NF-κB Decoy Oligonucleotides Suppress RANTES Expression and Monocyte Chemotactic Activity via NF-κB Inactivation in Stromal Cells of Ectopic Endometrium

Background The nuclear factor kappa B (NF-κB) pathway is a critical mediator of regulated on activation, normal T cell expressed and secreted (RANTES) gene regulation and therefore represents a potential target for therapy of endometriosis-associated symptoms.

The objective of this study was to investigate the effect of NF-κB decoy oligonucleotides (ODNs) on NF-κB activation, RANTES expression, and monocyte chemotactic activity in ectopic endometrial stromal cells in vitro.

A specific sandwich enzyme-linked immunosorbent assay (ELISA) was used to quantify RANTES expression in ectopic and normal endometrial stromal cells stimulated by interleukin (IL)-1β. Four hours after transfection of NF-κB decoy ODNs, 10 ng/ml IL-1β was added to induce the ectopic endometrial stromal cells to secrete RANTES. The NF-κB activation, RANTES expression, and monocyte chemotactic activity in ectopic endometrial stromal cells were respectively evaluated by electrophoretic mobility shift assay, ELISA, and Boyden chambers.

IL-1β induced significantly higher levels (P < 0.05) of RANTES expression in a time-dependent manner in ectopic endometrial stromal cells compared with IL-1β-untreated ectopic and normal endometrial stromal cells. The RANTES accounts for the majority (68%) of the monocyte chemotactic activity in conditioned media of ectopic endometrial stromal cells. In vitro transfection of NF-κB decoy ODNs dramatically decreased (P < 0.05) the NF-κB activation, RANTES expression, and monocyte chemotactic activity in IL-1β-induced ectopic endometrial stromal cells.

NF-κB decoy ODNs may exert anti-inflammatory effects in ectopic endometrial stromal cells via the suppression of NF-κB activation, RANTES expression, and monocyte chemotactic activity. Similar content being viewed by others

