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Thromb Haemost 2018; 118(12): 2064-2073
DOI: 10.1055/s-0038-1675788
Coagulation and Fibrinolysis
Georg Thieme Verlag KG Stuttgart · New York

Increased Neutrophil Activation and Plasma DNA Levels in Patients with Pre-Eclampsia

Yae Hu   *
1   Cyrus Tang Hematology Center, Collaborative Innovation Center of Hematology, State Key Laboratory of Radiation Medicine and Prevention, Soochow University, Suzhou, China
2   Department of Pathophysiology, Medical School of Nantong University, Nantong University, Nantong, China
,
Hui Li   *
3   Central Laboratory, Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou Municipal Hospital, Suzhou, China
,
Ruhong Yan
1   Cyrus Tang Hematology Center, Collaborative Innovation Center of Hematology, State Key Laboratory of Radiation Medicine and Prevention, Soochow University, Suzhou, China
,
Can Wang
1   Cyrus Tang Hematology Center, Collaborative Innovation Center of Hematology, State Key Laboratory of Radiation Medicine and Prevention, Soochow University, Suzhou, China
,
Yun Wang
4   Department of Obstetrics and Gynecology, Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou Municipal Hospital, Suzhou, China
,
Ce Zhang
1   Cyrus Tang Hematology Center, Collaborative Innovation Center of Hematology, State Key Laboratory of Radiation Medicine and Prevention, Soochow University, Suzhou, China
,
Meng Liu
1   Cyrus Tang Hematology Center, Collaborative Innovation Center of Hematology, State Key Laboratory of Radiation Medicine and Prevention, Soochow University, Suzhou, China
,
Tiantian Zhou
1   Cyrus Tang Hematology Center, Collaborative Innovation Center of Hematology, State Key Laboratory of Radiation Medicine and Prevention, Soochow University, Suzhou, China
,
Weipei Zhu
5   Department of Obstetrics and Gynecology, The Second Affiliated Hospital of Soochow University, Soochow University, Suzhou, China
,
Hong Zhang
5   Department of Obstetrics and Gynecology, The Second Affiliated Hospital of Soochow University, Soochow University, Suzhou, China
,
Ningzheng Dong
1   Cyrus Tang Hematology Center, Collaborative Innovation Center of Hematology, State Key Laboratory of Radiation Medicine and Prevention, Soochow University, Suzhou, China
6   MOH Key Laboratory of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, Medical College, The First Affiliated Hospital of Soochow University, Soochow University, Suzhou, China
7   Jiangsu Key Laboratory of Preventive and Translational Medicine for Geriatric Diseases, Soochow University, Suzhou, China
,
Qingyu Wu
1   Cyrus Tang Hematology Center, Collaborative Innovation Center of Hematology, State Key Laboratory of Radiation Medicine and Prevention, Soochow University, Suzhou, China
7   Jiangsu Key Laboratory of Preventive and Translational Medicine for Geriatric Diseases, Soochow University, Suzhou, China
8   Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio, United States
› Author Affiliations Funding This work was supported in part by grants from the National Science Foundation of China (81671485, 81503304, 81570457 and 91639116), the National Basic Research Program of China (2015CB943302), the Priority Academic Program Development of Jiangsu Higher Education Institutes and the NIH (HD093727 and HL126697).
Further Information

Publication History

04 June 2018

06 October 2018

Publication Date:
19 November 2018 (online)

