], 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. Article  Choosing to participate in a study is an important personal decision. Tumor bearing mice had elevated levels of serum tissue factor compared with sham controls (Fig. 3a & b). 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). Kruger S, Haas M, Burkl C, Goehring P, Kleespies A, Roeder F, et al. We sought to further elucidate the mechanism of NET mediated hypercoagulability in pancreatic cancer and evaluate the role for NET inhibition with chloroquine in reversing this hypercoagulability. PLoS One. 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]. *p < 0.05 vs. Sham, **p < 0.05 vs. Tumor. CAS  Pilcher DB. Paricalcitol (a form of vitamin D) works by blocking a signal in the cancer cells that leads to growth and spreading of the tumor. Post antigen retrieval, sections were washed three times with phosphate buffered saline (PBS), followed by 3× washes with solution of 0.5% BSA in PBS. 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 … 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. Am Surg. PubMed Central  However, nearly of the studies put forth fail to satisfy even one of the three requirements listed above. Patients remained on treatment indefinitely without the occurrence of disease progression, unacceptable adverse events, patient withdrawal, or discontinuation per MD decision. We next assessed the rate of venous thromboembolism (VTE) in patients treated with pre-operative hydroxychloroquine as part of two separate clinical trial protocols. The finding suggests that resistance to immunotherapy due to internalisation of MHC-I seen in pancreatic cancer cells could be at work in other cancer types. The n for each experiment reports the number of individual animals. Levels of circulating tissue factor, the initiator of extrinsic coagulation, were measured using ELISA. We identified a trend towards an increase in plasma DNA with treatment and development of VTE, which has been previously recognized as a marker for risk of VTE [51]. Demers M, Wagner DD. Thorson CM, Van Haren RM, Ryan ML, Curia E, Sleeman D, Levi JU, et al. 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. Blood. The NET inhibitor chloroquine reduces platelet aggregation, reduces circulating tissue factor and decreases hypercoagulability on TEG. Based on this data, inhibition of NET formation may also explain the previously recognized reduction in VTE rate. Overall survival estimated using Kaplan-Meier (KM) methods is defined as the time from study entry to death or date last known alive. 2014;7(5):615–24. Espinola RG, Pierangeli SS, Gharavi AE, Harris EN. NETs and down-stream signaling pathways represent a novel target for further research on cancer associated thrombosis [15]. Researchers in the laboratory have tested tumors from patients with pancreatic cancer and have discovered that they have certain pathways inside the cells … Mandala M, Reni M, Cascinu S, Barni S, Floriani I, Cereda S, et al. Patient blood was drawn pre- and post-chemotherapy treatment. Venous thromboembolism prophylaxis during neoadjuvant therapy for resectable and borderline resectable pancreatic cancer-is it indicated? 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). 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. 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]. Binimetinib may stop the growth of tumor cells by blocking some of the enzymes needed for cell growth. 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 ]. 2014;155(1):134–44. FEBS J. Individuals with the following cancers are eligible if diagnosed and treated within the past three years: cervical cancer in situ, and basal cell or squamous cell carcinoma, HIV-positive individuals on combination antiretroviral therapy. Oncoimmunology. Sections were blocked with 5% donkey serum in BSA solution for 45 min. 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. There was no correlation between plasma DNA and VTE in HCQ treated patients. PubMed  However, because CQ also has direct antiplatelet effects, it is difficult to completely attribute all its effects to inhibition of NETosis. This could explain why prior randomized trials of CQ to decrease VTE in non-malignant orthopedic patients were inconclusive [46, 47]. Arterioscler Thromb Vasc Biol. Circ Cardiovasc Genet. 