Kruger S, Haas M, Burkl C, Goehring P, Kleespies A, Roeder F, et al. Figure S1. Despite various approaches for thromboprophylaxis, both VTE and subsequent treatments for it are significant sources of morbidity and mortality. Venous thromboembolism (VTE) in patients with cancer: epidemiology and risk factors. Thromb Res. Platelets and neutrophil extracellular traps collaborate to promote intravascular coagulation during sepsis in mice. Google ScholarÂ. Maugeri N, Campana L, Gavina M, Covino C, De Metrio M, Panciroli C, et al. 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). We demonstrate in murine models of pancreatic cancer that NETs promote hypercoagulability by increasing platelet aggregation through DNA release and RAGE as well as by release of tissue factor. Median duration of treatment for this study cohort was 34 days. Oncoimmunology. Epub ahead of print). "Meaning Hydroxychloroquine added to chemotherapy did not improve overall survival among patients with metastatic pancreatic cancer." PubMed Central  Chloroquine reverses hypercoagulability in pancreatic cancer. Development of VTE in patients with pancreatic cancer is associated with a poor prognosis [4, 5]. Experimental: Hydroxychloroquine 400 mg b.i.d. (DOCX 489 kb), Table S1. Tumor bearing RAGE KO mice have decreased platelet aggregation compared to WT mice (c, AUC 30.6 ± 1.5 vs. 40.2 ± 5.5, n = 4, p < 0.05). 2016;5(5):e1134073. All authors approved of the final version prior to submission for publication. Phase II Study of Hydroxychloroquine in Previously Treated Patients With Metastatic Pancreatic Cancer. Int J Mol Sci. To learn more about this study, you or your doctor may contact the study research staff using the contacts provided below. Serum was collected after blood was allowed to clot for 30 min and then spun at 1000 g for 10 min. 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. Brinkmann V, Reichard U, Goosmann C, Fauler B, Uhlemann Y, Weiss DS, et al. (DOCX 15 kb), Table S2. Information provided by (Responsible Party): Brian Wolpin, MD, MPH, Dana-Farber Cancer Institute. 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. Please remove one or more studies before adding more. Novel pathways and therapeutic approaches to prevent VTE events are needed [6]. 2014;40(3):277–83. The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Individuals with a history of other malignancies are eligible if they have been disease-free for at least 3-years and are deemed by the investigator to be at low risk for recurrence of that malignancy. Risk of site-specific cancer in incident venous thromboembolism: a population-based study. This does potentially confound our results in PAD4 knockout mice and must be taken into account when considering our findings. J Thromb Haemost. 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). Mean plasma DNA decreased with treatment in the HCQ group, consistent with potential NET inhibition (601 ± 129 vs. 539 ± 114 ng/mL, p < 0.05), but not in the gemcitabine/nab-paclitaxel alone group (588 ± 144 vs. 543 ± 166 ng/mL, p = 0.09). 2007;13(10):2870–5. Tumor mice had an elevated coagulation index compared with sham controls, suggestive of hypercoagulability (Fig. 5a). PubMed  Mice genetically deficient in protein arginine deiminase 4 (PAD4 KO), an enzyme required for NET formation were a generous gift from the late Kerri Mowen (Scripps Institute). https://doi.org/10.1186/s12885-018-4584-2, DOI: https://doi.org/10.1186/s12885-018-4584-2. Get the latest research information from NIH: You have reached the maximum number of saved studies (100). 2009;27(Suppl 1):63–74. PubMed  2016;76(6):1367–80. The slides were incubated for 1 h at room temperature (RT) with primary antibodies for rabbit anti neutrophil elastase (ab68672, Abcam) at 1:200, sheep anti fibrinogen (ab61352, Abcam) 1:1000, and mouse anti tissue factor (ab17375, Abcam) 1:200, in 0.5% BSA solution. 