title: Endothelin-1 down-regulates matrix metalloproteinase 14 and 15 expression in human first trimester trophoblasts via endothelin receptor type B.
abstract: Does endothelin-1 (ET-1) regulate matrix metalloproteinase (MMP) 14 and 15 production and invasion of human first trimester trophoblasts?
  
  
  
  ET-1 in pathophysiological concentrations down-regulates MMP14 and MMP15 expression via endothelin receptor (ETR) type B and decreases trophoblast migration and invasion.
  
  
  
  MMP14 and MMP15 are involved in trophoblast invasion. Impairment of invasion has been linked to pregnancy complications such as pre-eclampsia (PE). ET-1 is up-regulated in PE.
  
  
  
  In vitro study using primary human trophoblasts from 50 first trimester placentas (gestational week 7-12).
  
  
  
  Trophoblasts were cultured in the absence or presence of 10-100 nM ET-1. MMP14 and MMP15 mRNA and protein were quantified by RT-qPCR and Western blotting, respectively. Selective antagonists for ETRA (BQ-123) or ETRB (BQ-788) were used to identify ETR subtypes involved. Functional ET-1 effects were tested in first trimester chorionic villous explants and transwell invasion assays. The roles of tumor necrosis factor (TNF)-α (25 ng/ml) and oxygen (1%) in ET-1 regulation of MMP14 and 15 expression were assessed by Western blotting.
  
  
  
  ET-1 down-regulated MMP14 and MMP15 mRNA (-21% and -26%, respectively, P < 0.05) and protein levels (-18% and -22%, respectively, P < 0.05). This effect was mediated via ETRB. ET-1 decreased trophoblast outgrowth in placental explants (-24%, P < 0.05) and trophoblast invasion (-26%, P ≤ 0.01). TNF-α enhanced ET-1 mediated MMP15 down-regulation (by 10%, P < 0.05), whereas hypoxia abolished the effect of ET-1 on both MMPs.
  
  
  
  N/A.
  
  
  
  Only primary trophoblasts were used in this study. Since trophoblast yield from first trimester placental material is limited, further aspects of MMP14 and 15 regulation could not be characterized. Other anti-invasive factors may be altered by ET-1 in trophoblasts and, thus, contribute to the reduced invasion, but have not been investigated. Oxygen levels similar to those found in the decidua (5-8% O2) were not analyzed in this study.
  
  
  
  ET-1 modifies placental function already during the first trimester of pregnancy, the time-window when the placental changes implicated in PE occur. Thus, our results improve the understanding of the placental mechanisms underlying trophoblast invasion and PE.
  
  
  
  The study was funded by the Oesterreichische Nationalbank (Anniversary Fund, project number: 14796) and the Herzfelder'sche Familienstiftung (to J.P.; number: 00685). AMM received funding from the Austrian Science Fund FWF (W1241) and the Medical University Graz through the PhD Program Molecular Fundamentals of Inflammation (DK-MOLIN). The authors have no conflict of interest.
authors.affiliation: Department of Obstetrics and Gynecology, Medical University of Graz, Auenbruggerplatz 14, Graz 8036, Austria.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Majali-Martinez
authors.affiliation: Department of Obstetrics and Fetal-Maternal Medicine, Medical University of Vienna, Währinger Gürtel 18-20, Vienna 1090, Austria.
authors.forename: Philipp
authors.initials: P
authors.lastname: Velicky
authors.affiliation: Department of Obstetrics and Fetal-Maternal Medicine, Medical University of Vienna, Währinger Gürtel 18-20, Vienna 1090, Austria.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Pollheimer
authors.affiliation: Department of Obstetrics and Fetal-Maternal Medicine, Medical University of Vienna, Währinger Gürtel 18-20, Vienna 1090, Austria.
authors.forename: Martin
authors.initials: M
authors.lastname: Knöfler
authors.affiliation: Department of Physiology, Development and Neuroscience, Centre for Trophoblast Research, University of Cambridge, Downing Street, Cambridge CB2 3 EG, UK.
authors.forename: Hong Wa
authors.initials: HW
authors.lastname: Yung
authors.affiliation: Department of Physiology, Development and Neuroscience, Centre for Trophoblast Research, University of Cambridge, Downing Street, Cambridge CB2 3 EG, UK.
authors.forename: Graham J
authors.initials: GJ
authors.lastname: Burton
authors.affiliation: Institute of Pathophysiology and Immunology, SFL Chicken CAM Lab, Medical University of Graz, Heinrichstrasse 31a, Graz 8010, Austria.
authors.forename: Nassim Ghaffari
authors.initials: NG
authors.lastname: Tabrizi-Wizsy
authors.affiliation: Department of Obstetrics and Gynecology, Medical University of Graz, Auenbruggerplatz 14, Graz 8036, Austria.
authors.forename: Uwe
authors.initials: U
authors.lastname: Lang
authors.affiliation: Department of Obstetrics and Gynecology, Medical University of Graz, Auenbruggerplatz 14, Graz 8036, Austria.
authors.forename: Ursula
authors.initials: U
authors.lastname: Hiden
authors.affiliation: Department of Obstetrics and Gynecology, Medical University of Graz, Auenbruggerplatz 14, Graz 8036, Austria.
authors.forename: Gernot
authors.initials: G
authors.lastname: Desoye
authors.affiliation: Department of Obstetrics and Gynecology, Medical University of Graz, Auenbruggerplatz 14, Graz 8036, Austria.
authors.forename: Martina
authors.initials: M
authors.lastname: Dieber-Rotheneder
mesh_terms: D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects*; D019332:Endothelin-1 / Q000494:pharmacology*; D005260:Female; D006801:Humans; D053511:Matrix Metalloproteinase 14 / Q000235:genetics / Q000378:metabolism*; D053517:Matrix Metalloproteinase 15 / Q000235:genetics / Q000378:metabolism*; D010920:Placenta / Q000187:drug effects / Q000378:metabolism; D011247:Pregnancy; D011261:Pregnancy Trimester, First / Q000378:metabolism; D044023:Receptor, Endothelin B / Q000235:genetics / Q000378:metabolism*; D014327:Trophoblasts / Q000187:drug effects* / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: C000588596:EDNRB protein, human; D019332:Endothelin-1; C507098:MMP15 protein, human; D044023:Receptor, Endothelin B; D014409:Tumor Necrosis Factor-alpha; D053517:Matrix Metalloproteinase 15; C507086:MMP14 protein, human; D053511:Matrix Metalloproteinase 14
keywords: MMPs; endothelin-1; hypoxia; inflammation; invasion; pre-eclampsia; trophoblast