Wingless (Wnt)-3A induces trophoblast migration and matrix metalloproteinase-2 secretion through canonical Wnt signaling and protein kinase B/AKT activation.

Invasion of human trophoblasts is promoted through activation of wingless (Wnt) signaling, suggesting a role of the pathway in placental development and morphogenesis. However, details on the process such as involvement of canonical and/or noncanonical Wnt signaling cascades as well as their target genes are largely unknown. Hence, signal transduction via canonical Wnt signaling or phosphatidylinositide 3-kinase (PI3K)/AKT and their cross talk as well as trophoblast-specific protease expression were investigated in trophoblastic SGHPL-5 cells and primary extravillous trophoblasts purified from first-trimester placentas. Western blot analyses revealed that the recombinant Wnt ligand Wnt-3A increased phosphorylation of AKT and the downstream kinase glycogen synthase kinase (GSK)-3beta as well as accumulation of activated, nuclear beta-catenin. In accordance, luciferase expression of a canonical Wnt/TCF reporter and cell migration in first-trimester villous explant cultures and of SGHPL-5 cells were stimulated. Chemical inhibition of PI3K abolished Wnt-dependent phosphorylation of AKT and GSK-3beta and trophoblast motility but did not affect appearance of activated beta-catenin or Wnt/TCF reporter activity. In contrast, inhibition of the canonical pathway through soluble Dickkopf-1 did not influence AKT and GSK-3beta phosphorylation but reduced Wnt reporter activity, accumulation of active beta-catenin, and cell migration. Both inhibitors decreased Wnt-3A-induced secretion of pro- and active matrix metalloproteinase-2 from SGHPL-5 cells and pure EVT. The data suggest that Wnt-3A may activate canonical Wnt signaling and PI3K/AKT through distinct receptors. The two signaling cascades act independently in trophoblasts; however, both pathways promote Wnt-dependent migration and the release of matrix metalloproteinase-2, which has been identified as novel Wnt target in invasive trophoblasts.

[1]  D. Pant,et al.  Lymphoid Enhancer Factor 1‐Mediated Wnt Signaling Promotes the Initiation of Trophoblast Lineage Differentiation in Mouse Embryonic Stem Cells , 2008, Stem cells.

[2]  L. Leyns,et al.  Non‐conventional Frizzled ligands and Wnt receptors , 2008, Development, growth & differentiation.

[3]  G. Burton,et al.  Gestational diseases -- a workshop report. , 2008, Placenta.

[4]  H. Husslein,et al.  Human chorionic gonadotropin stimulates trophoblast invasion through extracellularly regulated kinase and AKT signaling. , 2008, Endocrinology.

[5]  E. Duan,et al.  Dickkopf-1 secreted by decidual cells promotes trophoblast cell invasion during murine placentation. , 2008, Reproduction.

[6]  Adam J Pawson,et al.  Nuclear stabilization of beta-catenin and inactivation of glycogen synthase kinase-3beta by gonadotropin-releasing hormone: targeting Wnt signaling in the pituitary gonadotrope. , 2007, Molecular endocrinology.

[7]  M. Knöfler,et al.  Human chorionic gonadotrophin attenuates NF-kappaB activation and cytokine expression of endometriotic stromal cells. , 2007, Molecular human reproduction.

[8]  C. Leisser,et al.  Complex expression pattern of Wnt ligands and frizzled receptors in human placenta and its trophoblast subtypes. , 2007, Placenta.

[9]  Sung-Eun Kim,et al.  The PI3 kinase-Akt pathway mediates Wnt3a-induced proliferation. , 2007, Cellular signalling.

[10]  Beibei Wu,et al.  Wnt signaling induces matrix metalloproteinase expression and regulates T cell transmigration. , 2007, Immunity.

[11]  R. Nusse,et al.  Wnt Signaling: Multiple Pathways, Multiple Receptors, and Multiple Transcription Factors* , 2006, Journal of Biological Chemistry.

[12]  J. Tseng,et al.  Differential expression of growth-, angiogenesis- and invasion-related factors in the development of placenta accreta. , 2006, Taiwanese journal of obstetrics & gynecology.

[13]  M. Bilban,et al.  Activation of the canonical wingless/T-cell factor signaling pathway promotes invasive differentiation of human trophoblast. , 2006, The American journal of pathology.

[14]  L. Giudice,et al.  Dickkopf-1, an inhibitor of Wnt signaling, is regulated by progesterone in human endometrial stromal cells. , 2006, The Journal of clinical endocrinology and metabolism.

[15]  A. Basak,et al.  Role of pro-IGF-II processing by proprotein convertase 4 in human placental development. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[16]  Hiroyuki Aburatani,et al.  Phosphatidylinositol 3-Kinase–Akt Pathway Plays a Critical Role in Early Cardiomyogenesis by Regulating Canonical Wnt Signaling , 2005, Circulation research.