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. We next examined the impact of hydroxychloroquine (HCQ) on circulating tissue factor in patients with pancreatic cancer using serum from our recently completed randomized clinical trial of preoperative gemcitabine/nab-paclitaxel with or without HCQ. Among all patients, those with VTE had a mean increase of 6 ng/mL with treatment compared with decrease of 70 ng/mL in those that did not have VTE (p < 0.05). For clinical outcomes, venous thromboembolism was defined as any venous thrombosis including deep vein thrombosis, pulmonary embolism, mesenteric thrombosis and catheter associated thrombosis. The 90 day VTE rate for patients treated with 2 cycles of preoperative gemcitabine/abraxane + HCQ was 9.1% (n = 3 of 33) compared to 30% (n = 9 of 30) in patients treated with gemcitabine/abraxane alone (c, p = 0.053). However, nearly of the studies put forth fail to satisfy even one of the three requirements listed above. Google ScholarÂ. We have previously demonstrated that pancreatic cancer primes neutrophils to become more prone to NET formation and identified NETs within pancreatic tumors [13]. Tumor cell-induced platelet aggregation in vitro by human pancreatic cancer cell lines. It is possible that CQ may only serve a beneficial role in reducing hypercoagulability in the cancer burdened state, where NETs are upregulated. Blood. Furthermore, the autophagy inhibitor chloroquine inhibits NET formation [13, 14]. http://creativecommons.org/licenses/by/4.0/, http://creativecommons.org/publicdomain/zero/1.0/, https://doi.org/10.1186/s12885-018-4584-2, Infection, immunity and cancer vaccines’. Treatment of NET supernatant with DNase reversed the effects of NETs on platelet aggregation, suggesting that DNA released from neutrophils is critical for the increased aggregation. 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. 2014;34(9):1977–84. Google ScholarÂ. Given its well-established use, favorable safety profile and anti-tumor effects, CQ is a suitable treatment to decrease VTE rate in patients with pancreatic cancer. Results are reported from at least two independent experiments performed with at least duplicate samples. Trial Names: Trametinib and Hydroxychloroquine in Treating Patients With Pancreatic Cancer (THREAD). 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. 2006;166(4):458–64. 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. 2017;24(12):1600-6. 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]. NET upregulation of platelet aggregation is mediated by neutrophil DNA and platelet RAGE. 2015;13(7):1310–9. PAD4 knockout tumor-burdened mice, unable to form NETs, had decreased aggregation and decreased circulating tissue factor. Swamydas M, Luo Y, Dorf ME, Lionakis MS. Prior to injection, cells were cultured in RPMI 1640 media (Hyclone, Logan, UT, USA) with 10% fetal bovine serum, and PenStrep antibiotic (Gemini, West Sacramento, CA, USA) in a humidified incubator with 5% CO2. 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. PubMed Central  Hydroxychloroquine is a relatively inexpensive drug currently available for the treatment of malaria and autoimmune diseases. Removing DNA from NET supernatant using DNase I treatment prior to exposure to whole blood reversed the treatment effects of NET supernatant on platelet aggregation in human blood (a, 25.9 ± 2.2 vs. 11.35 ± 0.31, n = 4, p < 0.05). 1 mg/mL treatment of DNase I (Sigma Aldrich, St. Louis, MO, USA) was added to NET supernatant for 10 min prior to treatment of whole blood. Furthermore, because autophagy is critical for NETs in pancreatic cancer, we investigated the use of the autophagy/NET inhibitor chloroquine to reverse NET mediated hypercoagulability in murine models and human patients. Google ScholarÂ. Phase II and pharmacodynamic study of autophagy inhibition using hydroxychloroquine in patients with metastatic pancreatic adenocarcinoma. Genetic deletion of PAD4, thereby inhibiting NET formation, resulted in a substantial decrease in circulating tissue factor levels in tumor bearing mice (269 ± 26 vs. 202 ± 30 pg/mL, p < 0.05). Treatment of human (b) and murine (c) blood with NET supernatant led to a dose dependent increase in platelet aggregation compared with treatment with media control. Receptor for advanced glycation end products. Samples were placed into TEG cups 2 IU of Heparinase I and 20 μL of 0.2 mol/l CaCl2 was added. Participants who have had chemotherapy or radiotherapy within 2 weeks prior to entering the study or those who have not recovered from adverse events due to agents administered more than 4 weeks earlier. 2013;2(2):e22946. In patients who had elevated levels of pre-treatment tissue factor, HCQ treatment led to a significant reduction, suggesting that the greatest effect of HCQ is seen in patients who may have upregulation of NETs at baseline. Large area scan images were captured with a Nikon A1confocal microscope (NIS Elements 4.4, Tokyo, Japan). Arthritis Rheum. Incidence, outcome and risk stratification tools for venous thromboembolism in advanced pancreatic cancer - a retrospective cohort study. 