VEGFR-1 blockade disrupts peri-implantation decidual angiogenesis and macrophage recruitment

Vascular Cell

Carrie J Shawber²  Jan K Kitajewski²  Mark V Sauer¹  Chantae S Sullivan-Pyke²  Qian Kun Tan²  Ralf C Zimmermann¹  Nataki C Douglas¹ 

[1] Department of Obstetrics and Gynecology, PH 16–64, Division of Reproductive Endocrinology and Infertility, Columbia University Medical Center, 622 W. 168th Street, New York, NY 10032, USA;Department of Obstetrics and Gynecology, Division of Reproductive Sciences, College of Physicians and Surgeons, Columbia University Medical Center, 630 West 168th St, New York, NY 10032, USA

关键词: Angiogenesis;    Macrophages;    Endothelial cells;    Implantation;    Uterus;    Decidua;    VEGFR-1;   

Others  :  1131027
DOI  :  10.1186/2045-824X-6-16

 received in 2014-03-31, accepted in 2014-07-21,  发布年份 2014

【 摘 要 】

Background

Angiogenesis and macrophage recruitment to the uterus are key features of uterine decidualization; the progesterone-mediated uterine changes that allow for embryo implantation and initiation of pregnancy. In the current study, we characterized the expression of vascular endothelial growth factor receptor-1 (VEGFR-1) in macrophages and endothelial cells of the peri-implantation uterus and determined if VEGFR-1 function is required for decidual angiogenesis, macrophage recruitment, and/or the establishment of pregnancy.

Methods

Expression of VEGFR-1 in uterine endothelial cells and macrophages was determined with immunohistochemistry. To assess the effect of continuous VEGFR-1 blockade, adult female mice were given VEGFR-1 blocking antibody, MF-1, every 3 days for 18 days. After 6 doses, females were mated and a final dose of MF-1 was given on embryonic day 3.5. Endothelial cells and macrophages were quantified on embryonic day 7.5. Pregnancy was analyzed on embryonic days 7.5 and 10.5.

Results

F4/80⁺ macrophages are observed throughout the stroma and are abundant adjacent to the endometrial lumen and glands prior to embryo implantation and scatter throughout the decidua post implantation. VEGFR-1 expression is restricted to the uterine endothelial cells. F4/80⁺ macrophages were often found adjacent to VEGFR-1⁺ endothelial cells in the primary decidual zone. Continuous VEGFR-1 blockade correlates with a significant reduction in decidual vascular and macrophage density, but does not affect embryo implantation or maintenance of pregnancy up to embryonic day 10.5.

Conclusions

We found that VEGFR-1 functions in both decidual angiogenesis and macrophage recruitment to the implantation site during pregnancy. VEGFR-1 is expressed by endothelial cells, however blocking VEGFR-1 function in endothelial cells results in reduced macrophage recruitment to the uterus. VEGFR-1 blockade did not compromise the establishment and/or maintenance of pregnancy.

