Endometriosis is caused by the displacement of endometrium outside the uterus contributing heavily to infertility and debilitating pelvic pain. Ectopic adhesion and growth are believed to occur under the influence of a favorable hormonal environment and immunological factors. The objective of this study is to analyze the effect of a targeted therapy with an antibody-based pharmacodelivery of interleukin 4 (F8-IL4) in a mouse model of experimentally induced endometriosis. Endometriosis-like lesions were induced in Balb/c mice. The animals were treated intravenously with F8-IL4 or with untargeted IL4 (KSF-IL4). Twelve days after disease induction, the lesions were isolated. A significant reduction in the number of total lesions/mouse and in the total volume of lesions/mouse was observed in mice treated with F8-IL4 compared to controls (P = .029 and P = .006, respectively), while no difference was found between KSF-IL4-treated mice and their controls. Gene expression was evaluated by quantitative real-time polymerase chain reaction. Expression of genes involved in cell adhesion, extracellular matrix invasion, and neovascularization was significantly downregulated in F8-IL4-treated mice compared to their controls (integrin β1: P = .02; metalloproteinase [MMP] 3: P = .02; MMP9: P = .04; vascular endothelial growth factor: P = .04). Gene expression of inflammatory cytokines (tumor necrosis factor α, IL1β, IL1α, and IL6) did not vary in the ectopic lesions isolated from F8-IL4-treated mice compared to their controls. Immunohistochemistry demonstrated a significantly reduced expression of E-cadherin and β-catenin in the lesions of mice treated with F8-IL4. Our results show that the antibody-mediated targeted delivery of IL4 inhibits the development of endometriosis in a syngeneic mouse model by likely impairing adhesion, invasion, and vascularization of the ectopic endometrium.
MeulemanCVandenabeeleBFieuwsSSpiessensCTimmermanDD’HoogheT. High prevalence of endometriosis in infertile women with normal ovulation and normospermic partners. Fertil Steril. 2009;92(1):68–74.
2.
BulunSE. Endometriosis. N Engl J Med. 2009;360(3):268–279.
3.
BurneyROGiudiceLC. Pathogenesis and pathophysiology of endometriosis. Fertil Steril. 2012;98(3):511–519.
4.
TsengJFRyanIPMilamTD. Interleukin-6 secretion in vitro is up-regulated in ectopic and eutopic endometrial stromal cells from women with endometriosis. J Clin Endocrinol Metab. 1996;81(3):1118–1122.
5.
WuYKajdacsy-BallaAStrawnE. Transcriptional characterizations of differences between eutopic and ectopic endometrium. Endocrinology. 2006;147(1):232–246.
6.
Sharpe-TimmsKLCoxKE. Paracrine regulation of matrix metalloproteinase expression in endometriosis. Ann N Y Acad Sci. 2002;955:147–156; discussion157–148, 396–406.
7.
AkoumAJolicoeurCBoucherA. Estradiol amplifies interleukin-1-induced monocyte chemotactic protein-1 expression by ectopic endometrial cells of women with endometriosis. J Clin Endocrinol Metab. 2000;85(2):896–904.
8.
NairASNairHBLucidiRS. Modeling the early endometriotic lesion: mesothelium-endometrial cell co-culture increases endometrial invasion and alters mesothelial and endometrial gene transcription. Fertil Steril. 2008;90(4 suppl):1487–1495.
9.
MayKEVillarJKirtleySKennedySHBeckerCM. Endometrial alterations in endometriosis: a systematic review of putative biomarkers. Hum Reprod Update. 2011;17(5):637–653.
10.
EdwardsAKNakamuraDSViraniSWesselsJMTayadeC. Animal models for anti-angiogenic therapy in endometriosis. J Reprod Immunol. 2013;97(1):85–94.
11.
LebovicDIMuellerMDTaylorRN. Immunobiology of endometriosis. Fertil Steril. 2001;75(1):1–10.
12.
