Advertisement

ENMD-1068, a protease-activated receptor 2 antagonist, inhibits the development of endometriosis in a mouse model

Published:February 03, 2014DOI:https://doi.org/10.1016/j.ajog.2014.01.040

      Objective

      Protease-activated receptor 2 plays an important role in the pathogenesis of endometriosis. We studied the effect of ENMD-1068, a protease-activated receptor 2 antagonist, on the development of endometriosis in a noninvasive fluorescent mouse model.

      Study Design

      A red fluorescent protein–expressing xenograft model of human endometriosis was created in nude mice. After endometriosis induction, the mice were injected intraperitoneally with either 25 mg/kg or 50 mg/kg ENMD-1068 or with 200 μL of the vehicle control daily for 5 days. The endometriotic lesions that developed in the mice were then counted, measured, and collected. The lesions were assessed for the production of interleukin 6 and monocyte chemotactic protein-1 by enzyme-linked immunosorbent assays and evaluated for the activation of nuclear factor-κB and the expression of vascular endothelial growth factor by immunohistochemical analyses. Cell proliferation and apoptosis were assessed by immunohistochemistry for Ki-67 and terminal deoxynucleotidyl transferase–mediated dUTP nick end labeling, respectively.

      Results

      ENMD-1068 dose-dependently inhibited the development of endometriotic lesions (P < .05) without apparent toxicity to various organs of the treated mice. Consistently, ENMD-1068 dose-dependently inhibited the expression of interleukin 6 and nuclear factor-κB (P < .05) and cell proliferation (P < .05) in the lesions, as well as increased the percentage of apoptotic cells (P < .05). ENMD-1068 reduced the levels of monocyte chemotactic protein-1 and vascular endothelial growth factor in the lesions (P < .05), but not in a dose-dependent manner.

      Conclusion

      Our study suggests that ENMD-1068 is effective in suppressing the growth of endometriosis, which might be attributed to the drug's antiangiogenic and antiinflammatory activities.