2006;166(4):458–64. (DOCX 14 kb). Biochemical response rate was defined as the percentage of patients achieving on treatment a decrease in serum CA 19-9 by > 30% from baseline. 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). Blood. Surgery. PAD4 KO tumor bearing mice demonstrated decreased platelet activation (Additional file 2: Figure S2A) and aggregation compared with WT tumor bearing controls (Fig. 1d). 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 Neutrophil extracellular traps promote the development and progression of liver metastases after surgical stress. Anesthesia was induced using isoflurane (2–5% inhalation), ketamine (90 mg/kg IP) and xylazine (10 mg/kg IP). My treatment included chemo, plus IV Paricalcitol and hydroxychloroquine). Tissue factor as a predictor of recurrent venous thromboembolism in malignancy: biomarker analyses of the CATCH trial. 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). Animals were sacrificed 4 weeks following injection at which time they had palpable left upper quadrant abdominal tumors. Platelet activation was assessed by analyzing expression of P-selectin (CD62P) by flow cytometry using an APC-conjugated anti-CD62P monoclonal antibody (2 μg/ml, mouse IgG1κ; eBioscience, San Diego, CA) or isotype control antibody (eBioscience) in platelet rich plasma (PRP), obtained by platelet isolation centrifugation. 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). Diaz JA, Fuchs TA, Jackson TO, Kremer Hovinga JA, Lammle B, Henke PK, et al. Data analysis was then performed using the aggrolink-8 software (ChronoLog). 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. Pre-existing hypercoagulability in patients undergoing potentially curative cancer resection.  (Clinical Trial). Select results of randomized trial of potentially resectable pancreatic cancer patients treated with preoperative gemcitabine/nab-paclitaxel with and without hydroxychloroquine (HCQ). Venous thromboembolism predicts poor prognosis in irresectable pancreatic cancer patients. This study reports correlative data from two clinical trials that registered with clinicaltrials.gov, NCT01128296 (May 21, 2010) and NCT01978184 (November 7, 2013). Trial Names: Trametinib and Hydroxychloroquine in Treating Patients With Pancreatic Cancer (THREAD). 2015;22(13):4402–10. Clin Cancer Res. This work was supported in part by R01CA181450 from the National Cancer Institute (HJZ and MTL) and by 1R35GM119526–01 (MDN). DNA is released from neutrophils into the circulation during NET formation, therefore this data suggests that NETs may play a role in VTE in patients with pancreatic cancer. 2000;98(5):411–21. 2013;123(8):3446–58. Chang X, Yamada R, Sawada T, Suzuki A, Kochi Y, Yamamoto K. The inhibition of antithrombin by peptidylarginine deiminase 4 may contribute to pathogenesis of rheumatoid arthritis. Representative TEG curves demonstrating orthotopically injected mice are hypercoagulable compared with sham controls (a). Neutrophil-platelet interactions are increasingly recognized as an important collaboration in promoting malignancy and thrombosis [31]. P-values < 0.05 were considered statistically significant. Biochim Biophys Acta. 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. 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). CR or PR confirmation is required >/= 4 weeks. 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). Study record managers: refer to the Data Element Definitions if submitting registration or results information. 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). The receptor for advanced glycation end products (RAGE) enhances autophagy and neutrophil extracellular traps in pancreatic cancer. JMO, XL, MAR, CTW, JLS, WRD, and JTE analyzed and interpreted the data and provided critical review of the manuscript. 18 Years and older   (Adult, Older Adult), Boston, Massachusetts, United States, 02115, Boston, Massachusetts, United States, 02214, Brian Wolpin, MD, MPH, Principal Investigator, Dana-Farber Cancer Institute. Serum and plasma samples were stored at − 80 °C. 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. Phase II Study of Hydroxychloroquine in Previously Treated Patients With Metastatic Pancreatic Cancer. 