Glucose associated NETosis in patients with ST-elevation myocardial infarction: an observational study

Ragnhild Helseth, Eva Cecilie Knudsen, Jan Eritsland, Trine Baur Opstad, Harald Arnesen, Geir Øystein Andersen, Ingebjørg Seljeflot, Ragnhild Helseth, Eva Cecilie Knudsen, Jan Eritsland, Trine Baur Opstad, Harald Arnesen, Geir Øystein Andersen, Ingebjørg Seljeflot

Abstract

Background: Neutrophil extracellular traps (NETs) have recently been identified as mediators in atherothrombosis. Although NETosis in general has been suggested to be glucose dependent, the transferability to patients with acute ST-elevation myocardial infarction (STEMI) is unclear. We assessed whether the NETs markers double-stranded deoxyribonucleid acid (dsDNA) and myeloperoxidase-DNA (MPO-DNA) associated with plasma glucose and the glucometabolic status in the acute phase and 3 months after a STEMI. We also explored whether an acute glucose load resulted in upregulated NETosis by assessment of peptidylarginine deiminase 4 (PAD4) gene expression.

Methods: In total, 224 STEMI patients were prospectively enrolled and underwent blood sampling acutely (median 16.5 h after PCI) and after 3 months. Glucometabolic status was defined based on the results of an oral glucose tolerance test (OGTT) as normal glucose regulation (NGR), impaired fasting glucose (IFG), impaired glucose tolerance (IGT) or type 2 diabetes (T2DM). dsDNA and MPO-DNA were measured in serum, while PAD4 mRNA was measured in circulating leukocytes by RT-PCR.

Results: dsDNA levels were significantly correlated to plasma glucose both acutely and after 3 months (r = 0.12 and r = 0.17, both p < 0.02), whereas MPO-DNA was not. No associations with the glucometabolic status were encountered for dsDNA and MPO-DNA acutely, but after 3 months dsDNA levels were elevated in patients with IFG and T2DM vs. NGR (428 vs. 371 ng/ml and 408 vs. 371 ng/ml, both p < 0.045). During the acute glucose load after 3 months, dsDNA and MPO-DNA remained unchanged while PAD4 mRNA increased significantly (RQ 0.836 vs. 0.920, p = 0.02).

Conclusions: In this cohort of STEMI patients, levels of dsDNA associated with plasma glucose both in the acute and stable condition. The glucometabolic status was not substantially related to the selected NETs markers, however, an acute glucose load by OGTT performed after 3 months resulted in increased PAD4 expression, suggestive of enhanced NETosis in the aftermath of STEMI.

Trial registration: www.clinicaltrials.gov, NCT00926133 . Registered June 23, 2009.

Keywords: Acute ST-elevation myocardial infarction (STEMI); Double-stranded deoxyribonucleid acid (dsDNA); Glucometabolic status; Glucose; Immunothrombosis; Innate immunity; Neutrophil activation; Neutrophil extracellular traps (NETs).

Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
The time profiles of NETs markers and change during OGTT. a dsDNA, b MPO-DNA, c PAD4 mRNA. Values are given as medians (25, 75 percentiles). p values are based on the Wilcoxon Signed Rank test. *: Significant change from the acute phase. **: Significant change during the course of OGTT. dsDNA: double-stranded deoxyribonucleic acid. MPO-DNA: myeloperodidase deoxyribonucleid acid. OGTT: oral glucose tolerance test. OD: optical density units. PAD4 mRNA: peptidyl arginine deiminase 4 messenger ribonucleic acid
Fig. 2
Fig. 2
Correlation between glucose and dsDNA. a Acute phase, b at 3 months. p values are based on the Spearman’s rho. dsDNA: double-stranded deoxyribonucleic acid. OGTT: oral glucose tolerance test
Fig. 3
Fig. 3
NETs markers related to the glucometabolic status at 3 months. Values are given as medians (25, 75 percentiles). a-c: dsDNA levels in NGR vs. IFG, IGT and T2DM, respectively. d-f: MPO-DNA levels in NGR vs. IFG, IGT and T2DM, respectively. g-i: PAD4 mRNA levels in NGR vs. IFG, IGT and T2DM, respectively.p values are based on the Mann-Whitney U test. dsDNA: double-stranded deoxyribonucleic acid. IFG: impaired fasting glucose. IGT: impaired glucose tolerance. MPO-DNA: myeloperoxidase deoxyribonucleic acid. NGR: normal glucose regulation. OD: optical density units. PAD4 mRNA: peptidylarginine deiminase 4 messenger ribonucleic acid. RQ: relative quantification values. T2DM: type 2 diabetes mellitus
Fig. 4
Fig. 4
Suggested metabolic pathways involved in glucose mediated NETosis, simplified schematic illustration

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