Herrmann Lavoie C, Fraser D, Therriault MJ, Akoum A. Interleukin-1 stimulates macrophage migration inhibitory factor secretion in ectopic endometrial cells of women with endometriosis. Am J Reprod Immunol 2007;58(6):505–13. doi:10.1111/j.1600-0897.2007.00471.x. Lebovic DI, Mueller MD, Taylor RN. Immunobiology of endometriosis. Fertil Steril 2001;75(1):1–10. doi:10.1016/S0015-0282(00)01630-7. Na YJ, Jin JO, Lee MS, Song MG, Lee KS, Kwak JY. Peritoneal fluid from endometriosis patients switches differentiation of monocytes from dendritic cells to macrophages. J Reprod Immunol 2008;77(1):63–74. doi:10.1016/j.jri.2007.03.013. Bersinger NA, von Roten S, Wunder DM, Raio L, Dreher E, Mueller MD. PAPP-A and osteoprotegerin, together with interleukin-8 and RANTES, are elevated in the peritoneal fluid of women with endometriosis. Am J Obstet Gynecol 2006;195(1):103–8. doi:10.1016/j.ajog.2005.12.010. Khorram O, Taylor RN, Ryan IP, Schall TJ, Landers DV. Peritoneal fluid concentrations of the cytokine RANTES correlate with the severity of endometriosis. Am J Obstet Gynecol 1993;169:1545–9. Hornung D, Bentzien F, Wallwiener D, Kiesel L, Taylor RN. Chemokine bioactivity of RANTES in endometriotic and normal endometrial stromal cells and peritoneal fluid. Mol Hum Reprod 2001;7:163–8. doi:10.1093/molehr/7.2.163. Lebovic DI, Chao VA, Taylor RN. Peritoneal macrophages induce RANTES (regulated on activation, normal T cell expressed and secreted) chemokine gene transcription in endometrial stromal cells. J Clin Endocrinol Metab 2004;89(3):1397–401. doi:10.1210/jc.2003-031010. Hornung D, Klingel K, Dohrn K, Kandolf R, Wallwiener D, Taylor RN. Regulated on activation, normal T-cell-expressed and -secreted mRNA expression in normal endometrium and endometriotic implants: assessment of autocrine/paracrine regulation by in situ hybridization. Am J Pathol 2001;158(6):1949–54. Hornung D, Chao VA, Vigne JL, Wallwiener D, Taylor RN. Thiazolidinedione inhibition of peritoneal inflammation. Gynecol Obstet Invest 2003;55:20–4. doi:10.1159/000068952. Lebovic DI, Chao VA, Martini JF, Taylor RN. IL-1beta induction of RANTES (regulated upon activation, normal T cell expressed and secreted) chemokine gene expression in endometriotic stromal cells depends on a nuclear factor-kappa B site in the proximal promoter. J Clin Endocrinol Metab 2001;86(10):4759–64. doi:10.1210/jc.86.10.4759. King AE, Critchley HO, Kelly RW. The NF-κB pathway in human endometrium and first trimester decidua. Mol Hum Reprod 2001;7:175–83. doi:10.1093/molehr/7.2.175. Laird SM, Tuckerman EM, Cork BA, Li TC. Expression of nuclear factor κB in human endometrium; role in the control of interleukin 6 and leukaemia inhibitory factor production. Mol Hum Reprod 2000;6:34–40. doi:10.1093/molehr/6.1.34. American Society for Reproductive Medicine. Revised American Society for Reproductive Medicine classification of endometriosis: 1996. Fertil Steril 1997;67:817–21. doi:10.1016/S0015-0282(97)81391-X. Ryan IP, Schriock ED, Taylor RN. Isolation, characterization, and comparison of human endometrial and endometriosis cells in vitro. J Clin Endocrinol Metab 1994;78(3):642–9. doi:10.1210/jc.78.3.642. Morishita R, Sugimoto T, Aoki M, Kita I, Tomita N, Moriguchi A, Maeda K, Sawa Y, Kaneda Y, Higaki J, Ogihara T. In vivo transfection of cis element “decoy” against nuclear factor-κB binding site prevents myocardial infarction. Nat Med 1997;3:894–9. doi:10.1038/nm0897-894. Kunugiza Y, Tomita T, Tomita N, Morishita R, Yoshikawa H. Inhibitory effect of ribbon-type NF-kappa B decoy oligodeoxynucleotides on osteoclast induction and activity in vitro and in vivo. Arthritis Res Ther 2006;8(4):R103. Akoum A, Lawson C, Herrmann-Lavoie C, Maheux R. Imbalance in the expression of the activating type I and the inhibitory type II interleukin 1 receptors in endometriosis. Hum Reprod 2007;22(5):1464–73. doi:10.1093/humrep/dem021. Stoffel A. The NF-kappa B signalling pathway: a therapeutic target in lymphoid malignancies? Expert Opin Ther Targets 2005;9(5):1045–61. doi:10.1517/14728222.9.5.1045. González-Ramos R, Donnez J, Defrère S, Leclercq I, Squifflet J, Lousse JC, Van Langendonckt A. Nuclear factor-kappa B is constitutively activated in peritoneal endometriosis. Mol Hum Reprod 2007;13(7):503–9. doi:10.1093/molehr/gam033. Lousse JC, Van Langendonckt A, González-Ramos R, Defrère S, Renkin E, Donnez J. Increased activation of nuclear factor-kappa B (NF-kappa B) in isolated peritoneal macrophages of patients with endometriosis. Fertil Steril 2008;90(1):217–20. doi:10.1016/j.fertnstert.2007.06.015. Lindström TM, Bennett PR. The role of nuclear factor kappa B in human labour. Reproduction 2005;130(5):569–81. doi:10.1530/rep.1.00197. Grund EM, Kagan D, Tran CA, Zeitvogel A, Starzinski-Powitz A, Nataraja S, Palmer SS. Tumor necrosis factor-alpha