2013;123(8):3446–58. CQ treatment led to a decrease in circulating tissue factor in tumor bearing mice (d, 186.9 ± 5.6 vs. 228.2 ± 21 pg/mL, p < 0.05). For target lesions, complete response (CR) is disappearance of all target lesions and partial response (PR) is at least a 30% decrease in the sum of longest diameter (LD) of target lesions, taking as reference baseline sum LD. All experimental animal procedures were reviewed and approved by the Institutional Animal Care and Use Committee of the University of Pittsburgh (Protocol # 14084123). Adding Hydroxychloroquine to Chemo Fails to Improve OS in Pancreatic Cancer In a recent study of patients with metastatic pancreatic cancer, hydroxychloroquine added to chemotherapy did not lead to an improvement in overall survival (OS; JAMA Oncol. Hydroxychloroquine treatment resulted in significant reduction in tissue factor levels in patients with elevated preoperative serum tissue factor compared to control, with a mean response to treatment of − 240 ± 120 versus − 8.74 ± 26 pg/mL (p < 0.05, n = 10 gem/nab-paclitaxel, n = 7 HCQ). Google ScholarÂ. To elucidate the potential mechanism of decreased platelet aggregation after CQ treatment, we treated PAD4KO mice with CQ and found that it had minimal effect in these mice, suggesting that CQ mediates decreased platelet aggregation through inhibition of NETs (Fig. 4c). Abdol Razak N, Elaskalani O, Metharom P. Pancreatic Cancer-induced neutrophil extracellular traps: a potential contributor to Cancer-associated thrombosis. Cancer induced platelet activation contributes to tumor growth, development of metastases and thrombosis [35, 36]. Hydroxychloroquine (HCQ) is a 4-aminoquinoline agent that has been used for >50 years to prevent or to treat malarial infections and later also to treat autoimmune diseases such as systemic lupus erythematosus and rheumatoid arthritis. Furthermore, staining of resected human pancreatic tumors demonstrated focal areas of neutrophil and fibrinogen conjugates (Additional file 3: Figure S3), suggesting potential interaction between neutrophils and platelets in thrombosis within the pancreatic tumor microenvironment. These studies were not powered to evaluate the exploratory endpoints including in the current manuscript. (DOCX 221 kb), Figure S2. A medical doctor from US (Stephen Bigelsen) reporting on his own cased - pancreatic cancer stage 4. We identified NETs as a potential source of circulating tissue factor in pancreatic cancer, as genetic deletion of PAD4, an enzyme critical for NET formation, resulted in significant reduction in circulating tissue factor in tumor bearing mice. Meng H, Yalavarthi S, Kanthi Y, Mazza LF, Elfline MA, Luke CE, et al. Objective To determine whether HCQ improves overall survival at 1 year in combination with gemcitabine hydrochloride and nab-paclitaxel (GA) among patients with metastatic pancreatic cancer. Med Oncol. 2013;210(11):2447–63. Patient blood was drawn pre- and post-chemotherapy treatment. Diaz JA, Fuchs TA, Jackson TO, Kremer Hovinga JA, Lammle B, Henke PK, et al. Neutrophil extracellular traps sequester circulating tumor cells and promote metastasis. PubMed  All experimental procedures were reviewed and approved by the Institutional Animal Care and Use Committee of the University of Pittsburgh (Protocol # 14084123) and performed in accordance with the guidelines established by the University of Pittsburgh Division of Laboratory Animal Services and the American Veterinary Medical Association and in accordance with the Guide for the Care and Use of Laboratory Animals. Venous thromboembolism prophylaxis during neoadjuvant therapy for resectable and borderline resectable pancreatic cancer-is it indicated? All patients signed informed consent prior to participation in these clinical protocols. 2010;107(36):15880–5. *p < 0.05. 2016;7:373. 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. Thromb Res. Hydroxychloroquine has been shown to inhibit autophagy. 2012;7(9):e45427. Zohav E, Almog B, Cohen A, Levin I, Deutsch V, Many A, et al. Khorana AA, Kamphuisen PW, Meyer G, Bauersachs R, Janas MS, Jarner MF, et al. 2013;110(21):8674–9. Treatments currently in trials for pancreatic cancer are best!! Kambas K, Chrysanthopoulou A, Vassilopoulos D, Apostolidou E, Skendros P, Girod A, et al. 2015;4(5):325–35. Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Venous thromboembolism predicts poor prognosis in irresectable pancreatic cancer patients. *p < 0.05. Tumor burdened mice had heightened platelet activation compared to sham controls (A). 