PDF herunterladen
Thromb Haemost 2010; 104(02): 350-354
DOI: 10.1160/TH09-12-0816
Animal Models
Schattauer GmbH

A combined immunostimulatory and immunoinhibitory short interference RNA reduces hypercoagulability in a rat model of acute promyelocytic leukaemia

Per Ole Iversen
1   Department of Nutrition, Institute of Basic Medical Sciences, Faculty of Medicine, University of Oslo, Norway
2   Department of Hematology, Oslo University Hospital, Ullevål, Oslo, Norway
,
Dag Reidar Sørensen
3   Center for Comparative Medicine, Oslo University Hospital, Rikshospitalet, Oslo, Norway
,
Mouldy Sioud
4   Department of Immunology, Molecular Medicine Group, Oslo University Hospital, Radiumhospitalet, Oslo, Norway
› Institutsangaben Financial support: We appreciate the financial support from The Throne Holst Foundation.
Weitere Informationen

Publikationsverlauf

Received: 03. Dezember 2009

Accepted after major revision: 06. April 2010

Publikationsdatum:
24. November 2017 (online)

  als PDF herunterladen  Lizenzen und Reprints

Summary

Acute promyelocytic leukaemia (APL) confers an increased risk of thrombosis and bleeding. Current treatments are insufficient to inhibit these complications. We recently showed that a combined immunoinhibitory and immunostimulatory short interference (si) RNA effectively inhibited leukaemic growth and metastasis in rats with APL. We now asked if the reported anti-leukaemic effects of siRNA treatment could be explained by inhibition of hypercoagulability. We measured markers of coagulation and fibrinolysis in plasma collected from APL rats with overt leukaemia using conventional assays. Coagulopathy developed in untreated leukaemic rats evidenced by increase in several haemostatic markers. Treatment of leukaemic rats with the siRNA reduced (p < 0.05) the concentration of thrombin-anti-thrombin complex (a marker of coagulation) by 40% compared with rats treated with an inactive, control siRNA. Substantial reductions (p < 0.05) were also obtained for two markers of fibrinolysis: D-dimer (72%) and plasminogen activator inhibitor type 1 (51%). The activity of tissue factor, the main initiator of coagulation, was not increased (p > 0.05) in untreated leukaemic rats compared with healthy rats, and did not change (p > 0.05) upon treatment with the siRNA. The bifunctional siRNA reduces the hypercoagulable state in APL in addition to its direct anti-leukaemic properties, supporting testing of this small molecule in human APL.

