Distinct responses to in vitro ZIKV infection of iPSC-derived trophoblasts from dizygotic twin babies discordant for congenital Zika syndrome
Murilo Sena Amaral1, Alessandra Soares Shanoski1, Luiz Carlos de Caires Junior2, Ernesto Goulart2, David Abraham Morales-Vicente1, Renato Astray1, Jorge Elias Kalil Filho3, Mayana Zatz2, Sergio Verjovski-Almeida1,4*
1Butantan Institute, São Paulo, Brazil; 2Human Genome and Stem Cell Research Center, Biosciences Institute, University of São Paulo (USP), São Paulo, Brazil; 3Medical School and Heart Institute, University of São Paulo (USP), São Paulo, Brazil; 4Institute of Chemistry, University of São Paulo (USP), São Paulo, Brazil
Zika virus (ZIKV) is a flavivirus that has been associated with severe abnormalities in newborns, leading to congenital Zika syndrome (CZS). We have recently found that neural progenitor cells from discordant CZS twins show differential in vitro susceptibility to ZIKV, but it remains to be determined if the trophoblasts from these twins and, ultimately, the placenta, also play a role in the different ZIKV infection outcomes. Here, we obtained blood from three pairs of dizygotic twins discordant for CZS (one affected, one non-affected), reprogrammed their erythroblasts towards human induced pluripotent stem cells (hiPSCs), induced them to differentiate into trophoblasts and compared the in vitro trophoblast's gene expression profile and the outcome of ZIKV infection. We observed that ZIKV replicates significantly more in hiPSC-derived trophoblasts from affected (CZS) twins than in the non-affected counterparts. In addition, upon ZIKV in vitro infection, the trophoblasts from the non-affected twins are able to secrete higher amounts of a set of pro-inflammatory cytokines, as compared to the affected ones. Transcriptome profiling revealed a significant (q-value < 0.05) enrichment of Gene Ontology (GO) terms among the differentially expressed genes between the trophoblasts from CZS-affected and non-affected individuals after ZIKV infection, including “integrin binding”, “fibronectin binding” as well as “blood circulation” among the GOs of downregulated genes in trophoblasts from CZS-affected twins. These results indicate that trophoblasts from the non-affected twins are able to respond more efficiently to ZIKV infection than trophoblasts from affected (CZS) twins.
Work funded by grants from FAPESP # 2014/03620-2 to SVA, and 2013/08028-1 and 14/50931-3 and from INCT/CNPq # 465355/2014-5 to MZ.