[17]  D. Dufort,et al.  Uterine Wnt/β-catenin signaling is required for implantation , 2005 .

[18]  J. Pollheimer,et al.  Signalling pathways regulating the invasive differentiation of human trophoblasts: a review. , 2005, Placenta.

[19]  S. Jeon,et al.  Both ERK and Wnt/β-catenin pathways are involved in Wnt3a-induced proliferation , 2005, Journal of Cell Science.

[20]  B. Tsang,et al.  EGF-induced trophoblast secretion of MMP-9 and TIMP-1 involves activation of both PI3K and MAPK signalling pathways. , 2004, Reproduction.

[21]  B. Tsang,et al.  Both mitogen-activated protein kinase and phosphatidylinositol 3-kinase signalling are required in epidermal growth factor-induced human trophoblast migration. , 2004, Molecular human reproduction.

[22]  P. Husslein,et al.  Transcriptional regulation of the human chorionic gonadotropin beta gene during villous trophoblast differentiation. , 2004, Endocrinology.

[23]  L. Giudice,et al.  Identification, characterization, and regulation of the canonical Wnt signaling pathway in human endometrium. , 2003, The Journal of clinical endocrinology and metabolism.

[24]  Judith E. Cartwright,et al.  Hepatocyte growth factor induced human trophoblast motility involves phosphatidylinositol-3-kinase, mitogen-activated protein kinase, and inducible nitric oxide synthase. , 2002, Experimental cell research.

[25]  C. Jomary,et al.  Secreted Frizzled-related proteins: searching for relationships and patterns. , 2002, BioEssays : news and reviews in molecular, cellular and developmental biology.

[26]  R. Glazer,et al.  Akt1 induces extracellular matrix invasion and matrix metalloproteinase-2 activity in mouse mammary epithelial cells. , 2001, Cancer research.

[27]  M. Cybulsky,et al.  Wnt7b regulates placental development in mice. , 2001, Developmental biology.

[28]  P. Balaram,et al.  Demonstration of TGF-alpha-EGFR and EGF-EGFR autocrine loops and their relation to proliferation in complete hydatidiform moles (CHM). , 2001, International journal of gynecological cancer : official journal of the International Gynecological Cancer Society.

[29]  M. Vidaud,et al.  Characterization of Human Villous and Extravillous Trophoblasts Isolated from First Trimester Placenta , 2001, Laboratory Investigation.

[30]  S. Fukumoto,et al.  Akt Participation in the Wnt Signaling Pathway through Dishevelled* , 2001, The Journal of Biological Chemistry.

[31]  B. Hogan,et al.  Cellular and molecular responses of the uterus to embryo implantation can be elicited by locally applied growth factors. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[32]  F. McCormick,et al.  Differential Regulation of Glycogen Synthase Kinase 3β by Insulin and Wnt Signaling* , 2000, The Journal of Biological Chemistry.

[33]  Choy My,et al.  Efficient, rapid and reliable establishment of human trophoblast cell lines using poly-L-ornithine. , 2000 .

[34]  A. Campana,et al.  Paracrine and autocrine regulators of trophoblast invasion--a review. , 2000, Placenta.

[35]  Dianqing Wu,et al.  Suppression of Glycogen Synthase Kinase Activity Is Not Sufficient for Leukemia Enhancer Factor-1 Activation* , 1999, The Journal of Biological Chemistry.

[36]  J. Christiansen,et al.  Targeted disruption of the Wnt2 gene results in placentation defects. , 1996, Development.

[37]  R. Pijnenborg,et al.  Placental bed spiral arteries in the hypertensive disorders of pregnancy , 1991, British journal of obstetrics and gynaecology.

[38]  J. Bland,et al.  Uteroplacental arterial changes related to interstitial trophoblast migration in early human pregnancy. , 1983, Placenta.

[39]  A. Weeraratna,et al.  Assaying Wnt5A-mediated invasion in melanoma cells. , 2008, Methods in molecular biology.

[40]  E. Norwitz.  Defective implantation and placentation: laying the blueprint for pregnancy complications. , 2007, Reproductive biomedicine online.

[41]  E. Staun-ram,et al.  Reproductive Biology and Endocrinology Open Access Expression and Importance of Matrix Metalloproteinase 2 and 9 (mmp-2 and -9) in Human Trophoblast Invasion , 2004 .

[42]  M. Kühl.  The WNT/calcium pathway: biochemical mediators, tools and future requirements. , 2004, Frontiers in bioscience : a journal and virtual library.

[43]  I. Manyonda,et al.  Efficient, rapid and reliable establishment of human trophoblast cell lines using poly-L-ornithine. , 2000, Early pregnancy.

[44]  M. Depew,et al.  Wnt3a−/−-like phenotype and limb deficiency in Lef1−/−Tcf1−/− mice , 1999 .

[45]  G. Hamilton,et al.  Growth factors, proteases and protease inhibitors in the maternal-fetal dialogue. , 1996, Placenta.