BacciMCapobiancoAMonnoA. Macrophages are alternatively activated in patients with endometriosis and required for growth and vascularization of lesions in a mouse model of disease. Am J Pathol. 2009;175(2):547–556.
13.
CapobiancoARovere-QueriniP. Endometriosis, a disease of the macrophage. Front Immunol. 2013;4:9.
14.
MarianiMViganoPGentiliniD. The selective vitamin D receptor agonist, elocalcitol, reduces endometriosis development in a mouse model by inhibiting peritoneal inflammation. Hum Reprod. 2012;27(7):2010–2019.
15.
SomiglianaEViganoPRossiGCarinelliSVignaliMPanina-BordignonP. Endometrial ability to implant in ectopic sites can be prevented by interleukin-12 in a murine model of endometriosis. Hum Reprod. 1999;14(12):2944–2950.
16.
VercelliniPViganoPSomiglianaEFedeleL. Endometriosis: pathogenesis and treatment. Nature reviews. Endocrinology. 2014;10(5):261–275.
17.
GuoSW. An overview of the current status of clinical trials on endometriosis: issues and concerns. Fertil Steril. 2014;101(1):183–190e184.
18.
SchwagerKBootzFImeschPKasparMTrachselENeriD. The antibody-mediated targeted delivery of interleukin-10 inhibits endometriosis in a syngeneic mouse model. Hum Reprod. 2011;26(9):2344–2352.
19.
VillaATrachselEKasparM. A high-affinity human monoclonal antibody specific to the alternatively spliced EDA domain of fibronectin efficiently targets tumor neo-vasculature in vivo. Int J Cancer. 2008;122(11):2405–2413.
20.
SchwagerKKasparMBootzF. Preclinical characterization of DEKAVIL (F8-IL10), a novel clinical-stage immunocytokine which inhibits the progression of collagen-induced arthritis. Arthritis Res Ther. 2009;11(5):R142.
21.
RybakJNRoesliCKasparMVillaANeriD. The extra-domain A of fibronectin is a vascular marker of solid tumors and metastases. Cancer Res. 2007;67(22):10948–10957.
22.
SchliemannCWiedmerAPedrettiMSzczepanowskiMKlapperWNeriD. Three clinical-stage tumor targeting antibodies reveal differential expression of oncofetal fibronectin and tenascin-C isoforms in human lymphoma. Leukemia Res. 2009;33(12):1718–1722.
23.
GutbrodtKLSchliemannCGiovannoniL. Antibody-based delivery of interleukin-2 to neovasculature has potent activity against acute myeloid leukemia. Sci Transl Med. 2013;5(201):201ra118.
24.
FranzMHilgerIGrunK. Selective imaging of chronic cardiac rejection using a human antibody specific to the alternatively spliced EDA domain of fibronectin. J Heart Lung Transplant. 2013;32(6):641–650.
25.
FiechterMFreyKFugmannTKaufmannPANeriD. Comparative in vivo analysis of the atherosclerotic plaque targeting properties of eight human monoclonal antibodies. Atherosclerosis. 2011;214(2):325–330.
26.
HemmerleTZgraggenSMatasciMHalinCDetmarMNeriD. Antibody-mediated delivery of interleukin 4 to the neo-vasculature reduces chronic skin inflammation. J Dermatol Sci. 2014;76(2):96–103.
27.
PaulWE. What determines Th2 differentiation, in vitro and in vivo?Immunol Cell Biol. 2010;88(3):236–239.
28.
HemmerleTDollFNeriD. Antibody-based delivery of IL4 to the neovasculature cures mice with arthritis. Proc Natl Acad Sci U S A. 2014;111(33):12008–12012.
29.
HemmerleTNeriD. The antibody-based targeted delivery of interleukin-4 and 12 to the tumor neovasculature eradicates tumors in three mouse models of cancer. Int J Cancer. 2014;134(2):467–477.
30.
