MicroRNAs and Neutrophil Activation Markers Predict Venous Thrombosis in Pancreatic Ductal Adenocarcinoma and Distal Extrahepatic Cholangiocarcinoma

Abstract

Cancer-associated venous thrombosis (VTE) increases mortality and morbidity. However, limited tools are available to identify high risk patients. Upon activation, neutrophils release their content through different mechanisms, thereby prompting thrombosis. We explored plasma microRNAs (miRNAs) and neutrophil activation markers to predict VTE in pancreatic ductal adenocarcinoma (PDAC) and distal extrahepatic cholangiocarcinoma (DECC). Twenty-six PDAC and 6 DECC patients recruited at cancer diagnosis, were examined for deep vein thrombosis and pulmonary embolisms, and were then followed-up with clinical examinations, blood collections, and biCUS. Ten patients developed VTE and were compared with 22 age- and sex-matched controls. miRNA expression levels were measured at diagnosis and right before VTE, and neutrophil activation markers (cell-free DNA, nucleosomes, calprotectin, and myeloperoxidase) were measured in every sample obtained during follow-up. We obtained a profile of 7 miRNAs able to estimate the risk of future VTE at diagnosis (AUC = 0.95; 95% Confidence Interval (CI) (0.987, 1)) with targets involved in the pancreatic cancer and complement and coagulation cascades pathways. Seven miRNAs were up- or down-regulated before VTE compared with diagnosis. We obtained a predictive model of VTE with calprotectin as predictor (AUC = 0.77; 95% CI (0.57, 0.95)). This is the first study that addresses the ability of plasma miRNAs and neutrophil activation markers to predict VTE in PDAC and DECC.

Increase of Neutrophil Activation Markers in Venous Thrombosis—Contribution of Circulating Activated Protein C

“…The causes of thrombosis are both genetic and acquired; however, despite the large increase in the number of identified risk factors, still a substantial number of thrombotic events occur spontaneously without an apparent origin, which may reflect causes not yet discovered. Provided the interplay between immunity and thrombosis, where neutrophils play a predominant role; markers of neutrophil activation may allow identifying patients with high thrombotic risk, as we previously demonstrated in cancer patients [33,34]. Thus, we explored the relation of neutrophil activation markers and the risk of thrombosis.…”

“…Following the strategy addressed in previous studies [25][26][27][28][29]33,34,54,55], we evaluated whether the different markers of neutrophil activation measured in plasma had the same cellular origin by means of their correlation. We found a significant correlation between levels of cfDNA and calprotectin (r = 0.28; p < 0.0001), similar to that observed in previous plasma studies [27,33], and also between neutrophil count and calprotectin (Spearman r = 0.18; p = 0.003) indicating that they might have, to some extent, the same origin, probably an increased activation of neutrophils or NETosis. Additionally, the inflammatory marker NLR has been previously ascertained as a simple and valuable marker for VTE prognosis [56][57][58][59] and also for other thrombotic disorders [60][61][62][63].…”

“…Previous studies carried out to clarify the role of NETs in thrombosis have been mainly conducted in animal models [2,4,6,20,21]. However, there is already incipient evidence about the relationship between NETs and the risk of thrombosis in patients with various prothrombotic and inflammatory disorders [22][23][24][25][26][27][28][29][30][31][32][33][34]. In the particular scenario of VTE, in a case-control study with a very small number of samples, Díaz et al [26] observed a significant increase in plasma cell-free DNA (cfDNA) concentration in patients with acute DVT.…”

From Classical Laboratory Parameters to Novel Biomarkers for the Diagnosis of Venous Thrombosis

“…Other studies suggest that microparticles carry both nuclear and cytoplasmic ribonucleic acid (RNA) [83] and deoxyribonucleic acid (DNA) [84], especially microRNA. Additionally, microparticles have an important role in transferring microRNA from endothelial progenitor cells to endothelial cells, and this horizontal transfer of genetic mass between platelets and endothelial cells may influence thrombosis [14].…”

Role of microRNAs in Venous Thromboembolism

“…Even though several risk stratification tools have been proposed to identify a subset of cancer patients at high risk of developing VTE, their clinical usefulness remains under debate [56]. It is of interest that in a recent study, Oto et al (2020) [57] identified a profile consisting of 7 miRNAs (miR-486-5p, miR-106b-5p, let-7i-5p, let-7g-5p, miR-144-3p, miR-19a-3p, and miR-103a-3p) associated with the occurrence of a future VTE event during follow-up in a cohort of 32 patients with pancreatic ductal adenocarcinoma and distal extrahepatic cholangiocarcinoma.…”