Zika virus (ZIKV) is largely known for causing brain abnormalities due to its ability to infect neural progenitor stem cells during early development. Here, we show that ZIKV is also capable of infecting and destroying stem-like cancer cells from aggressive human embryonal tumors of the central nervous system (CNS). When evaluating the oncolytic properties of Brazilian Zika virus strain (ZIKV) against human breast, prostate, colorectal, and embryonal CNS tumor cell lines, we verified a selective infection of CNS tumor cells followed by massive tumor cell death. ZIKV was more efficient in destroying embryonal CNS tumorspheres than normal stem cell neurospheres. A single intracerebroventricular injection of ZIKV in BALB/c nude mice bearing orthotopic human embryonal CNS tumor xenografts resulted in a significantly longer survival, decreased tumor burden, fewer metastasis, and complete remission in some animals. Tumor cells closely resembling neural stem cells at the molecular level with activated Wnt signaling were more susceptible to the oncolytic effects of ZIKV Furthermore, modulation of Wnt signaling pathway significantly affected ZIKV-induced tumor cell death and viral shedding. Altogether, these preclinical findings indicate that ZIKV could be an efficient agent to treat aggressive forms of embryonal CNS tumors and could provide mechanistic insights regarding its oncolytic effects. Brazilian Zika virus strain kills aggressive metastatic forms of human CNS tumors and could be a potential oncolytic agent for cancer therapy. .

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 (ZIKV BR), trophoblasts from CZS-affected twins were significantly more susceptible to ZIKV BR 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 ZIKV BR infection. Most importantly, ZIKV BR 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 ZIKV BR. 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 PLOS NEGLECTED TROPICAL DISEASES

Please find enclosed the manuscript entitled "Differential gene expression elicited by ZIKV infection in trophoblasts from congenital Zika syndrome discordant twins", by Amaral et al., which we are submitting to PLOS Pathogens as a research article. In this manuscript, we report for the first time the different susceptibilities to ZIKV in vitro infection of human-inducedpluripotent-stem-cell-derived (hiPSC-derived) trophoblasts from three pairs of twins discordant for congenital Zika syndrome (CZS).Discordant twins represent a good case-control group to test for the genetic contribution determining the fetuses' outcome of gestational infection. Each of the twins' expectant mother has been infected during pregnancy, the two fetuses have been exposed to ZIKV at the same time during their embryonic development, and yet only one of them presented a CZS phenotype. To the best of our knowledge, no other study has compared the in vitro trophoblast gene expression profile or outcome of ZIKV infection in hiPSC-derived trophoblasts from discordant twins for CZS under the same experimental conditions.We have previously shown that, in these three pairs of dizygotic twins discordant for CZS, the neural progenitor cells (NPCs) from the CZS-affected twins presented higher in vitro ZIKV susceptibility compared with NPCs from the non-affected (Caires-Junior et al., Nat Commun 9: 475, 2018). Here, we show that trophoblasts from CZS-affected twins were also significantly more susceptible to ZIKV infection when compared with trophoblasts from the non-affected. In addition, ZIKV 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. Notably, only trophoblasts from non-affected twins secreted significantly increased amounts of chemokines RANTES/CCL5 and IP10 after infection with ZIKV.Taken together, these results show that development of congenital Zika syndrome might result among other factors from a concomitant decreased ability of the placenta to respond to ZIKV infection in the CZS-affected neonates, along with a deregulation of neural development genes in ZIKV-infected NPCs of these CZS-affected neonates. We are positive that this report will be of great interest to the readers of PLOS Pathogens and to those working in the field.We would like to suggest one of the following editors to handle our submission: Michael S. Diamond or Ana Fernandez-Sesma.We are looking forward to hearing from you. Yours sincerely, Abstract 29Zika virus (ZIKV) causes congenital Zika syndrome (CZS), which is characterized by fetal 30 demise, microcephaly and other abnormalities. ZIKV in the pregnant woman circulation must cross 31 the placental barrier that includes fetal endothelial cells and trophoblasts, in order to reach the fetus. 32 CZS occurs in ~1-40% of cases of pregnant women infected by ZIKV, suggesting that mothers' 33 infection by ZIKV during pregnancy...

<p>Molecular characterization and stem cell properties of USP7-ATRT cell line.</p>

<p>Description of supplementary materials & methods and supplementary figure legends.</p>