Gata4 regulates hedgehog signaling and Gata6 expression for outflow tract development

PLoS Genet. 2019 May 23;15(5):e1007711. doi: 10.1371/journal.pgen.1007711. eCollection 2019 May.

Abstract

Dominant mutations of Gata4, an essential cardiogenic transcription factor (TF), were known to cause outflow tract (OFT) defects in both human and mouse, but the underlying molecular mechanism was not clear. In this study, Gata4 haploinsufficiency in mice was found to result in OFT defects including double outlet right ventricle (DORV) and ventricular septum defects (VSDs). Gata4 was shown to be required for Hedgehog (Hh)-receiving progenitors within the second heart field (SHF) for normal OFT alignment. Restored cell proliferation in the SHF by knocking-down Pten failed to rescue OFT defects, suggesting that additional cell events under Gata4 regulation is important. SHF Hh-receiving cells failed to migrate properly into the proximal OFT cushion, which is associated with abnormal EMT and cell proliferation in Gata4 haploinsufficiency. The genetic interaction of Hh signaling and Gata4 is further demonstrated to be important for OFT development. Gata4 and Smo double heterozygotes displayed more severe OFT abnormalities including persistent truncus arteriosus (PTA). Restoration of Hedgehog signaling renormalized SHF cell proliferation and migration, and rescued OFT defects in Gata4 haploinsufficiency. In addition, there was enhanced Gata6 expression in the SHF of the Gata4 heterozygotes. The Gata4-responsive repressive sites were identified within 1kbp upstream of the transcription start site of Gata6 by both ChIP-qPCR and luciferase reporter assay. These results suggested a SHF regulatory network comprising of Gata4, Gata6 and Hh-signaling for OFT development.

Publication types

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

MeSH terms

  • Animals
  • Cell Movement
  • Cell Proliferation
  • Embryo, Mammalian
  • GATA4 Transcription Factor / genetics*
  • GATA4 Transcription Factor / metabolism
  • GATA6 Transcription Factor / genetics*
  • GATA6 Transcription Factor / metabolism
  • Gene Expression Regulation
  • Haploinsufficiency
  • Hedgehog Proteins / genetics*
  • Hedgehog Proteins / metabolism
  • Heterozygote
  • Humans
  • Mice
  • Mice, Transgenic
  • Myocytes, Cardiac / metabolism
  • Myocytes, Cardiac / pathology
  • Signal Transduction
  • Smoothened Receptor / genetics*
  • Smoothened Receptor / metabolism
  • Truncus Arteriosus / abnormalities
  • Truncus Arteriosus / metabolism
  • Ventricular Outflow Obstruction / genetics*
  • Ventricular Outflow Obstruction / metabolism
  • Ventricular Outflow Obstruction / pathology
  • Ventricular Septum / metabolism*
  • Ventricular Septum / pathology

Substances

  • GATA4 Transcription Factor
  • GATA6 Transcription Factor
  • Gata4 protein, mouse
  • Gata6 protein, mouse
  • Hedgehog Proteins
  • Shh protein, mouse
  • Smo protein, mouse
  • Smoothened Receptor