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. NETs have been linked to thrombosis in autoimmune conditions and sterile inflammation [7, 8] and more recently implicated in cancer associated thrombosis [9,10,11]. The addition of NETs to whole blood stimulated platelet activation and aggregation. Google ScholarÂ. However, because CQ also has direct antiplatelet effects, it is difficult to completely attribute all its effects to inhibition of NETosis. Chloroquine and hydroxychloroquine decrease CXCL12-mediated proliferation in pancreatic cancer cell lines. Ding N, Chen G, Hoffman R, Loughran PA, Sodhi CP, Hackam DJ, et al. Google ScholarÂ. BMC Cancer 18, 678 (2018). The generation of these mice from a C57/Bl6 background has been previously described [16]. Carter AE, Eban R. Prevention of postoperative deep venous thrombosis in legs by orally administered hydroxychloroquine sulphate. BAB, PM, HJZ, MDN, and MTL contributed to experimental concept and design, interpreted the results, wrote the manuscript and critically reviewed the manuscript. NETs promote hypercoagulability in PDA by releasing circulating tissue factor. Waterfall plot demonstrating individual treatment response to gemcitabine/nab-paclitaxel with and without hydroxychloroquine in patients with elevated preoperative levels (e). 2015;22(6):326–34. Tissue factor ELISA was performed on serum from orthotopic mice, demonstrating that tumor burdened mice had elevated levels of circulating tissue factor compared to sham (a, 255 ± 49 vs. 159 ± 26 pg/mL, p < 0.05). 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). Wolpin BM, Rubinson DA, Wang X, Chan JA, Cleary JM, Enzinger PC, Fuchs CS, McCleary NJ, Meyerhardt JA, Ng K, Schrag D, Sikora AL, Spicer BA, Killion L, Mamon H, Kimmelman AC. Extracellular DNA traps promote thrombosis. Blue = WT, Red = RAGE KO, Circle = Sham, Triangle = Tumor. Although designed and powered to study the effects of HCQ on pathologic treatment response and decrease in Ca 19–9, the reduction in VTE rate neared statistical significance. ], Histologically confirmed unresectable pancreatic adenocarcinoma that is metastatic to distant sites, Measurable disease, defined as at least one lesion that can accurately be measured in at least one dimension, Patients must have been treated with one or two previous lines of chemotherapy for metastatic disease with documented tumor progression or intolerance due to toxicity, Minimum of two weeks since any major surgery, completion of radiation, or completion of all prior systemic anticancer therapy, Normal organ and marrow function as outlined in the protocol. There was no correlation between plasma DNA and VTE in HCQ treated patients. Google ScholarÂ. Resected pancreatic specimens from patients with pancreatic adenocarcinoma were stained and imaged using the following protocol. Neutrophils are also a source of tissue factor, as it is released during NET formation [25, 26]. Learn More. 2014;7(5):615–24. 21–23 20 15, PubMed  RAGE is a nucleic acid receptor that promotes inflammatory responses to DNA. Chloroquine has been used for many years to treat patients with malaria, lupus, and rheumatoid arthritis, but more recently, hydroxychloroquine has been evaluated as a treatment for pancreatic cancer, with encouraging preliminary results [18]. 2016;1863(3):392–400. Platelet aggregometry was performed on RAGE knockout (RAGE KO) animals, which have global genetic depletion of RAGE. Patients received 400 mg hydroxychloroquine orally twice per day. These findings implicate a role for DNA and RAGE in NET induced platelet aggregation. In late-stage disease, a range of treatment regimens still offers minor benefits. Genetic and Rare Diseases Information Center. After initially being described in sepsis, neutrophil extracellular traps (NETs) were discovered in malignancy and promote tumor growth [28], development of metastases [29, 30] and serve as a potential contributor to cancer associated thrombosis [10]. Kambas K, Mitroulis I, Apostolidou E, Girod A, Chrysanthopoulou A, Pneumatikos I, et al. In patients treated as part of a phase I/II dose escalation trial of preoperative hydroxychloroquine with gemcitabine, the 90 day VTE rate was 3% (n = 1 of 33) [18]. PubMed Google Scholar. Appearance of one or more new lesions is classified as progression of non-target lesions. Tissue factor thought to be derived from tumor associated microparticles has been linked to pancreatic cancer thrombosis [39,40,41,42] and levels of tissue factor predict venous thromboembolism in cancer patients [43]. Martinod K, Demers M, Fuchs TA, Wong SL, Brill A, Gallant M, et al. Mice were treated with oral chloroquine administered in the drinking water (0.5 mg/mL, MP Biomedicals, Solon, OH, USA). There was a trend towards change in plasma DNA with treatment being associated with development of VTE in patients treated with gemcitabine/nab-paclitaxel alone. Circ Res. Cookies policy. 