Extracellular DNA traps promote thrombosis. *p < 0.05 vs. Sham, **p < 0.05 vs. Tumor. PubMed  Clin Cancer Res. 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. The n for each experiment reports the number of individual animals. Neutrophil extracellular traps (NETs) occur when activated neutrophils release their intracellular contents, including DNA, histones, granules and proteins, into the surrounding tissue or circulation [12]. Neutrophil histone modification by peptidylarginine deiminase 4 is critical for deep vein thrombosis in mice. Scand J Gastroenterol. 2017;18(3):487. But then there's this: Repurposing Drugs in Oncology (ReDO)—chloroquine and hydroxychloroquine as anti-cancer agents: 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. J Thromb Haemost. NETs promote hypercoagulability in PDA by releasing circulating tissue factor. CAS  Review of correlative data from patients treated on a randomized protocol of preoperative chemotherapy with and without hydroxychloroquine demonstrated a reduction in peri-operative VTE rate from 30 to 9.1% with hydroxychloroquine that neared statistical significance (p = 0.053) despite the trial not being designed to study VTE. This study reports correlative data from two clinical trials that registered with clinicaltrials.gov, NCT01128296 (May 21, 2010) and NCT01978184 (November 7, 2013). https://doi.org/10.1186/s12885-018-4584-2, DOI: https://doi.org/10.1186/s12885-018-4584-2. Researchers in the laboratory have tested tumors from patients with pancreatic cancer and have discovered that they have certain pathways inside the cells that promote growth and survival of the tumor. These studies are designed to fail. Platelet aggregometry was performed on RAGE knockout (RAGE KO) animals, which have global genetic depletion of RAGE. Oncoimmunology. After establishing safety in a Phase I run-in, 112 patients were … Cite this article. JMO, XL, MAR, CTW, JLS, WRD, and JTE analyzed and interpreted the data and provided critical review of the manuscript. Neutrophil extracellular trap-derived enzymes oxidize high-density lipoprotein: an additional proatherogenic mechanism in systemic lupus erythematosus. NET upregulation of platelet aggregation is mediated by neutrophil DNA and platelet RAGE. Neutrophil extracellular trap (NET) impact on deep vein thrombosis. The authors declare that they have no competing interests. Plasma DNA is elevated in patients with deep vein thrombosis. Pre-tx = Pre-treatment, CCI=Charlson Comorbidity Index, EUS = Endoscopic ultrasound. Hydroxychloroquine reverses platelet activation induced by human IgG antiphospholipid antibodies. For the orthotopic pancreatic cancer model, wild type, RAGE KO and PAD4 KO mice were randomly allocated and injected with 1 × 106 Panc02 cells (National Cancer Institute repository, 2008) into the tail of the pancreas through a limited laparotomy. About the collective. Formation of ex vivo NETs. "Meaning Hydroxychloroquine added to chemotherapy did not improve overall survival among patients with metastatic pancreatic cancer." Tumor response rate is the percentage of patients achieving complete or partial response on treatment based on RECIST 1.0 criteria. PAD4-mediated neutrophil extracellular trap formation is not required for immunity against influenza infection. Kambas K, Chrysanthopoulou A, Vassilopoulos D, Apostolidou E, Skendros P, Girod A, et al. Both in vitro treatment of whole blood (Fig. 4a) and in vivo treatment of mice (Fig. 4b) with chloroquine resulted in decreased platelet aggregation and activation (Additional file 2: Figure S2C). 2014;12(12):2074–88. Neutrophils were harvested from healthy volunteer blood or murine bone marrow using density gradient centrifugation [17]. Thromboelastograms (TEGs) were performed to assess hypercoagulability and changes associated with treatment. volume 18, Article number: 678 (2018) 2015;22(13):4402–10. Annals of oncology: official journal of the European society for. Since NETs are known to release tissue factor, we evaluated levels of circulating tissue factor in our murine models of pancreatic cancer. Tissue factor, a transmembrane receptor in subendothelial cells, is a key initiator of the extrinsic coagulation cascade and is a contributor to hypercoagulability in pancreatic cancer [24]. U.S. Department of Health and Human Services, The safety and scientific validity of this study is the responsibility of the study sponsor and investigators. Furthermore, the autophagy inhibitor chloroquine inhibits NET formation [13, 14]. BMC Cancer PAD4 knockout tumor-burdened mice, unable to form NETs, had decreased aggregation and decreased circulating tissue factor. These studies were not powered to evaluate the exploratory endpoints including in the current manuscript. Samples were placed into TEG cups 2 IU of Heparinase I and 20 μL of 0.2 mol/l CaCl2 was added. Cells were initially plated in Hank’s Balanced Salt Solution (HBSS, Gibco, Grand Island, NY, USA), then to form NETs, HBSS was removed and cells were stimulated with 500 nM phorbol 12-myristate 13-acetate (PMA, Sigma, St. Louis, MO, USA) in RPMI. Study record managers: refer to the Data Element Definitions if submitting registration or results information. Additionally, RAGE knockout mice had no differences in platelet aggregation at baseline, but had decreased platelet aggregation in tumor burdened mice compared with wild type. While these findings point to extracellular DNA and RAGE promoting NET mediated platelet aggregation, there are many components released from NETs that may also have an impact on hypercoagulability and were not evaluated in the current analysis. During NET formation, PAD4 mediated histone citrullination leads to unwinding and release of DNA from neutrophils [37]. Google ScholarÂ. Arterioscler Thromb Vasc Biol. Correspondence to Slides were washed three times with BSA solution and incubated for 1 h at RT with Alexa 488 donkey anti mouse secondary antibody (A21202, Invitrogen) diluted 1:500, combined with donkey anti rabbit CY3 (711–165-152, Jackson Immuno) 1:1000, and donkey anti sheep Cy5 (713–175-147, Jackson) in BSA solution. Hydroxychloroquine may inactivate these pathways and results in the death of pancreatic cancer cells. 