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

Murilo Sena Amaral,Mayana Zatz,Jorge E Kalil,David Abraham Morales-Vicente,Alessandra Soares-Schanoski,Luiz Carlos Caires-Júnior,Roselane Paiva Gomes,Ernesto Goulart,Sergio Verjovski-Almeida,Giovanna Gonçalves De Oliveira Olberg,Renato Mancini Astray,Kristy O Murray

doi:10.1371/journal.pntd.0008424

Murilo Sena Amaral, Mayana Zatz + Show 10 more

Open Access

https://doi.org/10.1371/journal.pntd.0008424

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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.

Highlights

Zika virus (ZIKV) is a flavivirus with sporadic outbreaks reported in several countries, causing an infection usually characterized by mild symptoms, where up to 80% of cases remain asymptomatic [1,2,3]
It is estimated that congenital Zika syndrome (CZS) occurs in ~1–40% of cases of pregnant women infected by ZIKV, which suggests that different susceptibility factors might be involved, including the host genetic background
By analyzing trophoblast cells that recapitulate the placenta from three pairs of dizygotic twins discordant for CZS, we were able to show that trophoblasts from CZS-affected twins were significantly more susceptible to ZIKV infection when compared with trophoblasts from the non-affected twins

Summary

Introduction

Zika virus (ZIKV) is a flavivirus with sporadic outbreaks reported in several countries, causing an infection usually characterized by mild symptoms, where up to 80% of cases remain asymptomatic [1,2,3]. ZIKV in the maternal circulation needs to cross the placental barrier that includes fetal endothelial cells and trophoblasts in order to reach the fetus [23,24]. Several placenta related cells have been shown to be infected by ZIKV, including placental macrophages and trophoblasts [25,26,27,28,29]

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