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. 2019 Nov 26;8(12):2072.
doi: 10.3390/jcm8122072.

Interleukin-1β Mediates Arterial Thrombus Formation via NET-Associated Tissue Factor

Luca Liberale  1   2 , Erik W Holy  3 , Alexander Akhmedov  1 , Nicole R Bonetti  1   4 , Fabian Nietlispach  3 , Christian M Matter  1   3 , François Mach  5 , Fabrizio Montecucco  2   6 , Jürg H Beer  1   4 , Francesco Paneni  1   3   7 , Frank Ruschitzka  3 , Peter Libby  8 , Thomas F Lüscher  1   9 , Giovanni G Camici  1   3   7
Affiliations

Interleukin-1β Mediates Arterial Thrombus Formation via NET-Associated Tissue Factor

Luca Liberale et al. J Clin Med. .

Abstract

CANTOS reported reduced secondary atherothrombotic events in patients with residual inflammatory risk treated with the inhibitory anti-IL-1β antibody, Canakinumab. Yet, mechanisms that underlie this benefit remain elusive. Recent work has implicated formation of neutrophil extracellular traps (NETosis) in arterial thrombosis. Hence, the present study explored the potential link between IL-1β, NETs, and tissue factor (TF)-the key trigger of the coagulation cascade-in atherothrombosis. To this end, ST-elevation myocardial infarction (STEMI) patients from the Swiss multicenter trial SPUM-ACS were retrospectively and randomly selected based on their CRP levels. In particular, 33 patients with STEMI and high C-reactive protein (CRP) levels (≥ 10 mg/L) and, 33 with STEMI and low CRP levels (≤ 4 mg/L) were investigated. High CRP patients displayed elevated circulating IL-1β, NETosis, and NET-associated TF plasma levels compared with low CRP ones. Additionally, analysis of patients stratified by circulating IL-1β levels yielded similar results. Moreover, NETosis and NET-associated TF plasma levels correlated positively in the whole population. In addition to the above, translational research experiments provided mechanistic confirmation for the clinical data identifying IL-1β as the initial trigger for the release of the pro-coagulant, NET-associated TF. In conclusion, blunted TF presentation by activated neutrophils undergoing NETosis may provide a mechanistic explanation to reduced secondary atherothrombotic events as observed in canakinumab-treated patients in CANTOS.

Keywords: Canakinumab; IL-1β; arterial thrombosis; neutrophil extracellular traps; tissue factor.

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Conflict of interest statement