regulates inflammatory and mesenchymal responses via mitogen-activated protein kinase kinase, p38, and nuclear factor kappa B in human endometriotic epithelial cells. Mol Pharmacol 2008;73(5):1394–404. doi:10.1124/mol.107.042176. Sakamoto Y, Harada T, Horie S, Iba Y, Taniguchi F, Yoshida S, Iwabe T, Terakawa N. Tumor necrosis factor-alpha-induced interleukin-8 (IL-8) expression in endometriotic stromal cells, probably through nuclear factor-kappa B activation: gonadotropin-releasing hormone agonist treatment reduced IL-8 expression. Clin Endocrinol Metab 2003;88(2):730–5. doi:10.1210/jc.2002-020666. Yamauchi N, Harada T, Taniguchi F, Yoshida S, Iwabe T, Terakawa N. Tumor necrosis factor-alpha induced the release of interleukin-6 from endometriotic stromal cells by the nuclear factor-kappa B and mitogen-activated protein kinase pathways. Fertil Steril 2004;82(Suppl 3):1023–8. doi:10.1016/j.fertnstert.2004.02.134. Horie S, Harada T, Mitsunari M, Taniguchi F, Iwabe T, Terakawa N. Progesterone and progestational compounds attenuate tumor necrosis factor alpha-induced interleukin-8 production via nuclear factor kappa B inactivation in endometriotic stromal cells. Fertil Steril 2005;83(5):1530–5. doi:10.1016/j.fertnstert.2004.11.042. Yagyu T, Kobayashi H, Matsuzaki H, Wakahara K, Kondo T, Kurita N, Sekino H, Inagaki K, Suzuki M, Kanayama N, Terao T. Thalidomide inhibits tumor necrosis factor-alpha-induced interleukin-8 expression in endometriotic stromal cells, possibly through suppression of nuclear factor-kappa B activation. J Clin Endocrinol Metab 2005;90(5):3017–21. doi::10.1210/jc.2004-1946. Wieser F, Vigne JL, Ryan I, Hornung D, Djalali S, Taylor RN. Sulindac suppresses nuclear factor-kappa B activation and RANTES gene and protein expression in endometrial stromal cells from women with endometriosis. J Clin Endocrinol Metab 2005;90(12):6441–7. doi:10.1210/jc.2005-0972. Nasu K, Nishida M, Ueda T, Yuge A, Takai N, Narahara H. Application of the nuclear factor-kappa B inhibitor BAY 11-7085 for the treatment of endometriosis: an in vitro study. Am J Physiol Endocrinol Metab 2007;293(1):E16–23. doi:10.1152/ajpendo.00135.2006. Celik O, Hascalik S, Elter K, Tagluk ME, Gurates B, Aydin NE. Combating endometriosis by blocking proteasome and nuclear factor-{kappa}B pathways. Hum Reprod 2008;23:2458–65, Aug 2. González-Ramos R, Van Langendonckt A, Defrère S, Lousse JC, Mettlen M, Guillet A, Donnez J. Agents blocking the nuclear factor-kappa B pathway are effective inhibitors of endometriosis in an in vivo experimental model. Gynecol Obstet Invest 2008;65(3):174–86. doi:10.1159/000111148. Bielinska A, Shivdasani RA, Zhang LQ, Nabel GJ. Regulation of gene expression with double-stranded phosphorothioate oligonucleotides. Science 1990;250(4983):997–1000. doi:10.1126/science.2237444. Mann MJ, Dzau VJ. Therapeutic applications of transcription factor decoy oligonucleotides. J Clin Invest 2000;106(9):1071–5. doi:10.1172/JCI11459. Matsuda N, Hattori Y, Jesmin S, Gando S. Nuclear factor-kappa B decoy oligodeoxynucleotides prevent acute lung injury in mice with cecal ligation and puncture-induced sepsis. Mol Pharmacol 2005;67(4):1018–25. doi:10.1124/mol.104.005926. Nakagami H, Tomita N, Kaneda Y, Ogihara T, Morishita R. Anti-oxidant gene therapy by NF kappa B decoy oligodeoxynucleotide. Curr Pharm Biotechnol 2006;7(2):95–100. doi:10.2174/138920106776597702. Penolazzi L, Magri E, Lambertini E, Calò G, Cozzani M, Siciliani G, Piva R, Gambari R. Local in vivo administration of a decoy oligonucleotide targeting NF-kappa B induces apoptosis of osteoclasts after application of orthodontic forces to rat teeth. Int J Mol Med 2006;18(5):807–11. Tomita N, Azuma H, Kaneda Y, Ogihara T, Morishita R. Application of decoy oligodeoxynucleotides-based approach to renal diseases. Curr Drug Targets 2004;5(8):717–33. doi:10.2174/1389450043345146. Xu MQ, Shuai XR, Yan ML, Zhang MM, Yan LN. Nuclear factor-kappa B decoy oligodeoxynucleotides attenuates ischemia/reperfusion injury in rat liver graft. World J Gastroenterol 2005;11(44):6960–7. Acknowledgments We thank Dr. Lu Hua for the help in gel mobility shift assay. Funding This work is supported by the National Natural Science Foundation of China No 30600671 (to Wang Xiu-li) and the Natural Science Foundation of Jiangsu province no BK2006577 (to Wang Xiu-li). Author information Authors and Affiliations Corresponding author Rights and permissions About this article Cite this article Xiu-li, W., Su-ping, H., Hui-hua, D. et al. NF-κB Decoy Oligonucleotides Suppress RANTES Expression and Monocyte Chemotactic Activity via NF-κB Inactivation in Stromal Cells of Ectopic Endometrium. J Clin Immunol 29, 387–395 (2009). https://doi.org/10.1007/s10875-009-9274-z Received: Accepted: Published: Issue date: DOI: https://doi.org/10.1007/s10875-009-9274-z