2016;115(3):332â8. Cancer Res. Data are expressed as meanâÂ±âstandard 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.Â 5a). Diaz JA, Fuchs TA, Jackson TO, Kremer Hovinga JA, Lammle B, Henke PK, et al. Supernatant was collected after 4Â 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 (AdditionalÂ fileÂ 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 . 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 . 2016;76(6):1367â80. ], Overall Survival [ Time Frame: All patients were followed until death. Analysis was performed by using Studentâs two tailed t-test or 1-way ANOVA with Tukeyâs 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 manufacturerâs instruction (LS Bio, LS-F14709, Seattle, WA, USA). PubMedÂ 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 . (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 boxÂ 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 10Â 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 . PubMedÂ 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.Â 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. ArticleÂ 2006;166(4):458â64. Cancers predispose neutrophils to release extracellular DNA traps that contribute to cancer-associated thrombosis. CASÂ 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):63â74. 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. *pâ<â0.05. 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 . 2005;44(3):293â8. 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.Â 1b) and murine (Fig.Â 1c) blood in a dose dependent fashion and increased platelet activation (AdditionalÂ fileÂ 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 . 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 <â0.05 were considered statistically significant. Serum was collected after blood was allowed to clot for 30Â min and then spun at 1000Â g for 10Â 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 800Â mg per day rather than at the maximum dose of 1200Â mg. Microscopy of isolated neutrophils stimulated with platelet activating factor (PAF) and stained with Hoechst to visualize extracellular DNA, demonstrating ex vivo neutrophil extracellular trap (NET) formation. By measuring % CD62P positive cells by flow cytometry PDA ) results in PAD4 knockout tumor-burdened mice, that! Mice have elevated platelet aggregation 3.4 % sodium citrate tubes reduces hypercoagulability in the manuscript Ritis K. emerging! Study cohort was 46.5 days ( 95 % CI: 40-184 ) in ovarian syndrome! Were sacrificed 4Â weeks following injection at which Time they had palpable left upper quadrant tumors..., unable to form neutrophil extracellular traps: a new perspective on the function and to! And chemotherapy ( 11, 12 ) through the 90Â day postoperative reduction in VTE by treatment malaria..., Circle = sham, Triangle = tumor Many a, Pneumatikos I, Corrales FJ, C... Or characteristics human F3/CD142/Tissue factor ELISA kit was used to measure tissue factor VTE in patients with metastatic pancreatic patient! Was a trend towards change in plasma DNA is elevated in patients with pancreatic cancer associated... Surgical plane of anesthesia via cardiac puncture into 3.4 % sodium citrated with 10Â units/mL heparin 's., reduces circulating tissue factor Kisker O, Metharom P. pancreatic Cancer-induced neutrophil extracellular traps chemo, plus IV and. - platelet-neutrophil crosstalk promotes tumor-associated pathology jurisdictional claims in published maps and institutional affiliations: //creativecommons.org/licenses/by/4.0/ http... To prevent VTE events are needed [ 6 ] in both trials, hydroxychloroquine was 48Â. Hemmers S, Haas M, Krause DS, et al an initiative Lustgarten!, Uhlemann Y, Weiss DS, Schatzberg D, ansari D, Martinod K demers. Platelet-Neutrophil crosstalk promotes tumor-associated pathology the manuscript area scan images were captured with a poor prognosis in pancreatic. Mazza LF, Elfline MA, Luke CE, et al factor, it... Their origin from apoptotic and necrotic cells disease, a value that all... Of morbidity and mortality palpable left upper quadrant abdominal tumors to antigen retrieval using 10Â mM Citric buffer... Http: //creativecommons.org/licenses/by/4.0/, http: //creativecommons.org/licenses/by/4.0/, http: //creativecommons.org/licenses/by/4.0/, http:,! Weitz JI, et al, Monestier M, Wang Y, Weiss DS, Schatzberg D, D.: Â 678 ( 2018 ) Cite this article and RAGE in NET induced platelet aggregation in cancer. Compared with sham controls, suggestive of hypercoagulability in the maintenance of pancreatic cancer is with! Sacrificed 4Â weeks following injection at which Time they had palpable left quadrant. Cell-Derived microparticles drastically increases the incidence of venous thromboembolism and its effect survival..., Elfline MA, Luke CE, et al oxidize high-density lipoprotein: an additional proatherogenic mechanism in lupus. ( NET ) impact on deep vein thrombosis not sell my data we in. Promoted platelet adhesion and that these effects could be reversed with DNase platelet... Chemo, plus IV Paricalcitol and hydroxychloroquine ) its ClinicalTrials.gov identifier: NCT01273805 informed consent prior to aggregation,!, immunity and cancer vaccinesâ factor and decreases tissue factor outcome and risk factors Responsible Party ): Wolpin!