Human Cytomegalovirus Dysregulates Cellular Dual-Specificity Tyrosine Phosphorylation-Regulated Kinases and Sonic Hedgehog Pathway Proteins in Neural Astrocyte and Placental Models

Viruses. 2024 Jun 5;16(6):918. doi: 10.3390/v16060918.


Human cytomegalovirus (CMV) infection is the leading non-genetic cause of congenital malformation in developed countries, causing significant fetal injury, and in some cases fetal death. The pathogenetic mechanisms through which this host-specific virus infects then damages both the placenta and the fetal brain are currently ill-defined. We investigated the CMV modulation of key signaling pathway proteins for these organs including dual-specificity tyrosine phosphorylation-regulated kinases (DYRK) and Sonic Hedgehog (SHH) pathway proteins using human first trimester placental trophoblast (TEV-1) cells, primary human astrocyte (NHA) brain cells, and CMV-infected human placental tissue. Immunofluorescence demonstrated the accumulation and re-localization of SHH proteins in CMV-infected TEV-1 cells with Gli2, Ulk3, and Shh re-localizing to the CMV cytoplasmic virion assembly complex (VAC). In CMV-infected NHA cells, DYRK1A re-localized to the VAC and DYRK1B re-localized to the CMV nuclear replication compartments, and the SHH proteins re-localized with a similar pattern as was observed in TEV-1 cells. Western blot analysis in CMV-infected TEV-1 cells showed the upregulated expression of Rb, Ulk3, and Shh, but not Gli2. In CMV-infected NHA cells, there was an upregulation of DYRK1A, DYRK1B, Gli2, Rb, Ulk3, and Shh. These in vitro monoculture findings are consistent with patterns of protein upregulation and re-localization observed in naturally infected placental tissue and CMV-infected ex vivo placental explant histocultures. This study reveals CMV-induced changes in proteins critical for fetal development, and identifies new potential targets for CMV therapeutic development.

Keywords: astrocytes; cytomegalovirus; developmental pathways; pathogenesis; placenta.

MeSH terms

  • Astrocytes* / metabolism
  • Astrocytes* / virology
  • Cell Line
  • Cells, Cultured
  • Cytomegalovirus Infections* / metabolism
  • Cytomegalovirus Infections* / virology
  • Cytomegalovirus* / physiology
  • Dyrk Kinases
  • Female
  • Hedgehog Proteins* / genetics
  • Hedgehog Proteins* / metabolism
  • Humans
  • Phosphorylation
  • Placenta* / metabolism
  • Placenta* / virology
  • Pregnancy
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism
  • Protein-Tyrosine Kinases* / genetics
  • Protein-Tyrosine Kinases* / metabolism
  • Signal Transduction*
  • Trophoblasts / metabolism
  • Trophoblasts / virology


  • Hedgehog Proteins
  • Protein-Tyrosine Kinases
  • Protein Serine-Threonine Kinases
  • SHH protein, human
  • Dyrk Kinases

Grants and funding

This study was supported by grants from the Thelma Pyne Postdoctoral Award (Hamilton—PS64847) and the Australia–Germany Joint Research Cooperation Scheme (RG192195).