Soluble CD13 is a potential mediator of neutrophil-induced thrombogenic inflammation in SARS-CoV-2 infection
Pei-Suen Tsou, et al.
Pei-Suen Tsou, et al.
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Research Article COVID-19 Immunology Infectious disease

Soluble CD13 is a potential mediator of neutrophil-induced thrombogenic inflammation in SARS-CoV-2 infection

Abstract

The soluble variant of the ectopeptidase CD13 (sCD13), released from the cell surface by matrix metalloproteinase 14 (MMP14), is a potent pro-inflammatory mediator, displaying chemotactic, angiogenic, and arthritogenic properties through bradykinin receptor B1 (B1R). We revealed a link between sCD13 and amplified neutrophil-mediated inflammatory responses in SARS-CoV-2 infection. sCD13 was markedly elevated in patients with COVID-19 and correlated with disease severity and variants, ethnicity, inflammation markers, and neutrophil extracellular trap formation (NETosis). Neutrophils treated with sCD13 showed heightened NETosis and chemotaxis, which were inhibited by sCD13 receptor blockade. Meanwhile sCD13 did not induce platelet aggregation. Single-cell analysis of COVID-19 lungs revealed coexpression of CD13 and MMP14 by various cell types, and higher CD13 expression compared with controls. Neutrophils with high CD13 mRNA were enriched for genes associated with immaturity, though CD13 protein expression was lower. Histological examination of COVID-19 lungs revealed CD13-positive leukocytes trapped in vessels with fibrin thrombi. Flow cytometry verified the presence of B1R and a second sCD13 receptor, protease-activated receptor 4, on monocytes and neutrophils. These findings identify sCD13 as a potential instigator of COVID-19–associated NETosis, potentiating vascular stress and thromboembolic complications. The potent pro-inflammatory effects of sCD13 may contribute to severe COVID-19, suggesting that sCD13 and its receptors might be therapeutic targets.

Authors

Pei-Suen Tsou, Ramadan A. Ali, Chenyang Lu, Gautam Sule, Carmelo Carmona-Rivera, Serena Lucotti, Yuzo Ikari, Qi Wu, Phillip L. Campbell, Mikel Gurrea-Rubio, Kohei Maeda, Sharon E. Fox, William D. Brodie, Megan N. Mattichak, Caroline Foster, Ajay Tambralli, Srilakshmi Yalavarthi, M. Asif Amin, Katarina Kmetova, Bruna Mazetto Fonseca, Emily Chong, Yu Zuo, Michael D. Maile, Luisa Imberti, Arnaldo Caruso, Francesca Caccuri, Virginia Quaresima, Alessandra Sottini, Douglas B. Kuhns, Danielle Fink, Riccardo Castagnoli, Ottavia M. Delmonte, Heather Kenney, Yu Zhang, Mary Magliocco, Helen Su, Luigi Notarangelo, Rachel L. Zemans, Yang Mao-Draayer, Irina R. Matei, Mirella Salvatore, David Lyden, Yogendra Kanthi, Mariana J. Kaplan, Jason S. Knight, David A. Fox

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Figure 4

Cells expressing CD13, MMP14, B1R, and PAR4 are present in lungs in patients with COVID-19, and cellular levels of CD13 mRNA correlate with disease severity and neutrophil maturity.

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Cells expressing CD13, MMP14, B1R, and PAR4 are present in lungs in pati...
Single-cell RNA-Seq results of nasopharyngeal/pharyngeal swabs, bronchial brushings and bronchial lavages were generated from patients with COVID-19 (n = 19) and healthy controls (n = 5). Data are extracted from Chua et al. (29). (A) Both ANPEP (codes for CD13) and MMP14 are expressed on various epithelial cells and macrophages. ANPEP is also expressed on neutrophils while MMP14 is expressed on mast cells. BDKRB1 (codes for B1R) is expressed on some epithelial cells with the highest expression on secretory cells and ciliated cells. F2RL3 (codes for PAR4) was barely detected in this dataset but seemed to be expressed by epithelial cells. Cell abbreviations are defined in Supplemental Figure 8. (B) Cellular ANPEP expression in nasopharyngeal cells is elevated in patients with COVID-19 (moderate COVID-19 n = 8, severe COVID-19 n = 11) compared with healthy controls (n = 5). (C) Using the median expression levels of ANPEP in patients, patients with COVID-19 were divided into 2 populations: ANPEP-high and ANPEP-low. Differentially expressed genes in neutrophils from these 2 groups were shown in the volcano plot (P < 1 × 10–20, |log2(fold-change)| < 0.25). (D) Pathway analysis of the differentially expressed genes in ANPEP-high and ANPEP-low neutrophils showed pathways related to neutrophil degranulation, leukocyte activation, and inflammation. (E) The neutrophils from ANPEP-high patients with COVID-19 (n = 10) showed a gene signature of immature-like neutrophils characterized by the overexpression of genes coding for several granule-content proteins (healthy controls n = 5, ANPEP-low n = 9). (F) The score for neutrophil immaturity was higher in critically ill patients with COVID-19 (n = 11) compared with moderate patients (left, n = 8). The median neutrophil-immaturity score of neutrophils in ANPEP-high patients (n = 10) was lower than that in ANPEP-low patients (right, n = 9). (G) The neutrophil-immaturity signature was most prominent in ANPEP-high patients (n = 10) as developing neutrophils versus mature neutrophils from the peripheral blood were examined (healthy controls n = 5, ANPEP-low n = 9). Results are expressed as mean ± SD. ****P < 0.0001. Significance was determined by 1-way ANOVA (B and G) and Mann-Whitney test (F).