【 授权许可】

   
2014 Douglas et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Dey SK, Lim H, Das SK, Reese J, Paria BC, Daikoku T, Wang H: Molecular cues to implantation. Endocr Rev 2004, 25:341-373.
• [2]Cha J, Sun X, Dey SK: Mechanisms of implantation: strategies for successful pregnancy. Nat Med 2012, 18:1754-1767.
• [3]Wang H, Dey SK: Roadmap to embryo implantation: clues from mouse models. Nat Rev Genet 2006, 7:185-199.
• [4]Kabawat SE, Mostoufi-Zadeh M, Driscoll SG, Bhan AK: Implantation site in normal pregnancy. A study with monoclonal antibodies. Am J Pathol 1985, 118:76-84.
• [5]Fest S, Aldo PB, Abrahams VM, Visintin I, Alvero A, Chen R, Chavez SL, Romero R, Mor G: Trophoblast-macrophage interactions: a regulatory network for the protection of pregnancy. Am J Reprod Immunol 2007, 57:55-66.
• [6]Rabbani ML, Rogers PA: Role of vascular endothelial growth factor in endometrial vascular events before implantation in rats. Reproduction 2001, 122:85-90.
• [7]Rockwell LC, Pillai S, Olson CE, Koos RD: Inhibition of vascular endothelial growth factor/vascular permeability factor action blocks estrogen-induced uterine edema and implantation in rodents. Biol Reprod 2002, 67:1804-1810.
• [8]Sengupta J, Lalitkumar PG, Najwa AR, Charnock-Jones DS, Evans AL, Sharkey AM, Smith SK, Ghosh D: Immunoneutralization of vascular endothelial growth factor inhibits pregnancy establishment in the rhesus monkey (Macaca mulatta). Reproduction 2007, 133:1199-1211.
• [9]Douglas NC, Tang H, Gomez R, Pytowski B, Hicklin DJ, Sauer CM, Kitajewski J, Sauer MV, Zimmermann RC: Vascular endothelial growth factor receptor 2 (VEGFR-2) functions to promote uterine decidual angiogenesis during early pregnancy in the mouse. Endocrinology 2009, 150:3845-3854.
• [10]Co EC, Gormley M, Kapidzic M, Rosen DB, Scott MA, Stolp HA, McMaster M, Lanier LL, Barcena A, Fisher SJ: Maternal decidual macrophages inhibit NK cell killing of invasive cytotrophoblasts during human pregnancy. Biol Reprod 2013, 88:155.
• [11]Vince GS, Starkey PM, Jackson MC, Sargent IL, Redman CW: Flow cytometric characterisation of cell populations in human pregnancy decidua and isolation of decidual macrophages. J Immunol Methods 1990, 132:181-189.
• [12]Lessin DL, Hunt JS, King CR, Wood GW: Antigen expression by cells near the maternal-fetal interface. Am J Reprod Immunol Microbiol 1988, 16:1-7.
• [13]Wang WJ, Hao CF, Lin QD: Dysregulation of macrophage activation by decidual regulatory T cells in unexplained recurrent miscarriage patients. J Reprod Immunol 2011, 92:97-102.
• [14]Guenther S, Vrekoussis T, Heublein S, Bayer B, Anz D, Knabl J, Navrozoglou I, Dian D, Friese K, Makrigiannakis A, Jeschke U: Decidual macrophages Are significantly increased in spontaneous miscarriages and over-express FasL: a potential role for macrophages in trophoblast apoptosis. Int J Mol Sci 2012, 13:9069-9080.
• [15]Kofler NM, Shawber CJ, Kangsamaksin T, Reed HO, Galatioto J, Kitajewski J: Notch signaling in developmental and tumor angiogenesis. Genes Cancer 2011, 2:1106-1116.
• [16]Turner EC, Hughes J, Wilson H, Clay M, Mylonas KJ, Kipari T, Duncan WC, Fraser HM: Conditional ablation of macrophages disrupts ovarian vasculature. Reproduction 2011, 141:821-831.
• [17]Pipp F, Heil M, Issbrucker K, Ziegelhoeffer T, Martin S, van den Heuvel J, Weich H, Fernandez B, Golomb G, Carmeliet P, Schaper W, Clauss M: VEGFR-1-selective VEGF homologue PlGF is arteriogenic: evidence for a monocyte-mediated mechanism. Circ Res 2003, 92:378-385.
• [18]Murakami M, Zheng Y, Hirashima M, Suda T, Morita Y, Ooehara J, Ema H, Fong GH, Shibuya M: VEGFR1 tyrosine kinase signaling promotes lymphangiogenesis as well as angiogenesis indirectly via macrophage recruitment. Arterioscler Thromb Vasc Biol 2008, 28:658-664.
• [19]Fantin A, Vieira JM, Gestri G, Denti L, Schwarz Q, Prykhozhij S, Peri F, Wilson SW, Ruhrberg C: Tissue macrophages act as cellular chaperones for vascular anastomosis downstream of VEGF-mediated endothelial tip cell induction. Blood 2010, 116:829-840.
• [20]Care AS, Diener KR, Jasper MJ, Brown HM, Ingman WV, Robertson SA: Macrophages regulate corpus luteum development during embryo implantation in mice. J Clin Invest 2013, 123:3472-3487.
• [21]Ferrara N, Gerber HP, LeCouter J: The biology of VEGF and its receptors. Nat Med 2003, 9:669-676.
• [22]Shibuya M: Vascular endothelial growth factor (VEGF) and its receptor (VEGFR) signaling in angiogenesis: a crucial target for anti- and Pro-angiogenic therapies. Genes Cancer 2011, 2:1097-1105.
• [23]Shibuya M: Vascular endothelial growth factor receptor-1 (VEGFR-1/Flt-1): a dual regulator for angiogenesis. Angiogenesis 2006, 9:225-230. discussion 231
• [24]Fong GH, Rossant J, Gertsenstein M, Breitman ML: Role of the Flt-1 receptor tyrosine kinase in regulating the assembly of vascular endothelium. Nature 1995, 376:66-70.