      Key words

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to American Journal of Obstetrics & Gynecology
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Bulun S.E.
        Endometriosis.
        N Engl J Med. 2009; 360: 268-279
        • Munksgaard P.S.
        • Blaakaer J.
        The association between endometriosis and ovarian cancer: a review of histological, genetic and molecular alterations.
        Gynecol Oncol. 2012; 124: 164-169
        • Kumar S.
        • Munkarah A.
        • Arabi H.
        • et al.
        Prognostic analysis of ovarian cancer associated with endometriosis.
        Am J Obstet Gynecol. 2011; 204: 63.e1-63.e7
        • Hansen K.A.
        • Eyster K.M.
        Genetics and genomics of endometriosis.
        Clin Obstet Gynecol. 2010; 53: 403-412
        • Camerer E.
        • Trejo J.
        Cryptic messages: is noncoagulant tissue factor reserved for cell signaling?.
        Proc Natl Acad Sci U S A. 2006; 103: 14259-14260
        • Hirota Y.
        • Osuga Y.
        • Hirata T.
        • et al.
        Activation of protease- activated receptor 2 stimulates proliferation and interleukin (IL)-6 and IL-8 secretion of endometriotic stromal cells.
        Hum Reprod. 2005; 20: 3547-3553
        • Osuga Y.
        • Hirota Y.
        • Yoshino O.
        • Hirata T.
        • Koga K.
        • Taketani Y.
        Proteinase-activated receptors in the endometrium and endometriosis.
        Front Biosci. 2012; 4: 755-767
        • Osuga Y.
        • Hirota Y.
        • Taketani Y.
        Basic and translational research on proteinase-activated receptors: proteinase-activated receptors in female reproductive tissues and endometriosis.
        J Pharmacol Sci. 2008; 108: 422-425
        • Steinhoff M.
        • Vergnolle N.
        • Young S.H.
        • et al.
        Agonists of proteinase-activated receptor 2 induce inflammation by a neurogenic mechanism.
        Nat Med. 2000; 6: 151-158
        • Crilly A.
        • Palmer H.
        • Nickdel M.B.
        • et al.
        Immunomodulatory role of proteinase-activated receptor-2.
        Ann Rheum Dis. 2012; 71: 1559-1566
        • Lin M.
        • Weng H.
        • Wang X.
        • Zhou B.
        • Yu P.
        • Wang Y.
        The role of tissue factor and protease-activated receptor 2 in endometriosis.
        Am J Reprod Immunol. 2012; 68: 251-257
        • Krikun G.
        • Hu Z.
        • Osteen K.
        • et al.
        The immunoconjugate “icon” targets aberrantly expressed endothelial tissue factor causing regression of endometriosis.
        Am J Pathol. 2010; 176: 1050-1056
        • Kelso E.B.
        • Lockhart J.C.
        • Hembrough T.
        • et al.
        Therapeutic promise of proteinase-activated receptor-2 antagonism in joint inflammation.
        J Pharmacol Exp Ther. 2006; 316: 1017-1024
        • Kaufmann R.
        • Mussbach F.
        • Henklein P.
        • Settmacher U.
        Proteinase-activated receptor 2 mediated calcium signaling in hepatocellular carcinoma cells.
        J Cancer Res Clin Oncol. 2011; 137: 965-973
        • Grümmer R.
        • Schwarzer F.
        • Bainczyk K.
        • et al.
        Peritoneal endometriosis: validation of an in-vivo model.
        Hum Reprod. 2001; 16: 1736-1743
        • Fortin M.
        • Lépine M.
        • Pagé M.
        • et al.
        An improved mouse model for endometriosis allows noninvasive assessment of lesion implantation and development.
        Fertil Steril. 2003; 80: 832-838
        • Gao X.
        • Chen J.
        • Chen J.
        • Wu B.
        • Chen H.
        • Jiang X.
        Quantum dots bearing lectin-functionalized nanoparticles as a platform for in vivo brain imaging.
        Bioconjug Chem. 2008; 19: 2189-2195
        • Carlsson G.
        • Gullberg B.
        • Hafström L.
        Estimation of liver tumor volume using different formulas–an experimental study in rats.
        J Cancer Res Clin Oncol. 1983; 105: 20-23
        • Komiyama S.
        • Aoki D.
        • Katsuki Y.
        • Nozawa S.
        Proliferative activity of early ovarian clear cell adenocarcinoma depends on association with endometriosis.
        Eur J Obstet Gynecol Reprod Biol. 2006; 127: 130-136
        • Grümmer R.
        Animal models in endometriosis research.
        Hum Reprod Update. 2006; 12: 641-649
        • Sampson J.A.
        Metastatic or embolic endometriosis, due to the menstrual dissemination of endometrial tissue into the venous circulation.
        Am J Pathol. 1927; 3: 93-110
        • Bulletti C.
        • DeZiegler D.
        • Stefanetti M.
        • Cicinelli E.
        • Pelosi E.
        • Flamigni C.
        Endometriosis: absence of recurrence in patients after endometrial ablation.
        Hum Reprod. 2001; 16: 2676-2679