Keywords

Acute myeloid leukaemia - coagulopathy - rat - short interference RNA
 

  • References

  • 1 Falanga A, Barbui T. Coagulopathy of acute promyelocytic leukemia. Acta Haematol 2001; 106: 43-51.
    CrossrefPubMedGoogle Scholar
  • 2 Stein E, McMahon B, Kwaan H. et al. The coagulopathy of acute promyelocytic leukaemia revisited. Best Pract Res Clin Haematol 2009; 22: 153-163.
    CrossrefPubMedGoogle Scholar
  • 3 Tallman MS, Brenner B, Serna JL. et al. Meeting report. Acute promyelocytic leukemia-associated coagulopathy, 21 January 2004, London, United Kingdom. Leuk Res 2005; 29: 347-351.
    CrossrefPubMedGoogle Scholar
  • 4 Falanga A, Rickles FR. Pathogenesis and management of the bleeding diathesis in acute promyelocytic leukaemia. Best Pract Res Clin Haematol 2003; 16: 463-482.
    CrossrefPubMedGoogle Scholar
  • 5 Falanga A, Consonni R, Marchetti M. et al. Cancer procoagulant and tissue factor are differently modulated by all-trans-retinoic acid in acute promyelocytic leukemia cells. Blood 1998; 92: 143-151.
    PubMedGoogle Scholar
  • 6 Falanga A, Barbui T, Rickles FR. Hypercoagulability and tissue factor gene upregulation in hematologic malignancies. Semin Thromb Hemost 2008; 34: 204-210.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 7 El-Shami K, Smith BD. Immunotherapy for myeloid leukemias: current status and future directions. Leukemia 2008; 22: 1658-1664.
    CrossrefPubMedGoogle Scholar
  • 8 Barrett AJ, Rezvani K. Translational mini-review series on vaccines: Peptide vaccines for myeloid leukaemias. Clin Exp Immunol 2007; 148: 189-198.
    CrossrefPubMedGoogle Scholar
  • 9 Neilson JR, Sharp PA. Small RNA regulators of gene expression. Cell 2008; 134: 899-902.
    CrossrefPubMedGoogle Scholar
  • 10 Furset G, Sioud M. Design of bifunctional siRNAs: combining immunostimulation and gene-silencing in one single siRNA molecule. Biochem Biophys Res Commun 2007; 352: 642-649.
    CrossrefPubMedGoogle Scholar
  • 11 Iversen PO, Semaeva E, Sorensen DR. et al. Dendritic cells loaded with tumor antigens and a dual immunostimulatory and anti-interleukin-10-specific small interference RNA prime T lymphocytes against leukemic cells. Transl Onc 2009; 02: 242-246.
    PubMedGoogle Scholar
  • 12 Sioud M. Induction of inflammatory cytokines and interferon responses by double-stranded and single-stranded siRNAs is sequence-dependent and requires endosomal localization. J Mol Biol 2005; 348: 1079-1090.
    CrossrefPubMedGoogle Scholar
  • 13 Sioud M, Furset G, Cekaite L. Suppression of immunostimulatory siRNA-driven innate immune activation by 2'-modified RNAs. Biochem Biophys Res Commun 2007; 361: 122-126.
    CrossrefPubMedGoogle Scholar
  • 14 Rehbinder C, Baneux P, Forbes D. et al. FELASA recommendations for the health monitoring of mouse, rat, hamster, gerbil, guinea pig and rabbit experimental units. Report of the Federation of European Laboratory Animal Science Associations (FELASA) Working Group on Animal Health accepted by the FELASA Board of Management, November 1995. Lab Anim 1996; 30: 193-208.
    CrossrefPubMedGoogle Scholar
  • 15 Lemini C, Jaimez R, Franco Y. Gender and interspecies influence on coagulation tests of rats and mice. Thromb Res 2007; 120: 415-419.
    CrossrefPubMedGoogle Scholar
  • 16 Lewis DA, Nyska A, Potti A. et al. Hemostatic activation in a chemically induced rat model of severe hemolysis and thrombosis. Thromb Res 2006; 118: 747-753.
    CrossrefPubMedGoogle Scholar
  • 17 van Westerloo DJ, Giebelen IA, Meijers JC. et al. Vagus nerve stimulation inhibits activation of coagulation and fibrinolysis during endotoxemia in rats. J Thromb Haemost 2006; 04: 1997-2002.
    CrossrefPubMedGoogle Scholar
  • 18 Surprenant YM, Zuckerman SH. A novel microtiter plate assay for the quantitation of procoagulant activity on adherent monocytes, macrophage and endothelial cells. Thromb Res 1989; 53: 339-346.
    CrossrefPubMedGoogle Scholar
  • 19 Kochetkova M, Iversen PO, Lopez AF, Shannon MF. Deoxyribonucleic acid triplex formation inhibits granulocyte macrophage colony-stimulating factor gene expression and suppresses growth in juvenile myelomonocytic leukemic cells. J Clin Invest 1997; 99: 3000-3008.
    CrossrefPubMedGoogle Scholar
  • 20 Martens AC, van Bekkum DW, Hagenbeek A. The BN acute myelocytic leukemia (BNML) (a rat model for studying human acute myelocytic leukemia (AML)). Leukemia 1990; 04: 241-257.
    PubMedGoogle Scholar
  • 21 Mc CE, Bruserud O, Gjertsen BT. Animal models of acute myelogenous leukaemia – development, application and future perspectives. Leukemia 2005; 19: 687-706.
    CrossrefPubMedGoogle Scholar
  • 22 Falanga A, Panova-Noeva M, Russo L. Procoagulant mechanisms in tumour cells. Best Pract Res Clin Haematol 2009; 22: 49-60.
    CrossrefPubMedGoogle Scholar
  • 23 Colucci M, Lorenzet R, Locati D. et al. Occurrence of disseminated intravascular coagulation in rat BNML leukaemia despite lack of leucocyte procoagulant activity. Br J Exp Pathol 1983; 64: 207-210.
    PubMedGoogle Scholar
  • 24 Negaard HF, Iversen PO, Ostenstad B. et al. Hypercoagulability in patients with haematological neoplasia: no apparent initiation by tissue factor. Thromb Hae-most 2008; 99: 1040-1048.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 25 Foss B, Bruserud O. Platelet functions and clinical effects in acute myelogenous leukemia. Thromb Haemost 2008; 99: 27-37.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 26 Soslau G, Morgan DA, Jaffe JS. et al. Cytokine mRNA expression in human platelets and a megakaryocytic cell line and cytokine modulation of platelet function. Cytokine 1997; 09: 405-411.
    CrossrefPubMedGoogle Scholar