A Grhl2-dependent gene network controls trophoblast branching morphogenesis

Development. 2015 Mar 15;142(6):1125-36. doi: 10.1242/dev.113829.

Abstract

Healthy placental development is essential for reproductive success; failure of the feto-maternal interface results in pre-eclampsia and intrauterine growth retardation. We found that grainyhead-like 2 (GRHL2), a CP2-type transcription factor, is highly expressed in chorionic trophoblast cells, including basal chorionic trophoblast (BCT) cells located at the chorioallantoic interface in murine placentas. Placentas from Grhl2-deficient mouse embryos displayed defects in BCT cell polarity and basement membrane integrity at the chorioallantoic interface, as well as a severe disruption of labyrinth branching morphogenesis. Selective Grhl2 inactivation only in epiblast-derived cells rescued all placental defects but phenocopied intraembryonic defects observed in global Grhl2 deficiency, implying the importance of Grhl2 activity in trophectoderm-derived cells. ChIP-seq identified 5282 GRHL2 binding sites in placental tissue. By integrating these data with placental gene expression profiles, we identified direct and indirect Grhl2 targets and found a marked enrichment of GRHL2 binding adjacent to genes downregulated in Grhl2(-/-) placentas, which encoded known regulators of placental development and epithelial morphogenesis. These genes included that encoding the serine protease inhibitor Kunitz type 1 (Spint1), which regulates BCT cell integrity and labyrinth formation. In human placenta, we found that human orthologs of murine GRHL2 and its targets displayed co-regulation and were expressed in trophoblast cells in a similar domain as in mouse placenta. Our data indicate that a conserved Grhl2-coordinated gene network controls trophoblast branching morphogenesis, thereby facilitating development of the site of feto-maternal exchange. This might have implications for syndromes related to placental dysfunction.

Keywords: Basement membrane defects; Epithelial differentiation; Epithelial morphogenesis; Placenta defects; Spint1.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites / genetics
  • Chromatin Immunoprecipitation
  • DNA-Binding Proteins / metabolism*
  • Female
  • Fluorescent Antibody Technique
  • Gene Regulatory Networks / genetics
  • Gene Regulatory Networks / physiology*
  • Humans
  • Immunohistochemistry
  • Microarray Analysis
  • Microscopy, Electron
  • Morphogenesis / physiology*
  • Placentation*
  • Pregnancy
  • Proteinase Inhibitory Proteins, Secretory / genetics
  • Real-Time Polymerase Chain Reaction
  • Transcription Factors / metabolism*
  • Trophoblasts / physiology*

Substances

  • DNA-Binding Proteins
  • GRHL2 protein, human
  • Proteinase Inhibitory Proteins, Secretory
  • SPINT1 protein, human
  • Transcription Factors

Associated data

  • GEO/GSE65960
  • GEO/GSE65962
  • GEO/GSE65963