2016;115(3):332–8. Cancer Res. Data are expressed as mean ± standard deviation. Cancer Gene Ther. 2015;110(3):20. Correspondence to Tumor mice had an elevated coagulation index compared with sham controls, suggestive of hypercoagulability (Fig. 5a). Diaz JA, Fuchs TA, Jackson TO, Kremer Hovinga JA, Lammle B, Henke PK, et al. Supernatant was collected after 4 h and the formation of NETs was confirmed by measuring supernatant DNA using Quant-iT Picogreen (Invitrogen, Grand Island, NY, USA, MP07581) and by fluorescence microscopy to visualize NET formation using DNA staining with Hoechst (Additional file 1: Figure S1). Autophagy, a cancer cell survival mechanism whereby damaged organelles, proteins and other intracellular components are recycled, appears to be critical for NET formation in pancreatic cancer [13]. He obtained complete remission and he is still alive and well after a treatment with gemcitabine and capecitabine, plus IV Paricalcitol (25 mcg 3x’s/week) and hydroxychloroquine (600 mg BID) http://apc.amegroups.com/article/view/4269/5197 The hypercoagulable state associated with pancreatic adenocarcinoma (PDA) results in increased risk of venous thromboembolism, leading to substantial morbidity and mortality. The interaction between NETs and platelets has been implicated in the pathogenesis of deep vein thrombosis [21]. 2016;76(6):1367–80. ], Overall Survival [ Time Frame: All patients were followed until death. Analysis was performed by using Student’s two tailed t-test or 1-way ANOVA with Tukey’s post-hoc test using Graph Pad Prism software (GraphPad, San Diego CA, USA). Abdol Razak N, Elaskalani O, Metharom P. Pancreatic Cancer-induced neutrophil extracellular traps: a potential contributor to Cancer-associated thrombosis. Blue = WT, Red = RAGE KO, Circle = Sham, Triangle = Tumor. There was no correlation between plasma DNA and VTE in HCQ treated patients. Experimental: Hydroxychloroquine 400 mg b.i.d. A 10 fold dilution was performed and mouse tissue factor levels were measured using the F3 / CD142 / Tissue factor ELISA per the manufacturer’s instruction (LS Bio, LS-F14709, Seattle, WA, USA). PubMed  Oncoimmunology. Front Immunol. Objective: To investigate whether hydroxychloroquine treatment is associated with major adverse cardiovascular events (MACE) (myocardial infarction, ischemic stroke, or cardiovascular-associated death) in patients with cutaneous LE (CLE) or systemic LE (SLE). However, because CQ also has direct antiplatelet effects, it is difficult to completely attribute all its effects to inhibition of NETosis. Keywords provided by Brian Wolpin, MD, MPH, Dana-Farber Cancer Institute: Why Should I Register and Submit Results? Google ScholarÂ. Google ScholarÂ. Hypercoagulable changes are detectable on rotational thromboelastometry, similar to TEG, in patients with abdominal malignancy [50]. (DOCX 14 kb). Tumor burdened mice had heightened platelet activation compared to sham controls (A). Safety and biologic response of pre-operative autophagy inhibition in combination with gemcitabine in patients with pancreatic adenocarcinoma. Recently, neutrophil extracellular traps (NETs), whereby activated neutrophils release their intracellular contents containing DNA, histones, tissue factor, high mobility group box 1 (HMGB1) and other components have been implicated in PDA and in cancer-associated thrombosis. This phase I trial studies the best dose of hydroxychloroquine when given together with binimetinib in treating patients with KRAS gene mutated pancreatic cancer that has spread to other places in the body (metastatic). engineered models of pancreatic cancer (1). Hydroxychloroquine is approved for the treatment of non-cancerous illnesses such as rheumatoid arthritis and systemic lupus erythematous. To study the role of chloroquine inhibition of NETs and hypercoagulability using a more informative and clinically translatable approach, we utilized thromboelastograms to evaluate whether treatment with chloroquine decreases hypercoagulability in orthotopic murine pancreatic cancer. Espinola RG, Pierangeli SS, Gharavi AE, Harris EN. Neutrophil extracellular traps promote thrombin generation through platelet-dependent and platelet-independent mechanisms. Nosal R, Jancinova V, Danihelova E. Chloroquine: a multipotent inhibitor of human platelets in vitro. J Exp Med. The team uncovered evidence to suggest that CQ and HCQ could be effective for the treatment of a number of cancers, including glioblastoma — … CQ inhibition of NETs reverses platelet aggregation and decreases tissue factor. Following standard IHC deparaffinization protocol, sections were subject to antigen retrieval using 10 mM Citric acid buffer. Women of child-bearing potential and men must agree to use adequate contraception prior to study entry and for the duration of study participation. Importantly, treatment with CQ in PAD4 KO mice, incapable of forming NETs, had minimal effect, suggesting that CQ decreases platelet aggregation through inhibition of NETs. Resected