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Abstract

Pre-eclampsia (PE) is a chronic inflammatory disease in pregnancy, which is associated with enhanced blood coagulation and high thrombotic risk. To date, the mechanisms underlying such an association are not fully understood. Previous studies reported high levels of plasma deoxyribonucleic acid (DNA) in PE women, but the cellular source of the circulation DNA remains unknown. In this study, we tested the hypothesis that activated neutrophils undergoing cell death, also called NETosis, may be responsible for the elevated plasma DNA levels in PE women. We analysed plasma samples from non-pregnant, normal pregnant and PE women and found high levels of double-stranded DNA, myeloperoxidase (an abundant neutrophil granular enzyme) and histones (the major nucleosome proteins) in PE-derived samples, indicating increased NETosis in the maternal circulation. The high plasma DNA levels positively correlated with enhanced blood coagulation in PE women. When isolated neutrophils from normal individuals were incubated with PE-derived plasma, an elevated NETosis-stimulating activity was detected. Further experiments showed that endothelial micro-particles, but not soluble proteins, in the plasma were primarily responsible for the NETosis-stimulating activity in PE women. These results indicate that circulating micro-particles from damaged maternal endothelium are a potent stimulator for neutrophil activation and NETosis in PE women. Given the pro-coagulant and pro-thrombotic nature of granular and nuclear contents from neutrophils, enhanced systemic NETosis may represent an important mechanism underlying the hyper-coagulability and increased thrombotic risk in PE.

Keywords

hyper-coagulability - hypertension - micro-particles - NETosis - neutrophils - Pre-eclampsia

* These authors contributed equally to this work.


Supplementary Material

 