RicciAGOlivaresCNBilotasMAMeresmanGFBaranaoRI. Effect of vascular endothelial growth factor inhibition on endometrial implant development in a murine model of endometriosis. Reprod Sci. 2011;18(7):614–622.
31.
BesteMTPfaffle-DoyleNPrenticeEA. Molecular network analysis of endometriosis reveals a role for c-Jun-regulated macrophage activation. Sci Transl Med. 2014;6(222):222ra216.
32.
EfstathiouJASampsonDALevineZ. Nonsteroidal antiinflammatory drugs differentially suppress endometriosis in a murine model. Fertil Steril. 2005;83(1):171–181.
33.
MantovaniASicaALocatiM. New vistas on macrophage differentiation and activation. Eur J Immunol. 2007;37(1):14–16.
34.
De PalmaMNaldiniL. Tie2-expressing monocytes (TEMs): novel targets and vehicles of anticancer therapy?Biochimica et biophysica acta. 2009;1796(1):5–10.
35.
CapobiancoAMonnoACottoneL. Proangiogenic Tie2(+) macrophages infiltrate human and murine endometriotic lesions and dictate their growth in a mouse model of the disease. Am J Pathol. 2011;179(5):2651–2659.
36.
GarciaSKrauszSAmbarusCA. Tie2 signaling cooperates with TNF to promote the pro-inflammatory activation of human macrophages independently of macrophage functional phenotype. PloS One. 2014;9(1):e82088.
37.
DunselmanGAGroothuisPGde GoeijAFEversJL. The Mesothelium, Teflon or Velcro? Mesothelium in endometriosis pathogenesis. Hum Reprod. 2001;16(4):605–607.
38.
YoungVJBrownJKSaundersPTHorneAW. The role of the peritoneum in the pathogenesis of endometriosis. Hum Reprod Update. 2013;19(5):558–569.
39.
van der LindenPJde GoeijAFDunselmanGAvan der LindenEPRamaekersFCEversJL. Expression of integrins and E-cadherin in cells from menstrual effluent, endometrium, peritoneal fluid, peritoneum, and endometriosis. Fertil Steril. 1994;61(1):85–90.
40.
HullMLEscarenoCRGodslandJM. Endometrial-peritoneal interactions during endometriotic lesion establishment. Am J Pathol. 2008;173(3):700–715.
41.
HerzbergFSchoningMSchirnerMToppMThielEKreuserED. IL-4 and TNF-alpha induce changes in integrin expression and adhesive properties and decrease the lung-colonizing potential of HT-29 colon carcinoma cells. Clin Exp Metastasis. 1996;14(2):165–175.
42.
Gilabert-EstellesJRamonLAEspanaF. Expression of angiogenic factors in endometriosis: relationship to fibrinolytic and metalloproteinase systems. Hum Reprod. 2007;22(8):2120–2127.
43.
SotnikovaNYAntsiferovaYSPosiseevaLVShishkovDNPosiseevDVFilippovaES. Mechanisms regulating invasiveness and growth of endometriosis lesions in rat experimental model and in humans. Fertil Steril. 2010;93(8):2701–2705.
44.
ItohHKishoreAHLindqvistARogersDEWordRA. Transforming growth factor beta1 (TGFbeta1) and progesterone regulate matrix metalloproteinases (MMP) in human endometrial stromal cells. J Clin Endocrinol Metab. 2012;97(6):E888–897.
45.
BoettnerBGovekEECrossJVan AelstL. The junctional multidomain protein AF-6 is a binding partner of the Rap1A GTPase and associates with the actin cytoskeletal regulator profilin. Proc Nat Acad Sci U S A. 2000;97(16):9064–9069.
46.
FreyKZivanovicASchwagerKNeriD. Antibody-based targeting of interferon-alpha to the tumor neovasculature: a critical evaluation. Integr Biol. 2011;3(4):468–478.
47.
GhoreschiKThomasPBreitS. Interleukin-4 therapy of psoriasis induces Th2 responses and improves human autoimmune disease. Nat Med. 2003;9(1):40–46.
