Unraveling a novel transcription factor code determining the human arterial-specific endothelial cell signature

Blood. 2013 Dec 5;122(24):3982-92. doi: 10.1182/blood-2013-02-483255. Epub 2013 Oct 9.

Abstract

Endothelial cells (ECs) lining arteries and veins have distinct molecular/functional signatures. The underlying regulatory mechanisms are incompletely understood. Here, we established a specific fingerprint of freshly isolated arterial and venous ECs from human umbilical cord comprising 64 arterial and 12 venous genes, representing distinct functions/pathways. Among the arterial genes were 8 transcription factors (TFs), including Notch target HEY2, the current "gold standard" determinant for arterial EC (aEC) specification. Culture abrogated differential gene expression in part due to gradual loss of canonical Notch activity and HEY2 expression. Notably, restoring HEY2 expression or Delta-like4-induced Notch signaling in cultured ECs only partially reinstated the aEC gene signature, whereas combined overexpression of the 8 TFs restored this fingerprint more robustly. Whereas some TFs stimulated few genes, others boosted a large proportion of arterial genes. Although there was some overlap and cross-regulation, the TFs largely complemented each other in regulating the aEC gene profile. Finally, overexpression of the 8 TFs in human umbilical vein ECs conveyed an arterial-like behavior upon their implantation in a Matrigel plug in vivo. Thus, our study shows that Notch signaling determines only part of the aEC signature and identifies additional novel and complementary transcriptional players in the complex regulation of human arteriovenous EC identity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arteries / cytology*
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Blotting, Western
  • Cell Line
  • Cells, Cultured
  • Cluster Analysis
  • Endothelial Cells / metabolism*
  • Gene Regulatory Networks
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Humans
  • Models, Genetic
  • Oligonucleotide Array Sequence Analysis
  • RNA Interference
  • Receptors, Notch / genetics
  • Receptors, Notch / metabolism
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction / genetics
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism
  • Transcriptome*

Substances

  • Basic Helix-Loop-Helix Transcription Factors
  • HEY2 protein, human
  • Receptors, Notch
  • Repressor Proteins
  • Transcription Factors

Associated data

  • GEO/GSE43475