  • References

  • 1 Chaiworapongsa T, Chaemsaithong P, Yeo L, Romero R. Pre-eclampsia part 1: current understanding of its pathophysiology. Nat Rev Nephrol 2014; 10 (08) 466-480
    CrossrefPubMedGoogle Scholar
  • 2 Cadroy Y, Grandjean H, Pichon J. , et al. Evaluation of six markers of haemostatic system in normal pregnancy and pregnancy complicated by hypertension or pre-eclampsia. Br J Obstet Gynaecol 1993; 100 (05) 416-420
    CrossrefPubMedGoogle Scholar
  • 3 Chambers JC, Fusi L, Malik IS, Haskard DO, De Swiet M, Kooner JS. Association of maternal endothelial dysfunction with preeclampsia. JAMA 2001; 285 (12) 1607-1612
    Google Scholar
  • 4 George JN. The association of pregnancy with thrombotic thrombocytopenic purpura-hemolytic uremic syndrome. Curr Opin Hematol 2003; 10 (05) 339-344
    CrossrefPubMedGoogle Scholar
  • 5 Roberts JM. Pathophysiology of ischemic placental disease. Semin Perinatol 2014; 38 (03) 139-145
    CrossrefPubMedGoogle Scholar
  • 6 Pijnenborg R, Vercruysse L, Hanssens M. The uterine spiral arteries in human pregnancy: facts and controversies. Placenta 2006; 27 (9-10): 939-958
    CrossrefPubMedGoogle Scholar
  • 7 Redman CW, Sargent IL. Latest advances in understanding preeclampsia. Science 2005; 308 (5728): 1592-1594
    CrossrefPubMedGoogle Scholar
  • 8 Cui Y, Wang W, Dong N. , et al. Role of corin in trophoblast invasion and uterine spiral artery remodelling in pregnancy. Nature 2012; 484 (7393): 246-250
    PubMedGoogle Scholar
  • 9 Dong N, Zhou T, Zhang Y. , et al. Corin mutations K317E and S472G from preeclamptic patients alter zymogen activation and cell surface targeting. J Biol Chem 2014; 289 (25) 17909-17916
    CrossrefPubMedGoogle Scholar
  • 10 Red-Horse K, Zhou Y, Genbacev O. , et al. Trophoblast differentiation during embryo implantation and formation of the maternal-fetal interface. J Clin Invest 2004; 114 (06) 744-754
    CrossrefPubMedGoogle Scholar
  • 11 Zhou Y, Wu Q. Role of corin and atrial natriuretic peptide in preeclampsia. Placenta 2013; 34 (02) 89-94
    CrossrefPubMedGoogle Scholar
  • 12 Hu Y, Yan R, Zhang C. , et al. High-mobility group box 1 from hypoxic trophoblasts promotes endothelial microparticle production and thrombophilia in preeclampsia. Arterioscler Thromb Vasc Biol 2018; 38 (06) 1381-1391
    CrossrefPubMedGoogle Scholar
  • 13 Harris HE, Andersson U, Pisetsky DS. HMGB1: a multifunctional alarmin driving autoimmune and inflammatory disease. Nat Rev Rheumatol 2012; 8 (04) 195-202
    CrossrefPubMedGoogle Scholar
  • 14 Fuchs TA, Abed U, Goosmann C. , et al. Novel cell death program leads to neutrophil extracellular traps. J Cell Biol 2007; 176 (02) 231-241
    CrossrefPubMedGoogle Scholar
  • 15 Caudrillier A, Kessenbrock K, Gilliss BM. , et al. Platelets induce neutrophil extracellular traps in transfusion-related acute lung injury. J Clin Invest 2012; 122 (07) 2661-2671
    CrossrefPubMedGoogle Scholar
  • 16 Cools-Lartigue J, Spicer J, McDonald B. , et al. Neutrophil extracellular traps sequester circulating tumor cells and promote metastasis. J Clin Invest 2013; 123: 3446-3458
    CrossrefPubMedGoogle Scholar
  • 17 Hakkim A, Fürnrohr BG, Amann K. , et al. Impairment of neutrophil extracellular trap degradation is associated with lupus nephritis. Proc Natl Acad Sci U S A 2010; 107 (21) 9813-9818
    CrossrefPubMedGoogle Scholar
  • 18 Kessenbrock K, Krumbholz M, Schönermarck U. , et al. Netting neutrophils in autoimmune small-vessel vasculitis. Nat Med 2009; 15 (06) 623-625
    CrossrefPubMedGoogle Scholar
  • 19 Wang Y, Gu Y, Philibert L, Lucas MJ. Neutrophil activation induced by placental factors in normal and pre-eclamptic pregnancies in vitro. Placenta 2001; 22 (06) 560-565
    CrossrefPubMedGoogle Scholar