T.F.L. and C.M.M. have been member of the Canakinumab advisory board of Novartis and have received honoraria as well as educational grants to the institution. P.L. is an unpaid consultant to, or involved in clinical trials for Amgen, AstraZeneca, Esperion Therapeutics, Ionis Pharmaceuticals, Kowa Pharmaceuticals, Novartis, Pfizer, Sanofi-Regeneron, and XBiotech, Inc. PL is a member of scientific advisory board for Amgen, Corvidia Therapeutics, DalCor Pharmaceuticals, IFM Therapeutics, Kowa Pharmaceuticals, Olatec Therapeutics, Medimmune, Novartis, and XBiotech, Inc. Libby’s laboratory has received research funding in the last 2 years from Novartis. All other authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Levels of IL-1β, neutrophil extracellular traps (NETosis), NET-associated tissue factor and their relationships in patients with coronary disease. (A) STEMI patients with high systemic inflammation have higher IL-1β plasma levels than those with lower C-reactive protein (CRP) levels (n = 33). (B) Patients with STEMI and high CRP levels showed increased levels of NETosis assessed by plasma MPO-DNA complexes (n = 32–33) (C) Plasma tissue factor TF)-DNA complexes rose in patients with STEMI and high circulating CRP compared to less inflamed patients (n = 33). (D–E) Similarly, in patients categorized according to circulating IL-1β levels (above or below the median value of our cohort 0.1041 pg/mL), those with high levels of this cytokine also showed increased myeloperoxidase (MPO)-DNA and TF–DNA circulating complexes (n = 33) (F) A positive relationship was detectable among plasma IL-1β and MPO-DNA complexes in the whole cohort (n = 66). **p < 0.01. CRP = C reactive protein, IL-1β = interleukin-1β, MPO = myeloperoxidase, STEMI = ST-elevated myocardial infarction, TF = tissue factor.
Figure 1
Figure 1
Levels of IL-1β, neutrophil extracellular traps (NETosis), NET-associated tissue factor and their relationships in patients with coronary disease. (A) STEMI patients with high systemic inflammation have higher IL-1β plasma levels than those with lower C-reactive protein (CRP) levels (n = 33). (B) Patients with STEMI and high CRP levels showed increased levels of NETosis assessed by plasma MPO-DNA complexes (n = 32–33) (C) Plasma tissue factor TF)-DNA complexes rose in patients with STEMI and high circulating CRP compared to less inflamed patients (n = 33). (D–E) Similarly, in patients categorized according to circulating IL-1β levels (above or below the median value of our cohort 0.1041 pg/mL), those with high levels of this cytokine also showed increased myeloperoxidase (MPO)-DNA and TF–DNA circulating complexes (n = 33) (F) A positive relationship was detectable among plasma IL-1β and MPO-DNA complexes in the whole cohort (n = 66). **p < 0.01. CRP = C reactive protein, IL-1β = interleukin-1β, MPO = myeloperoxidase, STEMI = ST-elevated myocardial infarction, TF = tissue factor.
Figure 2
Figure 2
Anti-IL-1β antibody retards arterial thrombosis. (A) In an attempt to simulate the design of Canakinumab Anti-inflammatory Thrombosis Outcomes Study (CANTOS), which enrolled patients with residual inflammatory risk, animals received lipopolysaccharide (LPS) (5 μg/g, i.p) 10 h before undergoing photochemically induced carotid thrombosis. The anti-mouse IL-1β monoclonal antibody was administered to the animal intravenously via tail vein injection 5 h before thrombosis at a single dose of 10 µg/g, vehicle (i.e., NaCl 0.9%) was used as negative control. (B) Animals treated with monoclonal antibody against IL-1β (anti IL-1β Ab) showed increased time to occlusion as compared to vehicle-treated ones (n = 7–8). (C) Representative trace of mean blood flow until occlusion (mean flow ≤ 0.1 mL for 1 min) in the two study groups. (D,E) Baseline blood flow and heart rate did not differ among treated and untreated animals (n = 7–8). ***p < 0.001. IL-1β = interleukin-1β, LPS = lipopolysaccharide.
Figure 3
Figure 3
IL-1β blockade reduces different tissue factor pools in mice with arterial thrombosis. (A) Treatment with the canakinumab-surrogate antibody (anti IL-1β Ab) reduced plasma TF levels as assessed by ELISA (n = 7–8). (B) Treated and untreated animals showed no difference in terms of TF concentration in aorta lysates (n = 7). * p < 0.05. ELISA = enzyme-linked immunosorbent assay, IL-1β = interleukin-1β, TF = tissue factor.
Figure 4
Figure 4
IL-1β blockade limits NETosis and NET-associate tissue factor levels. (A) Animal treated with the murine anti IL-1β antibody showed a significant reduction in the level of NETosis plasma marker MPO-DNA complexes (n = 7) (B) Levels of NET-associated TF fell after thrombosis in animals treated with the anti-IL-1β antibody as assessed by plasma TF–DNA complexes (n = 7). * p < 0.05. IL-1β = interleukin-1β, MPO = myeloperoxidase, NET = neutrophil extracellular trap, TF = tissue factor.
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References

    1. Ridker P.M., Everett B.M., Thuren T., MacFadyen J.G., Chang W.H., Ballantyne C., Fonseca F., Nicolau J., Koenig W., Anker S.D., et al. Antiinflammatory Therapy with Canakinumab for Atherosclerotic Disease. N. Engl. J. Med. 2017;377:1119–1131. doi: 10.1056/NEJMoa1707914. - DOI - PubMed
    1. Libby P. Inflammation in atherosclerosis. Nature. 2002;420:868–874. doi: 10.1038/nature01323. - DOI - PubMed
    1. Lippi G., Franchini M., Targher G. Arterial thrombus formation in cardiovascular disease. Nat. Rev. Cardiol. 2011;8:502–512. doi: 10.1038/nrcardio.2011.91. - DOI - PubMed
    1. Montecucco F., Liberale L., Bonaventura A., Vecchie A., Dallegri F., Carbone F. The Role of Inflammation in Cardiovascular Outcome. Curr. Atheroscler. Rep. 2017;19:11. doi: 10.1007/s11883-017-0646-1. - DOI - PubMed
    1. Hansson G.K., Libby P., Tabas I. Inflammation and plaque vulnerability. J. Intern. Med. 2015;278:483–493. doi: 10.1111/joim.12406. - DOI - PMC - PubMed