        • Liu B.
        • Wang N.N.
        • Wang Z.L.
        • et al.
        Improved nude mouse models for green fluorescence human endometriosis.
        J Obstet Gynaecol Res. 2010; 36: 1214-1221
        • Altan Z.M.
        • Denis D.
        • Kagan D.
        • Grund E.M.
        • Palmer S.S.
        • Nataraja S.G.
        A long-acting tumor necrosis factor alpha-binding protein demonstrates activity in both in vitro and in vivo models of endometriosis.
        J Pharmacol Exp Ther. 2010; 334: 460-466
        • Stuelten C.H.
        • Kamaraju A.K.
        • Wakefield L.M.
        • Roberts A.B.
        Lentiviral reporter constructs for fluorescence tracking of the temporospatial pattern of Smad3 signaling.
        Biotechniques. 2007; 43 (292, 294): 289-290
        • Shinohara A.
        • Kutsukake M.
        • Takahashi M.
        • Kyo S.
        • Tachikawa E.
        • Tamura K.
        Protease-activated receptor-stimulated interleukin-6 expression in endometriosis-like lesions in an experimental mouse model of endometriosis.
        J Pharmacol Sci. 2012; 119: 40-51
        • Othman Ewl-D.
        • Hornung D.
        • Aalem H.T.
        • Khalifa E.A.
        • El-Metwally T.H.
        • Al-Hendy A.
        Serum cytokines as biomarkers for nonsurgical prediction of endometriosis.
        Eur J Obstet Gynecol Reprod Biol. 2008; 137: 240-246
        • Elgafor E.I.
        • Sharkwy I.A.
        Combination of non-invasive and semi-invasive tests for diagnosis of minimal to mild endometriosis.
        Arch Gynecol Obstet. 2013; 288: 793-797
        • Sidell N.
        • Han S.W.
        • Parthasarathy S.
        Regulation and modulation of abnormal immune responses in endometriosis.
        Ann N Y Acad Sci. 2002; 955: 159-173
        • Gu L.
        • Tseng S.
        • Horner R.M.
        • Tam C.
        • Loda M.
        • Rollins B.J.
        Control of TH2 polarization by the chemokine monocyte chemoattractant protein-1.
        Nature. 2000; 404: 407-411
        • Selam B.
        • Kayisli U.A.
        • Akbas G.E.
        • Basar M.
        • Arici A.
        Regulation of FAS ligand expression by chemokine ligand 2 in human endometrial cells.
        Biol Reprod. 2006; 75: 203-209
        • Lin Y.J.
        • Lai M.D.
        • Lei H.Y.
        • Wing L.Y.
        Neutrophils and macrophages promote angiogenesis in the early stage of endometriosis in a mouse model.
        Endocrinology. 2006; 147: 1278-1286
        • Lin J.
        • Gu Y.
        Effect of monocyte chemoattractant protein-1 and estradiol on the secretion of vascular endothelial growth factor in endometrial stromal cells in vitro.
        Fertil Steril. 2005; 84: 1793-1796
        • Agic A.
        • Xu H.
        • Finas D.
        • Banz C.
        • Diedrich K.
        • Hornung D.
        Is endometriosis associated with systemic subclinical inflammation?.
        Gynecol Obstet Invest. 2006; 62: 139-147
        • Fasciani A.
        • D'Ambrogio G.
        • Bocci G.
        • Monti M.
        • Genazzani A.R.
        • Artini P.G.
        High concentration of the vascular endothelial growth factor and interleukin-8 in ovarian endometrioma.
        Mol Hum Reprod. 2000; 6: 50-54
        • Tennant G.M.
        • Wadsworth R.M.
        • Kennedy S.
        PAR-2 mediates increased inflammatory cell adhesion and neointima formation following vascular injury in the mouse.
        Atherosclerosis. 2008; 198: 57-64
        • Ruf W.
        • Disse J.
        • Carneiro-Lobo T.C.
        • Yokota N.
        • Schaffner F.
        Tissue factor and cell signaling in cancer progression and thrombosis.
        J Thromb Haemost. 2011; 9: 306-315
        • González-Ramos R.
        • Van Langendonckt A.
        • Defrère S.
        • et al.
        Involvement of the nuclear factor-κB pathway in the pathogenesis of endometriosis.
        Fertil Steril. 2010; 94: 1985-1994
        • Kiriakidis S.
        • Andreakos E.
        • Monaco C.
        • Foxwell B.
        • Feldmann M.
        • Paleolog E.
        VEGF expression in human macrophages is NF-κB dependent: studies using adenoviruses expressing the endogenous NF-κB inhibitor IκBa and a kinase defective form of the IκB kinase 2.
        J Cell Sci. 2003; 116: 665-674
        • Guo D.
        • Zhou H.
        • Wu Y.
        • et al.
        Involvement of ERK1/2/NF-κB signal transduction pathway in TF/FVIIa/PAR2-induced proliferation and migration of colon cancer cell SW620.
        Tumour Biol. 2011; 32: 921-930
        • González-Ramos R.
        • Van Langendonckt A.
        • Defrère S.
        • et al.
        Agents blocking the nuclear factor-kappaB pathway are effective inhibitors of endometriosis in an in vivo experimental model.
        Gynecol Obstet Invest. 2008; 65: 174-186