  • 20 Altincicek B, Stötzel S, Wygrecka M, Preissner KT, Vilcinskas A. Host-derived extracellular nucleic acids enhance innate immune responses, induce coagulation, and prolong survival upon infection in insects. J Immunol 2008; 181 (04) 2705-2712
    CrossrefPubMedGoogle Scholar
  • 21 Ammollo CT, Semeraro F, Xu J, Esmon NL, Esmon CT. Extracellular histones increase plasma thrombin generation by impairing thrombomodulin-dependent protein C activation. J Thromb Haemost 2011; 9 (09) 1795-1803
    CrossrefPubMedGoogle Scholar
  • 22 Fuchs TA, Brill A, Duerschmied D. , et al. Extracellular DNA traps promote thrombosis. Proc Natl Acad Sci U S A 2010; 107 (36) 15880-15885
    CrossrefPubMedGoogle Scholar
  • 23 Massberg S, Grahl L, von Bruehl ML. , et al. Reciprocal coupling of coagulation and innate immunity via neutrophil serine proteases. Nat Med 2010; 16 (08) 887-896
    CrossrefPubMedGoogle Scholar
  • 24 Xu J, Zhang X, Pelayo R. , et al. Extracellular histones are major mediators of death in sepsis. Nat Med 2009; 15 (11) 1318-1321
    CrossrefPubMedGoogle Scholar
  • 25 Barden A, Graham D, Beilin LJ. , et al. Neutrophil CD11B expression and neutrophil activation in pre-eclampsia. Clin Sci (Lond) 1997; 92 (01) 37-44
    CrossrefPubMedGoogle Scholar
  • 26 Gervasi MT, Chaiworapongsa T, Pacora P. , et al. Phenotypic and metabolic characteristics of monocytes and granulocytes in preeclampsia. Am J Obstet Gynecol 2001; 185 (04) 792-797
    CrossrefPubMedGoogle Scholar
  • 27 Sacks GP, Studena K, Sargent K, Redman CW. Normal pregnancy and preeclampsia both produce inflammatory changes in peripheral blood leukocytes akin to those of sepsis. Am J Obstet Gynecol 1998; 179 (01) 80-86
    CrossrefPubMedGoogle Scholar
  • 28 Tsukimori K, Maeda H, Ishida K, Nagata H, Koyanagi T, Nakano H. The superoxide generation of neutrophils in normal and preeclamptic pregnancies. Obstet Gynecol 1993; 81 (04) 536-540
    PubMedGoogle Scholar
  • 29 Mellembakken JR, Aukrust P, Olafsen MK, Ueland T, Hestdal K, Videm V. Activation of leukocytes during the uteroplacental passage in preeclampsia. Hypertension 2002; 39 (01) 155-160
    CrossrefPubMedGoogle Scholar
  • 30 Gupta A, Hasler P, Gebhardt S, Holzgreve W, Hahn S. Occurrence of neutrophil extracellular DNA traps (NETs) in pre-eclampsia: a link with elevated levels of cell-free DNA?. Ann N Y Acad Sci 2006; 1075: 118-122
    CrossrefPubMedGoogle Scholar
  • 31 Gupta AK, Hasler P, Holzgreve W, Gebhardt S, Hahn S. Induction of neutrophil extracellular DNA lattices by placental microparticles and IL-8 and their presence in preeclampsia. Hum Immunol 2005; 66 (11) 1146-1154
    CrossrefPubMedGoogle Scholar
  • 32 Hahn S, Giaglis S, Hoesli I, Hasler P. Neutrophil NETs in reproduction: from infertility to preeclampsia and the possibility of fetal loss. Front Immunol 2012; 3: 362
    CrossrefPubMedGoogle Scholar
  • 33 Zhang Y, Zhou T, Niu Y. , et al. Identification and functional analysis of CORIN variants in hypertensive patients. Hum Mutat 2017; 38 (12) 1700-1710
    CrossrefPubMedGoogle Scholar
  • 34 Keshari RS, Jyoti A, Kumar S. , et al. Neutrophil extracellular traps contain mitochondrial as well as nuclear DNA and exhibit inflammatory potential. Cytometry A 2012; 81 (03) 238-247
    PubMedGoogle Scholar
  • 35 Farina A, Sekizawa A, Iwasaki M, Matsuoka R, Ichizuka K, Okai T. Total cell-free DNA (beta-globin gene) distribution in maternal plasma at the second trimester: a new prospective for preeclampsia screening. Prenat Diagn 2004; 24 (09) 722-726
    CrossrefPubMedGoogle Scholar
  • 36 Gänshirt D, Smeets FW, Dohr A. , et al. Enrichment of fetal nucleated red blood cells from the maternal circulation for prenatal diagnosis: experiences with triple density gradient and MACS based on more than 600 cases. Fetal Diagn Ther 1998; 13 (05) 276-286
