edward J Schenck¹², David Price¹², Vidya Mohamed-Ali^{6

13}, Mohammed Al-Maadheed^{6

13}, Abdelilah Arredouani^{14

15}, Julie Decock^{15

16}, Jonathan M Blackburn^{9

10

11}, Augustine M K Choi¹², Omar M El-Agnaf³

Affiliations

¹ Proteomics Core, Weill Cornell Medicine-Qatar, Doha, Qatar.
² Proteomics Core Facility, Qatar Biomedical Research Institute (QBRI), Qatar Foundation, Hamad Bin Khalifa University (HBKU), Doha, Qatar.
³ Neurological Disorders Research Center, QBRI, HBKU, Qatar Foundation, Doha, Qatar.
⁴ Bioinformatics Core, Weill Cornell Medicine-Qatar, Doha, Qatar.
⁵ Hamad General Hospital, Hamad Medical Corporation, Doha, Qatar.
⁶ Center of Metabolism and Inflammation, Division of Medicine, University College London, London, United Kingdom.
⁷ Institute for Microbiology, University of Greifswald, Greifswald, Germany.
⁸ Department of Genomic Medicine, Weill Cornell Medicine-Qatar, Doha, Qatar.
⁹ Department of Integrative Biomedical Sciences, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa.
¹⁰ Sengenics Corporation, Damansara Heights, Kuala Lumpur, Malaysia.
¹¹ Institute of Infectious Disease and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa.
¹² Department of Medicine, Division of Pulmonary and Critical Care Medicine, New York Presbyterian Hospital-Weill Cornell Medical Center, Weill Cornell Medicine, New York, NY, United States.
¹³ Anti-Doping Laboratory Qatar, Doha, Qatar.
¹⁴ Diabetes Research Center, Qatar Biomedical Research Institute (QBRI), Hamad Bin Khalifa University (HBKU), Qatar Foundation, Doha, Qatar.
¹⁵ College of Health and Life Sciences, Hamad Bin Khalifa University (HBKU), Qatar Foundation, Doha, Qatar.
¹⁶ Translational Cancer and Immunity Center, Qatar Biomedical Research Institute (QBRI), Hamad Bin Khalifa University (HBKU), Qatar Foundation, Doha, Qatar.

Abstract

Background: Coronavirus disease (COVID-19) manifests many clinical symptoms, including an exacerbated immune response and cytokine storm. Autoantibodies in COVID-19 may have severe prodromal effects that are poorly understood. The interaction between these autoantibodies and self-antigens can result in systemic inflammation and organ dysfunction. However, the role of autoantibodies in COVID-19 complications has yet to be fully understood. Methods: The current investigation screened two independent cohorts of 97 COVID-19 patients [discovery (Disc) cohort from Qatar (case = 49 vs. control = 48) and replication (Rep) cohort from New York (case = 48 vs. control = 28)] utilizing high-throughput KoRectly Expressed (KREX) Immunome protein-array technology. Total IgG autoantibody responses were evaluated against 1,318 correctly folded and full-length human proteins. Samples were randomly applied on the precoated microarray slides for 2 h. Cy3-labeled secondary antibodies were used to detect IgG autoantibody response. Slides were scanned at a fixed gain setting using the Agilent fluorescence microarray scanner, generating a 16-bit TIFF file. Group comparisons were performed using a linear model and Fisher's exact test. Differentially expressed proteins were used for KEGG and WIKIpathway annotation to determine pathways in which the proteins of interest were significantly over-represented. Results and conclusion: Autoantibody responses to 57 proteins were significantly altered in the COVID-19 Disc cohort compared to healthy controls (p ≤ 0.05). The Rep cohort had altered autoantibody responses against 26 proteins compared to non-COVID-19 ICU patients who served as controls. Both cohorts showed substantial similarities (r ² = 0.73) and exhibited higher autoantibody responses to numerous transcription factors, immunomodulatory proteins, and human disease markers. Analysis of the combined cohorts revealed elevated autoantibody responses against SPANXN4, STK25, ATF4, PRKD2, and CHMP3 proteins in COVID-19 patients. The sequences for SPANXN4 and STK25 were cross-validated using sequence alignment tools. ELISA and Western blot further verified the autoantigen-autoantibody response of SPANXN4. SPANXN4 is essential for spermiogenesis and male fertility, which may predict a potential role for this protein in COVID-19-associated male reproductive tract complications, and warrants further research.

Keywords: COVID-19; SPANXN4; Stk25; autoantibodies; immunoproteomics; male reproductive system.

Copyright © 2023 Schmidt, Abdesselem, Suhre, Vaikath, Sohail, Al-Nesf, Bensmail, Mashod, Sarwath, Bernhardt, Schaefer-Ramadan, Tan, Morris, Schenck, Price, Mohamed-Ali, Al-Maadheed, Arredouani, Decock, Blackburn, Choi and El-Agnaf.

Abstract

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