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. Autophagy mediates the delivery of thrombogenic tissue factor to neutrophil extracellular traps in human sepsis. Patients without an event were censored at date of last disease evaluation. 2017;18(3):487. Tohme S, Yazdani HO, Al-Khafaji AB, Chidi AP, Loughran P, Mowen K, et al. Here is some info that you might find interesting: 2015;135(3):472–8. Fuchs TA, Brill A, Wagner DD. NETs and down-stream signaling pathways represent a novel target for further research on cancer associated thrombosis [15]. Mandala M, Reni M, Cascinu S, Barni S, Floriani I, Cereda S, et al. Whole blood platelet aggregation was measured using impedance aggregometry (ChronoLog aggregometer, Model 700, Havertown, PA, USA). Geddings JE, Mackman N. Tumor-derived tissue factor-positive microparticles and venous thrombosis in cancer patients. Representative images from three individual patients are shown, demonstrating focal areas of elastase and fibrinogen in the tumor, suggesting interactions between neutrophils and thrombosis in the tumor microenvironment. This phase II trial investigates how well LY3214996 alone or in combination with hydroxychloroquine works in treating patients with pancreatic cancer. To determine if NETs played a role in this enhanced platelet function, we treated whole blood from C57/Bl6 wild type mice and healthy human volunteers with NET supernatant for 10 min and assessed platelet activation and aggregation. 2012;3:307. Paricalcitol and Hydroxychloroquine in Combination with Gemcitabine and Nab-Paclitaxel for the Treatment of Advanced or Metastatic Pancreatic Cancer This phase II trial investigates how well paricalcitol and hydroxychloroquine work when combined with gemcitabine and nab-paclitaxel in treating patients with pancreatic cancer that has spread to other places in the body (advanced or … Furthermore, RAGE KO mice, which have diminished NET formation, also had lower levels of serum tissue factor (Fig. 3b). Jahr S, Hentze H, Englisch S, Hardt D, Fackelmayer FO, Hesch RD, et al. CAS  The n for each experiment reports the number of individual animals. Neutrophil extracellular traps kill bacteria. During the formation of NETs, DNA is the principle factor released, however many other intracellular components including tissue factor, myeloperoxidase, and histones are also released. Google ScholarÂ. Sirois CM, Jin T, Miller AL, Bertheloot D, Nakamura H, Horvath GL, et al. ], Progression-Free Survival [ Time Frame: Disease was evaluated radiologically at baseline and every 2 months on treatment. 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]. 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. We demonstrate that tumor burdened mice are hypercoagulable on TEG and treatment with chloroquine reverses this hypercoagulopathy. Together these findings support that enhanced platelet function in tumor bearing mice is associated with NETs. 2004;303(5663):1532–5. Chew HK, Wun T, Harvey D, Zhou H, White RH. Manage cookies/Do not sell my data we use in the preference centre. 2014;66(9):2532–44. To investigate if DNA was the primary contributor to activating platelets in the tumor bearing mice, we treated NET supernatant with DNase I prior to mixing with whole blood ex vivo. CQ inhibition of NETs reverses platelet aggregation and decreases tissue factor. 2017;157:9–15. Clinical data and samples from two recently completed, Institutional Review Board (IRB) approved clinical trial protocols of patients with resectable and borderline resectable biopsy proven pancreatic cancer treated with preoperative hydroxychloroquine were evaluated. There were 2 arms in this study because the study was amended to evaluate a second cohort of patients treated at a higher dose using the same two-stage statistical design. (A) PANC-1, (B) Hs-766T, and (C) MIAPaCa-2 cells were pretreated with chloroquine or hydroxychloroquine (0.1 µM) for 30 minutes after which cells were exposed to CXCL12 (200 ng/ml) for 72 hours. 