    CrossrefPubMedGoogle Scholar
  • 37 Swinkels DW, de Kok JB, Hendriks JC, Wiegerinck E, Zusterzeel PL, Steegers EA. Hemolysis, elevated liver enzymes, and low platelet count (HELLP) syndrome as a complication of preeclampsia in pregnant women increases the amount of cell-free fetal and maternal DNA in maternal plasma and serum. Clin Chem 2002; 48 (04) 650-653
    CrossrefPubMedGoogle Scholar
  • 38 Wataganara T, Bianchi DW. Fetal cell-free nucleic acids in the maternal circulation: new clinical applications. Ann N Y Acad Sci 2004; 1022: 90-99
    CrossrefPubMedGoogle Scholar
  • 39 Zhong XY, Laivuori H, Livingston JC. , et al. Elevation of both maternal and fetal extracellular circulating deoxyribonucleic acid concentrations in the plasma of pregnant women with preeclampsia. Am J Obstet Gynecol 2001; 184 (03) 414-419
    CrossrefPubMedGoogle Scholar
  • 40 Harmon AC, Cornelius DC, Amaral LM. , et al. The role of inflammation in the pathology of preeclampsia. Clin Sci (Lond) 2016; 130 (06) 409-419
    CrossrefPubMedGoogle Scholar
  • 41 Nadeau-Vallée M, Obari D, Palacios J. , et al. Sterile inflammation and pregnancy complications: a review. Reproduction 2016; 152 (06) R277-R292
    CrossrefPubMedGoogle Scholar
  • 42 Redman CW, Sacks GP, Sargent IL. Preeclampsia: an excessive maternal inflammatory response to pregnancy. Am J Obstet Gynecol 1999; 180 (2 Pt 1): 499-506
    CrossrefPubMedGoogle Scholar
  • 43 Erpenbeck L, Chowdhury CS, Zsengellér ZK. , et al. PAD4 deficiency decreases inflammation and susceptibility to pregnancy loss in a mouse model. Biol Reprod 2016; 95 (06) 132
    CrossrefPubMedGoogle Scholar
  • 44 Gupta AK, Joshi MB, Philippova M. , et al. Activated endothelial cells induce neutrophil extracellular traps and are susceptible to NETosis-mediated cell death. FEBS Lett 2010; 584 (14) 3193-3197
    CrossrefPubMedGoogle Scholar
  • 45 Halim A, Kanayama N, El Maradny E, Maehara K, Bhuiyan AB, Terao T. Correlated plasma elastase and sera cytotoxicity in eclampsia. A possible role of endothelin-1 induced neutrophil activation in preeclampsia-eclampsia. Am J Hypertens 1996; 9 (01) 33-38
    CrossrefPubMedGoogle Scholar
  • 46 Keshari RS, Jyoti A, Dubey M. , et al. Cytokines induced neutrophil extracellular traps formation: implication for the inflammatory disease condition. PLoS One 2012; 7 (10) e48111
    CrossrefPubMedGoogle Scholar
  • 47 Lapponi MJ, Carestia A, Landoni VI. , et al. Regulation of neutrophil extracellular trap formation by anti-inflammatory drugs. J Pharmacol Exp Ther 2013; 345 (03) 430-437
    CrossrefPubMedGoogle Scholar
  • 48 Lau SY, Guild SJ, Barrett CJ. , et al. Tumor necrosis factor-alpha, interleukin-6, and interleukin-10 levels are altered in preeclampsia: a systematic review and meta-analysis. Am J Reprod Immunol 2013; 70 (05) 412-427
    PubMedGoogle Scholar
  • 49 Xu J, Lupu F, Esmon CT. Inflammation, innate immunity and blood coagulation. Hamostaseologie 2010; 30 (01) 5-6
    Article in Thieme ConnectPubMedGoogle Scholar
  • 50 Iwanaga S. The molecular basis of innate immunity in the horseshoe crab. Curr Opin Immunol 2002; 14 (01) 87-95
    CrossrefPubMedGoogle Scholar
  • 51 Fuchs TA, Bhandari AA, Wagner DD. Histones induce rapid and profound thrombocytopenia in mice. Blood 2011; 118 (13) 3708-3714
    CrossrefPubMedGoogle Scholar
  • 52 Kitzmiller JL, Benirschke K. Immunofluorescent study of placental bed vessels in pre-eclampsia of pregnancy. Am J Obstet Gynecol 1973; 115 (02) 248-251
    CrossrefPubMedGoogle Scholar
  • 53 Jiang R, Cai J, Zhu Z. , et al. Hypoxic trophoblast HMGB1 induces endothelial cell hyperpermeability via the TRL-4/caveolin-1 pathway. J Immunol 2014; 193 (10) 5000-5012
    CrossrefPubMedGoogle Scholar