ENTPD-1 disrupts inflammasome IL-1β–driven venous thrombosis
Vinita Yadav, … , David J. Pinsky, Yogendra Kanthi
Vinita Yadav, … , David J. Pinsky, Yogendra Kanthi
Published April 16, 2019
Citation Information: J Clin Invest. 2019;129(7):2872-2877. https://doi.org/10.1172/JCI124804.
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Concise Communication Inflammation Vascular biology

ENTPD-1 disrupts inflammasome IL-1β–driven venous thrombosis

Abstract

Deep vein thrombosis (DVT), caused by alterations in venous homeostasis, is the third most common cause of cardiovascular mortality, however, key molecular determinants in venous thrombosis have not been fully elucidated. Several lines of evidence indicate that DVT occurs at the intersection of dysregulated inflammation and coagulation. The enzyme ectonucleoside tri(di)phosphohydrolase (ENTPD1, also known as CD39) is a vascular ecto-apyrase on the surface of leukocytes and the endothelium that inhibits intravascular inflammation and thrombosis by hydrolysis of phosphodiester bonds from nucleotides released by activated cells. Here, we evaluated the contribution of CD39 to venous thrombosis in a restricted-flow model of murine inferior vena cava stenosis. CD39 deficiency conferred a greater than 2-fold increase in venous thrombogenesis, characterized by increased leukocyte engagement, neutrophil extracellular trap formation, fibrin, and local activation of tissue factor in the thrombotic milieu. This venous thrombogenesis was orchestrated by increased phosphorylation of the p65 subunit of NF-κB, activation of the NLR family pyrin domain–containing 3 (NLRP3) inflammasome, and IL-1β release in CD39-deficient mice. Substantiating these findings, an IL-1β–neutralizing antibody or the IL-1 receptor inhibitor anakinra attenuated the thrombosis risk in CD39-deficient mice. These data demonstrate that IL-1β is a key accelerant of venous thrombo-inflammation, which can be suppressed by CD39. CD39 inhibits in vivo crosstalk between inflammation and coagulation pathways and is a critical vascular checkpoint in venous thrombosis.

Authors

Vinita Yadav, Liguo Chi, Raymond Zhao, Benjamin E. Tourdot, Srilakshmi Yalavarthi, Benjamin N. Jacobs, Alison Banka, Hui Liao, Sharon Koonse, Anuli C. Anyanwu, Scott H. Visovatti, Michael A. Holinstat, J. Michelle Kahlenberg, Jason S. Knight, David J. Pinsky, Yogendra Kanthi

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Figure 2

CD39 tempers thrombus cellularity and leukocyte recruitment to growing thrombus.

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CD39 tempers thrombus cellularity and leukocyte recruitment to growing t...
(A) Neutrophil-stained (Ly6G) venous thrombi (brown, Ly6G; n = 5 for each). Scale bars: 1 mm and 50 μm (insets). (B) Immunoblot for Ly6G in venous thrombi from WT and Cd39+/– mice (n = 5 for each). (C) H3-cit expression indicates NET formation by immunofluorescence microscopy following ex vivo stimulation with 100 nM PMA (green, H3-cit; blue, DNA; white arrowheads, NETs). WT n = 4, and Cd39+/– n = 3. Scale bars: 100 μm. Representative images are shown. (D) H3-cit content in venous thrombus lysates (n = 4 for each). (E) Venous thrombus weight and extension in WT mice following adoptive transfer of WT or Cd39+/– neutrophils (n = 15 for each). Data represent the mean ± SEM. *P < 0.05 and **P < 0.01, by 2-tailed Student’s t test.