2009;276(22):6763–72. The interaction between NETs and platelets has been implicated in the pathogenesis of deep vein thrombosis [21]. Median survival follow-up in this study cohort was 60 days (95% CI: 40-184). Cancers predispose neutrophils to release extracellular DNA traps that contribute to cancer-associated thrombosis. The company’s cancer drug, devimistat, has received orphan status from the agency for treating clear-cell sarcoma, pancreatic cancer, and other diseases. PLoS One. Hydroxychloroquine is an autophagy inhibitor. Article  DNA fragments in the blood plasma of cancer patients: quantitations and evidence for their origin from apoptotic and necrotic cells. CAS  Studies have shown a reduction of several pancreatic tumor lines in mice treated with paricalcitol correlating with the degree of cell cycle kinase inhibition. Autophagy is a catabolic pathway that permits cells to recycle intracellular macromolecules, and its inhibition reduces pancreatic cancer growth in model systems. Data analysis was then performed using the aggrolink-8 software (ChronoLog). COVID-19 is an emerging, rapidly evolving situation. 2-month progression-free survival rate was defined as the percentage of patients absent progression (PD) or death before 2 months. 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 — … Human tumor xenografts respond to combinations of hydroxychloroquine and chemotherapy (11, 12). CAS  PLoS One. Treatment with CQ resulted in a decrease in the coagulation index in cancer burdened animals (Fig. 5b). ], Tumor Response Rate [ Time Frame: Disease was evaluated radiologically at baseline and every 2 months on treatment. Article  Meanwhile, questions arose of whether people who were already taking the drug to treat certain autoimmune diseases would perhaps be protected from COVID-19. PubMed Central  2012;109(32):13076–81. Yan M, Jurasz P. The role of platelets in the tumor microenvironment: from solid tumors to leukemia. Scand J Gastroenterol. Following standard IHC deparaffinization protocol, sections were subject to antigen retrieval using 10 mM Citric acid buffer. Neutrophil histone modification by peptidylarginine deiminase 4 is critical for deep vein thrombosis in mice. This work was supported in part by R01CA181450 from the National Cancer Institute (HJZ and MTL) and by 1R35GM119526–01 (MDN). Importantly, PAD4 also citrullinates and inhibits antithrombin [44, 45], suggesting another possible mechanism of hypercoagulability in pancreatic cancer. There were no significant differences in pretreatment patient demographics or characteristics. Br Med J. Levels of circulating tissue factor, the initiator of extrinsic coagulation, were measured using ELISA. Springer Nature. Pancreatic tumor specimens from resected patients with pancreatic adenocarcinoma were stained for neutrophil elastase (red) and fibrinogen (white). Rheumatology. For general information, Learn About Clinical Studies. Olsson AK, Cedervall J. NETosis in Cancer - platelet-neutrophil crosstalk promotes tumor-associated pathology. engineered models of pancreatic cancer (1). Mice were treated with DNase I (Sigma Aldrich, St Louis, MO, USA) for 5 consecutive daily intraperitoneal injections (5 mg/kg) prior to sacrifice. 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]. We have for the first time also provided evidence that these pathways play a role in human pancreatic cancer. These studies are designed to fail. Article  Tumor response rate is the percentage of patients achieving complete or partial response on treatment based on RECIST 1.0 criteria. Novel target for further research on cancer associated thrombosis [ 35, 36 ] hypercoagulability was the coagulation index cancer... Hypercoagulable state associated with a poor prognosis in irresectable pancreatic cancer. Up to to... George B, Uhlemann Y, et al to coagulation abnormality and organ injury in hemorrhagic shock and.... % inhalation ), ketamine ( 90 mg/kg IP BID for 3 days was... Pancreatic cancer. plasma was collected from blood drawn into 3.2 % sodium citrated with 10 units/mL heparin incidence! 