• [25]Hiratsuka S, Minowa O, Kuno J, Noda T, Shibuya M: Flt-1 lacking the tyrosine kinase domain is sufficient for normal development and angiogenesis in mice. Proc Natl Acad Sci 1998, 95:9349-9354.
• [26]Luttun A, Tjwa M, Moons L, Wu Y, Angelillo-Scherrer A, Liao F, Nagy JA, Hooper A, Priller J, De Klerck B, Compernolle V, Daci E, Bohlen P, Dewerchin M, Herbert JM, Fava R, Matthys P, Carmeliet G, Collen D, Dvorak HF, Hicklin DJ, Carmeliet P: Revascularization of ischemic tissues by PlGF treatment, and inhibition of tumor angiogenesis, arthritis and atherosclerosis by anti-Flt1. Nat Med 2002, 8:831-840.
• [27]Wu Y, Zhong Z, Huber J, Bassi R, Finnerty B, Corcoran E, Li H, Navarro E, Balderes P, Jimenez X, Koo H, Mangalampalli VR, Ludwig DL, Tonra JR, Hicklin DJ: Anti-vascular endothelial growth factor receptor-1 antagonist antibody as a therapeutic agent for cancer. Clin Cancer Res 2006, 12:6573-6584.
• [28]Vorontchikhina MA, Zimmermann RC, Shawber CJ, Tang H, Kitajewski J: Unique patterns of Notch1, Notch4 and Jagged1 expression in ovarian vessels during folliculogenesis and corpus luteum formation. Gene Expr Patterns 2005, 5:701-709.
• [29]Kim M, Park HJ, Seol JW, Jang JY, Cho YS, Kim KR, Choi Y, Lydon JP, Demayo FJ, Shibuya M, Ferrara N, Sung HK, Nagy A, Alitalo K, Koh GY: VEGF-A regulated by progesterone governs uterine angiogenesis and vascular remodelling during pregnancy. EMBO Mol Med 2013, 5:1415-1430.
• [30]Brachtendorf G, Kuhn A, Samulowitz U, Knorr R, Gustafsson E, Potocnik AJ, Fassler R, Vestweber D: Early expression of endomucin on endothelium of the mouse embryo and on putative hematopoietic clusters in the dorsal aorta. Dev Dyn 2001, 222:410-419.
• [31]Nakhuda GS, Zimmermann RC, Bohlen P, Liao F, Sauer MV, Kitajewski J: Inhibition of the vascular endothelial cell (VE)-specific adhesion molecule VE-cadherin blocks gonadotropin-dependent folliculogenesis and corpus luteum formation and angiogenesis. Endocrinology 2005, 146:1053-1059.
• [32]Groten T, Fraser HM, Duncan WC, Konrad R, Kreienberg R, Wulff C: Cell junctional proteins in the human corpus luteum: changes during the normal cycle and after HCG treatment. Hum Reprod 2006, 21:3096-3102.
• [33]Rosmarin AG, Weil SC, Rosner GL, Griffin JD, Arnaout MA, Tenen DG: Differential expression of CD11b/CD18 (Mo1) and myeloperoxidase genes during myeloid differentiation. Blood 1989, 73:131-136.
• [34]Keenihan SN, Robertson SA: Diversity in phenotype and steroid hormone dependence in dendritic cells and macrophages in the mouse uterus. Biol Reprod 2004, 70:1562-1572.
• [35]De M, Choudhuri R, Wood GW: Determination of the number and distribution of macrophages, lymphocytes, and granulocytes in the mouse uterus from mating through implantation. J Leukoc Biol 1991, 50:252-262.
• [36]De M, Wood GW: Analysis of the number and distribution of macrophages, lymphocytes, and granulocytes in the mouse uterus from implantation through parturition. J Leukoc Biol 1991, 50:381-392.
• [37]Brandon JM: Macrophage distribution in decidual tissue from early implantation to the periparturient period in mice as defined by the macrophage differentiation antigens F4/80, macrosialin and the type 3 complement receptor. J Reprod Fertil 1995, 103:9-16.
• [38]Paria BC, Zhao X, Das SK, Dey SK, Yoshinaga K: Zonula occludens-1 and E-cadherin are coordinately expressed in the mouse uterus with the initiation of implantation and decidualization. Dev Biol 1999, 208:488-501.
• [39]Chakraborty I, Das SK, Dey SK: Differential expression of vascular endothelial growth factor and its receptor mRNAs in the mouse uterus around the time of implantation. J Endocrinol 1995, 147:339-352.
• [40]Wang ES, Teruya-Feldstein J, Wu Y, Zhu Z, Hicklin DJ, Moore MA: Targeting autocrine and paracrine VEGF receptor pathways inhibits human lymphoma xenografts in vivo. Blood 2004, 104:2893-2902.
• [41]Huang H, Shen J, Vinores SA: Blockade of VEGFR1 and 2 suppresses pathological angiogenesis and vascular leakage in the eye. PLoS One 2011, 6:e21411.
• [42]Carmeliet P, Moons L, Luttun A, Vincenti V, Compernolle V, De Mol M, Wu Y, Bono F, Devy L, Beck H, Scholz D, Acker T, DiPalma T, Dewerchin M, Noel A, Stalmans I, Barra A, Blacher S, VandenDriessche T, Ponten A, Eriksson U, Plate KH, Foidart JM, Schaper W, Charnock-Jones DS, Hicklin DJ, Herbert JM, Collen D, Persico MG: Synergism between vascular endothelial growth factor and placental growth factor contributes to angiogenesis and plasma extravasation in pathological conditions. Nat Med 2001, 7:575-583.
• [43]Fraser HM, Duncan WC: SRB reproduction, fertility and development award lecture 2008. Regulation and manipulation of angiogenesis in the ovary and endometrium. Reprod Fertil Dev 2009, 21:377-392.
• [44]De M, Wood GW: Influence of oestrogen and progesterone on macrophage distribution in the mouse uterus. J Endocrinol 1990, 126:417-424.
• [45]Kirsch TM, Friedman AC, Vogel RL, Flickinger GL: Macrophages in corpora lutea of mice: characterization and effects on steroid secretion. Biol Reprod 1981, 25:629-638.