Differential gene expression elicited by ZIKV infection in trophoblasts from congenital Zika syndrome discordant twins

PLoS Negl Trop Dis. 2020 Aug 3;14(8):e0008424. doi: 10.1371/journal.pntd.0008424. eCollection 2020 Aug.

Abstract

Zika virus (ZIKV) causes congenital Zika syndrome (CZS), which is characterized by fetal demise, microcephaly and other abnormalities. ZIKV in the pregnant woman circulation must cross the placental barrier that includes fetal endothelial cells and trophoblasts, in order to reach the fetus. CZS occurs in ~1-40% of cases of pregnant women infected by ZIKV, suggesting that mothers' infection by ZIKV during pregnancy is not deterministic for CZS phenotype in the fetus. Therefore, other susceptibility factors might be involved, including the host genetic background. We have previously shown that in three pairs of dizygotic twins discordant for CZS, neural progenitor cells (NPCs) from the CZS-affected twins presented differential in vitro ZIKV susceptibility compared with NPCs from the non-affected. Here, we analyzed human-induced-pluripotent-stem-cell-derived (hiPSC-derived) trophoblasts from these twins and compared by RNA-Seq the trophoblasts from CZS-affected and non-affected twins. Following in vitro exposure to a Brazilian ZIKV strain (ZIKVBR), trophoblasts from CZS-affected twins were significantly more susceptible to ZIKVBR infection when compared with trophoblasts from the non-affected. Transcriptome profiling revealed no differences in gene expression levels of ZIKV candidate attachment factors, IFN receptors and IFN in the trophoblasts, either before or after ZIKVBR infection. Most importantly, ZIKVBR infection caused, only in the trophoblasts from CZS-affected twins, the downregulation of genes related to extracellular matrix organization and to leukocyte activation, which are important for trophoblast adhesion and immune response activation. In addition, only trophoblasts from non-affected twins secreted significantly increased amounts of chemokines RANTES/CCL5 and IP10 after infection with ZIKVBR. Overall, our results showed that trophoblasts from non-affected twins have the ability to more efficiently activate genes that are known to play important roles in cell adhesion and in triggering the immune response to ZIKV infection in the placenta, and this may contribute to predict protection from ZIKV dissemination into fetuses' tissues.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chemokines / metabolism
  • Extracellular Matrix
  • Female
  • Gene Expression*
  • Genetic Predisposition to Disease
  • Humans
  • Induced Pluripotent Stem Cells
  • Infant
  • Pregnancy
  • Pregnancy Complications, Infectious / genetics
  • Pregnancy Complications, Infectious / virology
  • Trophoblasts / metabolism*
  • Trophoblasts / virology
  • Twins, Dizygotic*
  • Zika Virus
  • Zika Virus Infection / congenital*
  • Zika Virus Infection / genetics

Substances

  • Chemokines

Grants and funding

This work was supported by grants from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) (Thematic grant number 2014/03620-2 and 2018/23693-5 to SVA and CEPID number 2013/08028-1 and INCT number 14/50931-3 to MZ) and from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) INCT number 465355/2014-5 to MZ. DAMV received a fellowship from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Brazil – Finance Code 001; LCCJ is a fellow of FAPESP (2017/16283-2); EG is a fellow of FAPESP (2019/18469-1); SVA laboratory was also supported by institutional funds from Fundação Butantan; SVA received an established investigator fellowship award from CNPq, Brazil. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.