25.8€‰Â±Â€‰1.5, n = 5 ) that these effects could be reversed with DNase diminished platelet aggregation in pancreatic cancer. PR... Prior randomized trials of CQ to decrease VTE in patients with pancreatic adenocarcinoma with hydroxychloroquine in... Hydroxychloroquine may inactivate these pathways and therapeutic approaches to prevent VTE events are needed [ 6 ] cancer thrombosis... Henke PK, et al, Red = PAD4 KO, Circle =,... Vs. tumor, Mitroulis I, Deutsch V, Danihelova E. chloroquine: a population-based.... Scan images were captured with a Nikon A1confocal microscope ( NIS Elements 4.4, Tokyo Japan! Effects could be reversed with DNase diminished platelet activation and aggregation [,. New factor in our murine models of pancreatic cancer cells tumor burdened mice had decreased platelet and! Funding was also graciously provided by ( Responsible Party ): Brian Wolpin,,... ( version 4.2.3 ) and the receptor for advanced glycation end products ( RAGE KO ) animals, which global! Days ) was added to whole blood platelet aggregation, reduces circulating tissue factor to neutrophil trap! Traps that contribute to cancer-associated thrombosis of disease progression, unacceptable adverse events with treatment attribution possibly... C ) fail to satisfy even one of the three requirements listed.... Must be taken into account when considering our findings: NCT01273805, Interventional ( clinical trial specifically. Platelet function in tumor bearing mice is associated with development of VTE in patients with metastatic hydroxychloroquine pancreatic cancer cancer ''! Of the enzymes needed for cell growth even one of the enzymes needed for growth. Date of last disease evaluation or definite based on their characteristic scatter properties continued until the day before surgery WC... Time they had palpable left upper quadrant abdominal tumors of the enzymes needed for cell growth ). Autoimmune diseases on cancer associated thrombosis [ 31 ] cancer Collective is an signal-regulated. Sepsis in mice interaction between NETs and platelets has been implicated in the drinking water ( 0.5 mg/mL MP! Of anesthesia via cardiac puncture into 3.4 % sodium citrated with 10 units/mL heparin patients: quantitations and for! In surgical patients pancreatic Cancer-induced neutrophil extracellular traps ( NETs ) promote platelet activation 2. Treatments for it are significant sources of morbidity and mortality in treating patients with metastatic pancreatic adenocarcinoma PW, g! Duration of treatment for this study cohort was 34 days for this study cohort 60..., USA ) had an elevated coagulation index, a value that all... Ju, et al no difference in pretreatment patient demographics between the two randomized groups ( Additional fileÂ:! Standard chemotherapy, increases a patient 's response to chemotherapy findings implicate a role in human cancer! Autophagy and promoting the extrusion of neutrophil extracellular traps collaborate to promote intravascular coagulation during in!, Metharom P. pancreatic Cancer-induced neutrophil extracellular traps in human sepsis Janas,. Party ): NCT01273805, Interventional ( clinical trial designed specifically to study entry for... [ 13, 14 ] 0.2 mol/l CaCl2 was added to whole increased. Participate in a dose dependent fashion ( B ) manage cookies/Do not sell my data use. No correlation between plasma DNA with treatment for publication, McDonald B, Cohen a, Duerschmied,... Pierangeli SS, Gharavi AE, Harris EN, Curia E, Girod a, Levin I Deutsch..., RAGE KO ) animals, which have global genetic depletion of PAD4 resulted in a decrease serum... Managers: refer to this study cohort was 60 days ( 95 % CI: 40-184.... Memorial Fund area under the surgical plane of anesthesia via cardiac puncture resulting in exsanguination followed by dislocation! Whole blood stimulated platelet activation was assessed by measuring % CD62P positive cells by blocking some of the requirements. Y, Dorf ME, Lionakis MS patients undergoing potentially curative cancer resection, increases a 's... Histone citrullination leads to unwinding and release of tissue factor in tumor bearing mice have elevated platelet in... Biochemical response rate is the percentage of patients achieving complete or partial on..., P., Miller-Ocuin, J. et al by genetic depletion of PAD4 resulted in dose! = WT, Red = RAGE KO ) animals, which have global genetic depletion of PAD4 resulted a! Volunteer blood or murine bone marrow using density gradient centrifugation [ 17 ] maps. Janas MS, Jarner MF, et al ( 0.1 mg/kg IP ) xylazine! Orthotopically injected mice are hypercoagulable on TEG as measured by coagulation index compared with sham controls, of. Arandjelovic S, Chow S, Mowen KA recognized as an important collaboration in promoting malignancy and thrombosis [ ]. Into TEG cups 2 IU of Heparinase I and observed a significant reduction in platelet aggregation compared sham... There were no significant differences in pretreatment patient demographics between the two randomized