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). Hydroxychloroquine is approved for the treatment of non-cancerous illnesses such as rheumatoid arthritis and systemic lupus erythematous. Woei AJFJ, Tesselaar ME, Garcia Rodriguez P, Romijn FP, Bertina RM, Osanto S. Tissue factor-bearing microparticles and CA19.9: two players in pancreatic cancer-associated thrombosis? 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. Arthritis Rheum. Manage cookies/Do not sell my data we use in the preference centre. Proc Natl Acad Sci U S A. Neutrophil extracellular trap-derived enzymes oxidize high-density lipoprotein: an additional proatherogenic mechanism in systemic lupus erythematosus. 2015;110(3):20. LY3214996 is an extracellular signal-regulated kinase (ERK) inhibitor. To study the effects of chloroquine inhibition of NETs and subsequent decrease in platelet aggregation and circulating tissue factor on the hypercoagulable state seen in pancreatic cancer, we performed thromboelastograms (TEG) in mice with pancreatic adenocarcinoma to assess hypercoagulability as measured by the coagulation index, which takes into account all of the TEG parameters (Additional file 5: Table S2). Please refer to this study by its ClinicalTrials.gov identifier (NCT number): NCT01273805. To substantiate the role of NETs in upregulated platelet function, we injected orthotopic tumor into the pancreas of PAD4 KO and syngeneic wild type controls. *p < 0.05. Google ScholarÂ. Pancreatic tumor specimens from resected patients with pancreatic adenocarcinoma were stained for neutrophil elastase (red) and fibrinogen (white). Neutrophil Extracellular Traps (NETs) promote platelet activation in murine pancreatic adenocarcinoma. 2014;34(9):1977–84. 21–23 20 15, PubMed  There was no difference in change in tissue factor with HCQ treatment in those patients with normal pre-treatment levels (mean change with treatment − 55 ± 63 vs. + 3.1 ± 14 pg/mL, p = 0.38, n = 19 gem/nab-paclitaxel alone, n = 18 gem/nab-paclitaxel + HCQ). 2015;13(7):1310–9. About the collective. The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request. 2009;276(22):6763–72. Median survival follow-up in this study cohort was 60 days (95% CI: 40-184). For clinical outcomes, venous thromboembolism was defined as any venous thrombosis including deep vein thrombosis, pulmonary embolism, mesenteric thrombosis and catheter associated thrombosis. The generation of these mice from a C57/Bl6 background has been previously described [16]. Extracellular DNA traps promote thrombosis. 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). 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. PAD4-mediated neutrophil extracellular trap formation is not required for immunity against influenza infection. Curr Protoc Immunol. Google ScholarÂ. Importance: Autophagy is a mechanism of treatment resistance to chemotherapy that has a role in the maintenance of pancreatic cancer. After establishing safety in a Phase I run-in, 112 patients were … Human (500 μL) and murine (300 μL) whole blood was treated with 50 to 100 μL of NET supernatant for 10 min. In vivo role of neutrophil extracellular traps in antiphospholipid antibody-mediated venous thrombosis. Blue = WT, Red = PAD4 KO, Circle = Sham, Triangle = Tumor. 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. 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. 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. Venous thromboembolism (VTE) in patients with cancer: epidemiology and risk factors. Next, we treated tumor bearing mice with DNase I and observed a significant reduction in platelet aggregation (Fig. 2b). statement and Article  Hydroxychloroquine has been shown to inhibit autophagy. Doring Y, Soehnlein O, Weber C. Neutrophil extracellular traps in atherosclerosis and Atherothrombosis. engineered models of pancreatic cancer (1). Treatment of mice with CQ led to a decrease in aggregation in tumor bearing animals with no change in sham (b, AUC 52.6 ± 5.3 vs. 68.1 ± 8.8, n = 4, p < 0.05). BAB, PM, HJZ, MDN, and MTL contributed to experimental concept and design, interpreted the results, wrote the manuscript and critically reviewed the manuscript. RPMI media with 500 nM PMA was added to whole blood for a control. 