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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Research Article Autoimmunity

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 1

Metabolic parameters in neutrophils from people in the control group, patients with APS, patients with aPL-only, and patients with Thromb (aPL–).

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Metabolic parameters in neutrophils from people in the control group, pa...
(A) 2-NBDG fluorescence in people in the control group (n = 8) and patients with APS (n = 13) by flow cytometry; **P < 0.01 using t test. (B) Extracellular flux analysis of neutrophils using the glycolysis stress test. Glycolytic capacity was defined as the extracellular acidification rate measuring the maximal cellular utilization of glycolysis. Presented as fold change compared with controls; *P < 0.05, **P < 0.01 using 1-way ANOVA with Holm-Šidák’s multiple comparison test. n = 42 controls, 34 APS, 9 aPL-only, and 9 Thromb (aPL–). (C) L-Lactate in cell culture supernatant from controls (n = 11) and patients with APS (n = 17); *P < 0.05 using t test. (D) Glycolytic capacity of patients with APS with a history of microvascular disease (defined as having a history of diffuse alveolar hemorrhage, thrombotic microangiopathy, or catastrophic APS, n = 10) as compared with patients with APS without these features (n = 24); *P < 0.05 using t test. (E) Intracellular G6PD enzyme activity from controls (n = 11) and patients with APS (n = 17); *P < 0.05 using t test. (F) Total cellular ROS production in people in the control group (n = 8) and patients with APS (n = 13) as measured with DCFDA fluorescence by flow cytometry; **P < 0.01 using t test. (G) Glycogen stores in controls (n = 22) and patients with APS (n = 27); *P < 0.05 using t test.