title: p45 NF-E2 regulates syncytiotrophoblast differentiation by post-translational GCM1 modifications in human intrauterine growth restriction.
abstract: Placental insufficiency jeopardizes prenatal development, potentially leading to intrauterine growth restriction (IUGR) and stillbirth. Surviving fetuses are at an increased risk for chronic diseases later in life. IUGR is closely linked with altered trophoblast and placental differentiation. However, due to a paucity of mechanistic insights, suitable biomarkers and specific therapies for IUGR are lacking. The transcription factor p45 NF-E2 (nuclear factor erythroid derived 2) has been recently found to regulate trophoblast differentiation in mice. The absence of p45 NF-E2 in trophoblast cells causes IUGR and placental insufficiency in mice, but mechanistic insights are incomplete and the relevance of p45 NF-E2 for human syncytiotrophoblast differentiation remains unknown. Here we show that p45 NF-E2 negatively regulates human syncytiotrophoblast differentiation and is associated with IUGR in humans. Expression of p45 NF-E2 is reduced in human placentae complicated with IUGR compared with healthy controls. Reduced p45 NF-E2 expression is associated with increased syncytiotrophoblast differentiation, enhanced glial cells missing-1 (GCM1) acetylation and GCM1 desumoylation in IUGR placentae. Induction of syncytiotrophoblast differentiation in BeWo and primary villous trophoblast cells with 8-bromo-adenosine 3',5'-cyclic monophosphate (8-Br-cAMP) reduces p45 NF-E2 expression. Of note, p45 NF-E2 knockdown is sufficient to increase syncytiotrophoblast differentiation and GCM1 expression. Loss of p45 NF-E2 using either approach resulted in CBP-mediated GCM1 acetylation and SENP-mediated GCM1 desumoylation, demonstrating that p45 NF-E2 regulates post-translational modifications of GCM1. Functionally, reduced p45 NF-E2 expression is associated with increased cell death and caspase-3 activation in vitro and in placental tissues samples. Overexpression of p45 NF-E2 is sufficient to repress GCM1 expression, acetylation and desumoylation, even in 8-Br-cAMP exposed BeWo cells. These results suggest that p45 NF-E2 negatively regulates differentiation and apoptosis activation of human syncytiotrophoblast by modulating GCM1 acetylation and sumoylation. These studies identify a new pathomechanism related to IUGR in humans and thus provide new impetus for future studies aiming to identify new biomarkers and/or therapies of IUGR.
authors.affiliation: Institute of Clinical Chemistry and Pathobiochemistry, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany.|Health Campus Immunology, Infectiology and Inflammation, Otto-von-Guericke-University, Magdeburg, Germany.
authors.forename: Shrey
authors.initials: S
authors.lastname: Kohli
authors.affiliation: Institute of Clinical Chemistry and Pathobiochemistry, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany.|Health Campus Immunology, Infectiology and Inflammation, Otto-von-Guericke-University, Magdeburg, Germany.
authors.forename: Juliane
authors.initials: J
authors.lastname: Hoffmann
authors.affiliation: Institute of Clinical Chemistry and Pathobiochemistry, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany.|Health Campus Immunology, Infectiology and Inflammation, Otto-von-Guericke-University, Magdeburg, Germany.
authors.forename: Franziska
authors.initials: F
authors.lastname: Lochmann
authors.affiliation: Institute of Clinical Chemistry and Pathobiochemistry, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany.|Health Campus Immunology, Infectiology and Inflammation, Otto-von-Guericke-University, Magdeburg, Germany.
authors.forename: Paulina
authors.initials: P
authors.lastname: Markmeyer
authors.affiliation: Department of Gynaecology and Obstetrics, Comprehensive Cancer Center Erlangen-FMN, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany.
authors.forename: Hanna
authors.initials: H
authors.lastname: Huebner
authors.affiliation: Department of Pediatrics and Adolescent Medicine, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany.
authors.forename: Fabian B
authors.initials: FB
authors.lastname: Fahlbusch
authors.affiliation: Institute of Clinical Chemistry and Pathobiochemistry, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany.|Health Campus Immunology, Infectiology and Inflammation, Otto-von-Guericke-University, Magdeburg, Germany.
authors.forename: Moh'd Mohanad
authors.initials: MM
authors.lastname: Al-Dabet
authors.affiliation: Institute of Clinical Chemistry and Pathobiochemistry, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany.|Health Campus Immunology, Infectiology and Inflammation, Otto-von-Guericke-University, Magdeburg, Germany.
authors.forename: Ihsan
authors.initials: I
authors.lastname: Gadi
authors.affiliation: Institute of Clinical Chemistry and Pathobiochemistry, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany.|Health Campus Immunology, Infectiology and Inflammation, Otto-von-Guericke-University, Magdeburg, Germany.
authors.forename: Jayakumar
authors.initials: J
authors.lastname: Manoharan
authors.affiliation: Gynecology and Obstetrics, Klinikum Magdeburg, Magdeburg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Löttge
authors.affiliation: Experimental Obstetrics and Gynecology, Medical Faculty, Otto-von-Guericke University, Magdeburg, Germany.
authors.forename: Ana C
authors.initials: AC
authors.lastname: Zenclussen
authors.affiliation: Department of Hematology, Rambam Health Care Campus, Haifa, Israel.
authors.forename: Anat
authors.initials: A
authors.lastname: Aharon
authors.affiliation: Department of Hematology, Rambam Health Care Campus, Haifa, Israel.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Brenner
authors.affiliation: Institute of Clinical Chemistry and Pathobiochemistry, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany.|Health Campus Immunology, Infectiology and Inflammation, Otto-von-Guericke-University, Magdeburg, Germany.|Department of Biotechnology, University of Sargodha, Pakistan.
authors.forename: Khurrum
authors.initials: K
authors.lastname: Shahzad
authors.affiliation: Department of Gynaecology and Obstetrics, Comprehensive Cancer Center Erlangen-FMN, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Ruebner
authors.affiliation: Institute of Clinical Chemistry and Pathobiochemistry, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany.|Health Campus Immunology, Infectiology and Inflammation, Otto-von-Guericke-University, Magdeburg, Germany.
authors.forename: Berend
authors.initials: B
authors.lastname: Isermann
mesh_terms: D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000107:Acetylation / Q000187:drug effects; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D004268:DNA-Binding Proteins; D005260:Female; D005317:Fetal Growth Retardation / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D051379:Mice; D050984:NF-E2 Transcription Factor, p45 Subunit / Q000235:genetics / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D011247:Pregnancy; D011499:Protein Processing, Post-Translational; D058207:Sumoylation / Q000187:drug effects / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014327:Trophoblasts / Q000378:metabolism* / Q000473:pathology
chemical_list: D004268:DNA-Binding Proteins; C491089:GCM1 protein, human; D050984:NF-E2 Transcription Factor, p45 Subunit; C494692:NFE2 protein, human; D009687:Nuclear Proteins; D014157:Transcription Factors; D015124:8-Bromo Cyclic Adenosine Monophosphate; C505407:CASP3 protein, human; D053148:Caspase 3