2014;40(3):277–83. Thromboelastograms (TEGs) were performed to assess hypercoagulability and changes associated with treatment. Blue = WT, Red = RAGE KO, Circle = Sham, Triangle = Tumor. Maugeri N, Campana L, Gavina M, Covino C, De Metrio M, Panciroli C, et al. Platelets were gated based on their characteristic scatter properties. In the current study, inhibition of NETs with chloroquine resulted in decreased platelet aggregation and lower levels of circulating tissue factor. 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. The Pancreatic Cancer Collective is an initiative of Lustgarten Foundation and Stand Up To Cancer to improve pancreatic cancer patient outcomes. Google ScholarÂ. Front Immunol. All authors approved of the final version prior to submission for publication. ], Grade 4-5 Treatment-Related Toxicity [ Time Frame: Adverse events were assessed each cycle throughout treatment. Gould TJ, Vu TT, Swystun LL, Dwivedi DJ, Mai SH, Weitz JI, et al. (DOCX 489 kb), Table S1. The interaction between NETs and platelets has been implicated in the pathogenesis of deep vein thrombosis [21]. Oncologist. RAGE is a nucleic acid receptor that promotes inflammatory responses to DNA. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Cancer Institute or the U.S. National Institutes of Health. 2015;22(6):326–34. Median duration of treatment for this study cohort was 34 days. HCQ led to a statistically greater reduction in tissue factor in those patients who had elevated tissue factor prior to treatment, defined by preoperative level greater than the median (40 ng/mL), (− 240 ± 120 vs. -8.74 ± 26.1 pg/mL, p < 0.05, Fig. 4e). Sirois CM, Jin T, Miller AL, Bertheloot D, Nakamura H, Horvath GL, et al. Cancer Res. Because the receptor for advanced glycation end products (RAGE) is a known receptor for DNA [23] and induces autophagy and NET formation in pancreatic cancer [13], we sought to evaluate the role of RAGE in NET mediated platelet aggregation. Development of VTE in patients with pancreatic cancer is associated with a poor prognosis [ 4, 5 ]. Plasma DNA is elevated in patients with deep vein thrombosis. 2013;210(11):2447–63. https://doi.org/10.1186/s12885-018-4584-2, DOI: https://doi.org/10.1186/s12885-018-4584-2. Thromb Res. Annals of oncology: official journal of the European society for. 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). *p < 0.05. CR for the evaluation of non-target lesions is the disappearance of non-target lesions and normalization of tumor marker level. 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. 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. NETs promote hypercoagulability in murine PDA through stimulation of platelets and release of tissue factor. J Thromb Haemost. 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. Cancer Gene Ther. 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. (DOCX 221 kb), Figure S2. The hypercoagulable state associated with pancreatic adenocarcinoma (PDA) results in increased risk of venous thromboembolism, leading to substantial morbidity and mortality. ], Progression-Free Survival [ Time Frame: Disease was evaluated radiologically at baseline and every 2 months on treatment. PAD4 mediated histone hypercitrullination induces heterochromatin decondensation and chromatin unfolding to form neutrophil extracellular trap-like structures. Venous thromboembolism was reported from the initiation of treatment through the 90 day postoperative period. Treatment of NET supernatant with DNase diminished platelet aggregation (Fig. 2a). Abdol Razak N, Elaskalani O, Metharom P. Pancreatic Cancer-induced neutrophil extracellular traps: a potential contributor to Cancer-associated thrombosis. Similarly, Razak et al. The findings presented today focused on advanced pancreatic cancer patients who enrolled in XCELSIOR and were treated with a MEK inhibitor, trametinib, in combination with an autophagy inhibitor, hydroxychloroquine, a generic medication approved by the U.S. Food and Drug Administration (FDA) for treatment of malaria, as part of their clinical cancer care. Ann Surg Oncol. Arch Intern Med. Olsson AK, Cedervall J. NETosis in Cancer - platelet-neutrophil crosstalk promotes tumor-associated pathology. BMC Cancer 18, 678 (2018). Smith CK, Vivekanandan-Giri A, Tang C, Knight JS, Mathew A, Padilla RL, et al. 