pancreatic specimens from patients with pancreatic adenocarcinoma were stained and imaged using the following protocol. Curve analysis was performed using Haemonetics TEG software (version 4.2.3) and the R, K, angle, and MA were measured. Google ScholarÂ. The generation of these mice from a C57/Bl6 background has been previously described [16]. PubMed  Thromboelastogram (TEG) values for orthotopic tumor and sham mice with and without chloroquine (CQ) treatment, demonstrating that tumor mice have hypercoagulable elevations in K, angle, maximum amplitude (MA) and coagulation index (CI) compared with sham controls and that CQ reverses hypercoagulability as assessed by the CI. Kambas K, Mitroulis I, Ritis K. The emerging role of neutrophils in thrombosis-the journey of TF through NETs. Paricalcitol (an analog of vitamin D) is also available orally if you can’t get it IV, but I don’t think it works as well. Here is some info that you might find interesting: Pre and post-treatment results were compared using paired t-test. Manage cookies/Do not sell my data we use in the preference centre. Binimetinib may stop the growth of tumor cells … Furthermore, RAGE KO mice, which have diminished NET formation, also had lower levels of serum tissue factor (Fig. 3b). CQ reverses hypercoagulability in tumor burdened mice. Cancer Res. PAD4 and RAGE knockout mice, deficient in NET formation, were used to study the role of NETs in platelet aggregation, release of tissue factor and hypercoagulability. Arch Intern Med. Article  2006;166(4):458–64. Cancers predispose neutrophils to release extracellular DNA traps that contribute to cancer-associated thrombosis. CAS  We confirmed these observations and expanded on this mechanism to include the receptor for advanced glycation end products (RAGE), a known receptor for extracellular DNA, as a critical component of NET mediated platelet aggregation in pancreatic cancer. 2009;27(Suppl 1):63–74. We next assessed the rate of venous thromboembolism (VTE) in patients treated with pre-operative hydroxychloroquine as part of two separate clinical trial protocols. Front Immunol. Incidence, outcome and risk stratification tools for venous thromboembolism in advanced pancreatic cancer - a retrospective cohort study. *p < 0.05. At the onset of the COVID-19 pandemic, hydroxychloroquine became a hot topic as a possible treatment for the virus.Clinical trials largely found that the drug was not a viable treatment option. Doring Y, Soehnlein O, Weber C. Neutrophil extracellular traps in atherosclerosis and Atherothrombosis. J Clin Oncol. Cancer Investig. Neutrophil histone modification by peptidylarginine deiminase 4 is critical for deep vein thrombosis in mice. CR or PR confirmation is required >/= 4 weeks. PAD4 KO mice are unable to form NETs as a result of genetic deficiency in protein arginine deiminase 4, an enzyme critical for NET formation that citrullinates histones to allow for DNA unwinding and expulsion from the cell [22]. 2005;44(3):293–8. The current work explores upregulation of platelet function and release of tissue factor as two mechanisms through which NETs contribute to hypercoagulability and thrombosis in pancreatic cancer. About the collective. Treatment with NET supernatant induced platelet aggregation in both human (Fig. 1b) and murine (Fig. 1c) blood in a dose dependent fashion and increased platelet activation (Additional file 2: Figure S2B). Neutrophil extracellular traps sequester circulating tumor cells and promote metastasis. In the current study, inhibition of NETs with chloroquine resulted in decreased platelet aggregation and lower levels of circulating tissue factor. These studies are designed to fail. Proc Natl Acad Sci U S A. BMC Cancer Novel pathways and therapeutic approaches to prevent VTE events are needed [6]. Proc Natl Acad Sci U S A. Pancreatic cancer is associated with a hypercoagulable state resulting in a high risk of venous thromboembolism (VTE), which affects up to 40% of patients during their course of disease [1,2,3]. These findings implicate a role for DNA and RAGE in NET induced platelet aggregation. Hydroxychloroquine is a relatively inexpensive drug currently available for the treatment of malaria and autoimmune diseases. P-values < 0.05 were considered statistically significant. Serum was collected after blood was allowed to clot for 30 min and then spun at 1000 g for 10 min. This phase I trial studies the sides effects and best dose of hydroxychloroquine when given together with trametinib in treating patients with pancreatic cancer that has spread to nearby tissue, lymph nodes or other places in the body and cannot be removed by surgery. 2014 Jun;19(6):637-8. doi: 10.1634/theoncologist.2014-0086. My treatment included chemo, plus IV Paricalcitol and hydroxychloroquine). Of note, the lone patient who developed VTE was treated as part of the dose escalation at 800 mg per day rather than at the maximum dose of 1200 mg. 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