Neutrophil glucose flux as a therapeutic target in antiphospholipid syndrome
Ajay Tambralli, … , Costas A. Lyssiotis, Jason S. Knight
Ajay Tambralli, … , Costas A. Lyssiotis, Jason S. Knight
Published June 13, 2024
Citation Information: J Clin Invest. 2024;134(15):e169893. https://doi.org/10.1172/JCI169893.
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Neutrophil glucose flux as a therapeutic target in antiphospholipid syndrome

Abstract

Neutrophil hyperactivity and neutrophil extracellular trap release (NETosis) appear to play important roles in the pathogenesis of the thromboinflammatory autoimmune disease known as antiphospholipid syndrome (APS). The understanding of neutrophil metabolism has advanced tremendously in the past decade, and accumulating evidence suggests that a variety of metabolic pathways guide neutrophil activities in health and disease. Our previous work characterizing the transcriptome of APS neutrophils revealed that genes related to glycolysis, glycogenolysis, and the pentose phosphate pathway (PPP) were significantly upregulated. Here, we found that neutrophils from patients with APS used glycolysis more avidly than neutrophils from people in the healthy control group, especially when the neutrophils were from patients with APS with a history of microvascular disease. In vitro, inhibiting either glycolysis or the PPP tempered phorbol myristate acetate– and APS IgG–induced NETosis, but not NETosis triggered by a calcium ionophore. In mice, inhibiting either glycolysis or the PPP reduced neutrophil reactive oxygen species production and suppressed APS IgG–induced NETosis ex vivo. When APS-associated thrombosis was evaluated in mice, inhibiting either glycolysis or the PPP markedly suppressed thrombosis and circulating NET remnants. In summary, these data identify a potential role for restraining neutrophil glucose flux in the treatment of APS.

Authors

Ajay Tambralli, Alyssa Harbaugh, Somanathapura K. NaveenKumar, Megan D. Radyk, Christine E. Rysenga, Kaitlyn Sabb, Julia M. Hurley, Gautam J. Sule, Srilakshmi Yalavarthi, Shanea K. Estes, Claire K. Hoy, Tristin Smith, Cyrus Sarosh, Jacqueline A. Madison, Jordan K. Schaefer, Suman L. Sood, Yu Zuo, Amr H. Sawalha, Costas A. Lyssiotis, Jason S. Knight

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Figure 2

Glycolysis and the PPP are required for APS IgG–induced human neutrophil NETosis and ROS production.

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Glycolysis and the PPP are required for APS IgG–induced human neutrophil...
(A) Using a metabolic flux analyzer, neutrophils from controls were treated with the indicated stimuli, and ECAR (left) and OCR (right) trends were measured over 4 hours. These data are representative of 3 independent experiments. (B) Neutrophils from people in the control group (n = 5) were treated with PBS, 2-DG (10 mM), G6PDi-1 (50 μM), or DPI (10 μM) for 30 minutes and stimulated with total IgG fractions prepared from people in the control group (control IgG, 10 μg/mL), total IgG fractions prepared from patients with APS (APS IgG, 10 μg/mL), PMA (40 nM), or Ca iono (10 μM) for 3 hours and NETosis was quantified using SYTOX Green. All data are presented as fold change compared with neutrophils that were not treated with any inhibitors or stimuli. (C) Neutrophils from people in the control group (n = 3) were treated with inhibitors as in B and then stimulated as indicated for 1 hour. Cytosolic ROS production was quantified using the Amplex Red reagent. For B–C, # represents the effectiveness of NETosis induction or ROS production for APS IgG, PMA, and Ca iono compared with control IgG using 1-way ANOVA with Holm-Šidák’s multiple comparison test; #P < 0.05, ####P < 0.0001 * represents the change in NETosis or ROS production with the inhibitors in each stimulant group using 1-way ANOVA with Holm-Šidák’s multiple comparison test; **P < 0.01, ***P < 0.001, ****P < 0.0001.