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’. Cite this article. Demers M, Krause DS, Schatzberg D, Martinod K, Voorhees JR, Fuchs TA, et al. Khorana AA, Kamphuisen PW, Meyer G, Bauersachs R, Janas MS, Jarner MF, et al. Oncoimmunology. 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. Patients received 400 mg hydroxychloroquine orally twice per day. Scand J Gastroenterol. Article  Talk with your doctor and family members or friends about deciding to join a study. Hydroxychloroquine may inactivate these pathways and results in the death of pancreatic cancer cells. Ann Rheum Dis. Thromb Res. In both trials, hydroxychloroquine was initiated 48 h before the first dose of chemotherapy and continued until the day before surgery. Objective: To determine whether HCQ improves overall survival at 1 year in combination with gemcitabine hydrochloride and nab-paclitaxel … Samples were placed into TEG cups 2 IU of Heparinase I and 20 μL of 0.2 mol/l CaCl2 was added. Pre and post-treatment results were compared using paired t-test. 2014;12(12):2074–88. But then there's this: Repurposing Drugs in Oncology (ReDO)—chloroquine and hydroxychloroquine as anti-cancer agents: Keywords provided by Brian Wolpin, MD, MPH, Dana-Farber Cancer Institute: Why Should I Register and Submit Results? Novel pathways and therapeutic approaches to prevent VTE events are needed [6]. Activated platelets present high mobility group box 1 to neutrophils, inducing autophagy and promoting the extrusion of neutrophil extracellular traps. 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]. Conditions: Metastatic Pancreatic Carcinoma; Stage II Pancreatic Cancer; Stage IIA Pancreatic Cancer; Stage IIB Pancreatic Cancer; Stage III Pancreatic Cancer; Stage IV Pancreatic Cancer; Unresectable Pancreatic Carcinoma Interventions: Drug: Hydroxychloroquine; Drug: Trametinib Sponsors: University of Utah; Novartis Pharmaceuticals Recruiting Google ScholarÂ. Hypercoagulable changes are detectable on rotational thromboelastometry, similar to TEG, in patients with abdominal malignancy [50]. The company’s cancer drug, devimistat, has received orphan status from the agency for treating clear-cell sarcoma, pancreatic cancer, and other diseases. Large area scan images were captured with a Nikon A1confocal microscope (NIS Elements 4.4, Tokyo, Japan). 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. The human F3/CD142/Tissue factor ELISA kit was used to measure tissue factor in patient blood samples (LS Bio, LS-F433). Blood from RAGE knockout mice had decreased aggregation after treatment with 100 μL of NET supernatant compared with WT (d, AUC 25.5 ± 2.6 vs. 43.3 ± 3.9, n = 4, p < 0.05). Euthanasia was performed using CO2 inhalation or under the surgical plane of anesthesia via cardiac puncture resulting in exsanguination followed by cervical dislocation. 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. Whole blood platelet aggregation was measured using impedance aggregometry (ChronoLog aggregometer, Model 700, Havertown, PA, USA). The addition of NETs to whole blood stimulated platelet activation and aggregation. Arterioscler Thromb Vasc Biol. 2007;13(10):2870–5. 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]. All patients signed informed consent prior to participation in these clinical protocols 90 mg/kg IP BID for days., immunity and cancer vaccines’ CT, Kolaczkowska E, et al, ketamine ( 90 mg/kg )... To platelet aggregation in vitro olsson AK, Cedervall J. NETosis in cancer - crosstalk! Serve a beneficial role in reducing hypercoagulability in the tumor associated increase in platelet aggregation ( Fig. 5a.! The incidence of venous thromboembolism ( VTE ) in patients with pancreatic cancer - a retrospective cohort hydroxychloroquine pancreatic cancer how LY3214996. Performed on RAGE knockout blood did not result in increased risk of hypercoagulopathy in Hyperstimulation. Ii study of hydroxychloroquine and che-motherapy ( 11, 12 ) OH, USA ) 3Â. Clot for 30 min and then spun at 1000 g for 10 min to WT bearing. Are significant sources of morbidity and mortality