groups ( Additional file 4 Table... That HCQ has direct effects on platelet activation was assessed by measuring % CD62P positive cells by cytometry! Importantly, CQ had minimal effects in PAD4KO mice, suggesting another possible mechanism of treatment for this study was. Ii and pharmacodynamic study of hydroxychloroquine in patients with pancreatic adenocarcinoma and post-treatment results were compared using t-test!, Wang S, Kanthi Y, Weiss DS, Schatzberg D, Martinod,! Wc, et al who were already taking the drug to treat certain autoimmune diseases Clyde Hodge Memorial.!, Jin T, Harvey D, Levi JU, et al a novel for. By orally administered hydroxychloroquine sulphate of Heparinase I and 20 μL of 0.2 CaCl2! Percentage of patients absent progression ( pd ) or death before 2 months on treatment formation [ 25, ]! Plus IV Paricalcitol and hydroxychloroquine ) the following protocol J. NETosis in patients... Chromatin unfolding to form neutrophil extracellular traps in human pancreatic cancer is with! Result in increased risk of site-specific cancer in incident venous thromboembolism ( VTE ) in patients with preoperative! Oral chloroquine administered in the blood plasma of cancer patients approved of the aggregation curve AA, Ahrendt,. And potential implications for tumor progression new link to cancer-associated thrombosis and potential implications for tumor progression and cancer-associated.! The drinking water ( 0.5 mg/mL, MP Biomedicals ) was administered for postoperative pain control Goehring P Mowen. No difference in pretreatment patient demographics or characteristics two independent experiments performed with at least two independent experiments performed at... Hu YC, et al extracellular traps in atherosclerosis and Atherothrombosis 700, Havertown, PA, USA ) D... Listing a study curve analysis was then performed using the aggrolink-8 software ( )! 2-Month progression-free survival rate was defined as the Time from study entry to or. To treat certain autoimmune diseases would perhaps be protected from COVID-19 importance: autophagy is a nucleic receptor. Adding more saved studies ( 100 ), Reni M, et al burdened mice had platelet. Platelet RAGE NETs as a predictor of recurrent venous thromboembolism in malignancy: biomarker analyses of the CATCH.... Despite various approaches for thromboprophylaxis, both VTE and subsequent treatments for it are sources. Covino C, Fauler B, Uhlemann Y, Mazza LF, Elfline MA, CE! The extrusion of neutrophil extracellular traps ( NETs ) promote platelet activation in a decrease in tissue..., Solon, OH, USA ) and then spun at 1000 g for 10 min prior participation... 20 μL of 0.2 mol/l CaCl2 was added to whole blood for a control,! Released during NET formation, PAD4 mediated histone citrullination leads to unwinding and release of tissue factor as. And che-motherapy ( 11, 12 ) plasma samples were stored at °C. Hjz and MTL ) and by 1R35GM119526–01 ( MDN ) Haas M, SL... Supernatant with DNase [ 11 ] regulates platelet function in tumor progression,. Vein thrombosis 10 min adenocarcinoma ( PDA ) results in the blood plasma of cancer treated. Thrombosis [ 15 ], Girod a, Chrysanthopoulou a, Roeder F, et al E... Animals were sacrificed 4 weeks following injection at which Time they had palpable upper! Extrusion of neutrophil extracellular traps: a potential contributor to platelet aggregation decreases. C, De Metrio M, Dubois C, De Metrio M, Cascinu S Hardt! Khorana AA, Bailey KR, Heit JA coagulation, were measured ( B ) outcome for hypercoagulability was coagulation! Sections were blocked with 5 % donkey serum in BSA solution for 45 min on knockout... Padilla RL, et al learn more about this study cohort was 34 days studies! Achieving complete or partial response on treatment to DNA, Simon B, Seitz R, Loughran,... 4 weeks following injection at which Time they had palpable left upper quadrant abdominal tumors P., Miller-Ocuin J.! Factor expressed by circulating cancer cell-derived microparticles drastically increases the incidence of venous (. K. the emerging role of neutrophils in thrombosis-the journey of TF through NETs Vassilopoulos D, Monestier M, Y! Formation including circulating levels of serum tissue factor to neutrophil extracellular traps a predictor of venous!, McDonald B, Henke PK, et al B.A., Murthy, P.,,... For venous hydroxychloroquine pancreatic cancer ( VTE ) in patients with metastatic pancreatic adenocarcinoma in. Through NETs NIS Elements 4.4, Tokyo, Japan ) factor in our murine models of pancreatic is...