2014;34(9):1977–84. ], Grade 4-5 Treatment-Related Toxicity [ Time Frame: Adverse events were assessed each cycle throughout treatment. 1974;3(5923):94–5. Mice from tumor bearing animals demonstrated significantly greater platelet aggregation in response to collagen stimulation (Fig. 1a) and had heightened platelet activation as measured by %CD62P positive platelets (Additional file 1: Figure S1A). Data is reported as the area under the curve (AUC), which incorporates both the slope and amplitude of the aggregation curve. Additionally, the 90 day postoperative reduction in VTE occurred despite HCQ stopping at time of surgery. Priming of neutrophils toward NETosis promotes tumor growth. The human F3/CD142/Tissue factor ELISA kit was used to measure tissue factor in patient blood samples (LS Bio, LS-F433). J Exp Med. Google ScholarÂ. Arterioscler Thromb Vasc Biol. Google ScholarÂ. Brinkmann V, Reichard U, Goosmann C, Fauler B, Uhlemann Y, Weiss DS, et al. Toll-like receptor 4 regulates platelet function and contributes to coagulation abnormality and organ injury in hemorrhagic shock and resuscitation. Springer Nature. 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). To determine the role of NETs in platelet aggregation in our cancer model, we first examined platelet activation and aggregation in mice injected with orthotopic tumor and sham injected controls. Progression-free survival based on the Kaplan-Meier method is defined as the duration of time from study entry to time of objective progression on CT scan or the time of death for patients with clinical deterioration resulting in withdrawal from the trial. Int J Mol Sci. Sirois CM, Jin T, Miller AL, Bertheloot D, Nakamura H, Horvath GL, et al. Fuchs TA, Brill A, Duerschmied D, Schatzberg D, Monestier M, Myers DD Jr, et al. Circ Cardiovasc Genet. Rafael Pharmaceuticals, a company specializing in the field of cancer metabolism, has obtained orphan drug stats from the US Food and Drug Administration (FDA) for CPI-613 (devimistat). Paricalcitol (a form of vitamin D) works by blocking a signal in the cancer cells that leads to growth and spreading of the tumor. BMC Cancer 18, 678 (2018). Ann Surg Oncol. PD for the evaluation of non-target lesions is the appearance of one or more new lesions and/or unequivocal progression of non-target lesions. Chloroquine and hydroxychloroquine decrease CXCL12-mediated proliferation in pancreatic cancer cell lines. PubMed Central  CAS  Ann Rheum Dis. Levels of circulating tissue factor, the initiator of extrinsic coagulation, were measured using ELISA. 2007;13(10):2870–5. PubMed  Kambas K, Mitroulis I, Apostolidou E, Girod A, Chrysanthopoulou A, Pneumatikos I, et al. Cookies policy. A more recent trial randomized patients to two cycles of preoperative gemcitabine/nab-paclitaxel with or without 1200 mg/day oral hydroxychloroquine (UPCI 13–074, IRB Protocol #13080444). Please refer to this study by its ClinicalTrials.gov identifier (NCT number): NCT01273805. Please remove one or more studies before adding more. In our recent randomized trial evaluating two months of preoperative hydroxychloroquine treatment in patients with pancreatic cancer, the VTE rate was lower in patients receiving HCQ compared to patients receiving gemcitabine/nab-paclitaxel alone. We evaluated hydoxychloroquine (HCQ), an inhibitor of autophagy, in patients with pancreatic cancer and analyzed pharmacodynamic markers in treated patients and mice. Per RECIST 1.0 criteria: progressive disease (PD) is at least a 20% increase in the sum of longest diameter (LD) of target lesions taking as reference the smallest sum LD recorded since the treatment started or the appearance of one or more new lesions. 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. 2013;122(11):1873–80. Kruger S, Haas M, Burkl C, Goehring P, Kleespies A, Roeder F, et al. 2016;76(6):1367–80. 1975;41(12):761–6. Oncologist. ERK inhibitors stop the signal that a cancer cell receives telling it to grow. 2017;24(12):1600-6. Trametinib may stop the growth of tumor cells by blocking some of the enzymes needed for cell growth. Department of Surgery, University of Pittsburgh, Pittsburgh, PA, USA, Brian A. Boone, Pranav Murthy, Jennifer Miller-Ocuin, W. Reed Doerfler, Jarrod T. Ellis, Xiaoyan Liang, Jason L. Sperry, Michael T. Lotze, Matthew D. Neal & Herbert J. Zeh III, Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, PA, USA, Departments of Thoracic Surgery, University of Pittsburgh, Pittsburgh, PA, USA, Immunology, University of Pittsburgh, Pittsburgh, PA, USA, Bioengineering, University of Pittsburgh, Pittsburgh, PA, USA, UPMC Cancer Pavilion, University of Pittsburgh, Suite 417, 5150 Centre Ave, Pittsburgh, PA, 15232, USA, You can also search for this author in Che-Motherapy ( 11, 12 ) of recurrent venous thromboembolism in malignancy: biomarker analyses the. Of study participation for the treatment of cancer patients treated with preoperative gemcitabine/nab-paclitaxel with and hydroxychloroquine. Jancinova V, et al, 5 ] is reported as the area under the curve ( AUC ) which! Js, Mathew a, Mezouar S, Lewis C, Fauler B, PS... Swystun LL, Dwivedi DJ, et al hypercoagulability was the coagulation (... Sham, Triangle = tumor autophagy, for pancreatic cancer and thromboembolic disease a... 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