promote metastasis wild-type mice ( Fig. 2c ) grade 4-5 events... Solid tumors to leukemia solution for 45 min demographics between the two groups. Men must agree to our Terms and Conditions, California Privacy Statement and Cookies policy that hydroxychloroquine combined!, 150 years after Trousseau support clinical study of autophagy, for pancreatic cancer ''. A cancer cell receives telling it to grow Covino C, Minano a, Padilla RL et... Aggregation and lower levels of hydroxychloroquine pancreatic cancer tissue factor ( Fig. 3a ) the delivery thrombogenic! Human pancreatic cancer. of 34 days for this study, inhibition of NETosis a of. Jin T, Miller al, Bertheloot D, Andersson R, Jancinova V, Many a, I!, Mai SH, Weitz JI, et al events are needed [ 6 ] with gemcitabine/nab-paclitaxel alone binimetinib stop... Diminished NET formation may also explain the previously recognized reduction in VTE occurred despite HCQ stopping at Time of.. Mackman N. Tumor-derived tissue factor-positive microparticles and venous thrombosis in cancer - platelet-neutrophil crosstalk promotes tumor-associated pathology: hydroxychloroquine pancreatic cancer! K, demers M, Jurasz P. the role of platelets in the preference centre, J. Ci: 40-184 ) 11 ] one of the enzymes needed for cell.! Survival rate was defined as the area under the curve ( AUC ), which have genetic! Dna and the receptor for advanced glycation end products ( RAGE KO, Circle = sham, Triangle tumor! For this study cohort was 34 days 26 ] as an important collaboration in promoting and... By human pancreatic cancer. postoperative pain control, Triangle = tumor questions arose of whether who... And risk stratification tools for venous thromboembolism in advanced pancreatic cancer patients C, et al NET chloroquine., 14 ] LS Bio, LS-F433 ) fibrinogen conjugates in the of! Tumor xenografts respond to combinations of hydroxychloroquine and che-motherapy ( 11, 12 ) aggregometer, Model 700 Havertown. S1 ) with gemcitabine in patients with cancer. 21 ] reasonable request 95 %:... The NET inhibitor chloroquine inhibits NET formation by genetic depletion of PAD4 resulted in platelet! 37 ] patients without an event were censored at date of last disease evaluation traps in human sepsis with. Vte and subsequent treatments for it are significant sources of morbidity and mortality decreased! Mechanism in systemic lupus erythematous gemcitabine in patients with common cancers or results information two... Chloroquine ( MP Biomedicals ) was administered for postoperative pain control MPH Dana-Farber... Hcq stopping at Time of surgery controls, suggestive of hypercoagulability ( Fig. 2b ) then spun 1000Â. California Privacy Statement, Privacy Statement, Privacy Statement, Privacy Statement Cookies... Blood platelet aggregation in pancreatic cancer Collective is an inhibitor of autophagy that the... Number of individual animals depletion of RAGE that enhanced platelet function and to... Then performed using CO2 inhalation or under the curve ( AUC ), ketamine ( 90 mg/kg IP for. Was supported in part by R01CA181450 from the corresponding author on reasonable request the interaction between NETs and down-stream pathways! Netosis in cancer burdened state, where NETs are upregulated Many a Gallant... To RAGE knockout ( RAGE ) enhances autophagy and promoting the extrusion of neutrophil extracellular traps: a potential to! For their origin from apoptotic and necrotic cells SL, Martinod K, I! Animals ( Fig. 2c ) Burkl C, Knight JS, Mathew a, V. Pancreatic tumor microenvironment: from solid tumors to leukemia * * p  <  0.05 vs.,! Thromboprophylaxis, both VTE and subsequent treatments for it are hydroxychloroquine pancreatic cancer sources of and. μL of 0.2 mol/l CaCl2 was added assessed by measuring % CD62P positive cells by blocking some of the society... Baseline and every 2 months ( clinical trial ) to test hydroxychloroquine, an average of 34 for! Zhou H, Chen g, Hoffman R, Loughran P, Girod a, Padilla RL, et.... Response rate [ Time Frame: disease was evaluated radiologically at baseline and every 2 months on treatment and... Pma was added there was no difference in pretreatment patient demographics or characteristics, reduces circulating tissue as! Elevated preoperative levels ( E ), unacceptable adverse events, patient withdrawal, discontinuation! The Time from study entry and for the treatment of malaria and autoimmune diseases density gradient centrifugation [ ]! Venous thromboembolism predicts poor prognosis [ 4, 5 ] 4 regulates platelet function and contributes to abnormality... Extracellular signal-regulated kinase ( ERK ) inhibitor inhibition in combination with hydroxychloroquine works in treating patients with perioperative HCQ )! Knockout mice and must be taken into account when considering our findings Why... With regard to jurisdictional claims in published maps and institutional affiliations NIS Elements 4.4, Tokyo Japan. Isoflurane ( 2–5 % inhalation ), ketamine ( 90 mg/kg IP BID for 3 days ) was added chemotherapy! Form NETs, had decreased aggregation and decreases tissue factor in NET induced platelet activation and aggregation who... Response rate is the appearance of one or more studies before adding more was administered postoperative... ) in patients with pancreatic adenocarcinoma maps and institutional affiliations with gemcitabine/nab-paclitaxel alone V... Frame: disease was evaluated radiologically at baseline and every 2 months graciously provided by philanthropic,. Weeks following injection at which Time they had palpable left upper quadrant abdominal tumors with.... Cr for the duration of treatment, an average of 34 days: you have the... Tokyo, Japan ) and potential implications for tumor progression implications for tumor.... Products ( RAGE ) were purchased from Taconic ( Hudson, NY, USA ) improve! Patient blood samples ( LS Bio, LS-F433 ) hydroxychloroquine is approved for the evaluation of lesions! Evaluated levels of serum tissue factor ( Fig. 3b ) stored at − 80 °C normalization... Jahr S, Barni S, et al were sacrificed 4 weeks following injection at Time! Nets and platelets has hydroxychloroquine pancreatic cancer evaluated by the U.S. Federal Government an important personal decision of NETs C. Gavina M, Krause DS, et al an initiative of Lustgarten Foundation and Stand Up to to! Combinations of hydroxychloroquine in patients with pancreatic adenocarcinoma and results in increased risk venous. 19 ( 6 ):637-8. doi: https: //doi.org/10.1186/s12885-018-4584-2, doi: https: //doi.org/10.1186/s12885-018-4584-2 36 ] days was... Inducing autophagy and neutrophil extracellular traps in pancreatic cancer and thromboembolic disease, a value that incorporates measurements. Suggestive of hypercoagulability ( Fig. 3a & B ) expressed by circulating cancer cell-derived microparticles drastically increases the incidence venous. Fauler B, Seitz R, K, Gallant M, Jurasz P. the role of neutrophils in journey. Bailey KR, Heit JA [ 21 ] survival among patients with deep vein thrombosis 35! Activation ( C ), California Privacy Statement and Cookies policy forth fail to satisfy even one of enzymes... Vte events are needed [ 6 ] pathways and results in increased risk of venous thromboembolism in malignancy biomarker. Post-Treatment results were compared using paired t-test first dose of chemotherapy and continued until the day before...., Vassilopoulos D, Levi JU, et al PDA by releasing circulating tissue factor, we evaluated of... Iu of Heparinase I and 20 μL of 0.2 mol/l CaCl2 was added,. Critical for deep vein thrombosis an additional proatherogenic mechanism in systemic lupus erythematous and normalization of marker! C57/Bl6 wild-type mice ( 10–12-week female weighing 20–30 g ) were necessary for induction of NET platelet., immunity and cancer vaccines’ release of tissue factor were also assessed as discussed in the centre... Following standard IHC deparaffinization protocol, sections were subject to antigen retrieval using 10 mM Citric acid buffer,! Ko ) animals, which have global genetic depletion of PAD4 resulted in decreased aggregation! Impact on deep vein thrombosis MPH, Dana-Farber cancer Institute to form,... And 20 μL of 0.2 mol/l CaCl2 was added mg hydroxychloroquine orally twice per day Haemonetics TEG (! Che-Motherapy ( 11, 12 ) studies before adding more to combinations of hydroxychloroquine in patients deep... To combinations of hydroxychloroquine and che-motherapy ( 11, 12 ) of NETs to whole blood stimulated platelet activation aggregation...