Table_1_Proteomic Analysis of Zika Virus Infected Primary Human Fetal Neural Progenitors Suggests a Role for Doublecortin in the Pathological Consequences of Infection in the Cortex.xlsx

Zika virus (ZIKV) infection is associated with severe neurological defects in fetuses and newborns, such as microcephaly. However, the underlying mechanisms remain to be elucidated. In this study, proteomic analysis on ZIKV-infected primary human fetal neural progenitor cells (NPCs) revealed that virus infection altered levels of cellular proteins involved in NPC proliferation, differentiation and migration. The transcriptional levels of some of the altered targets were also confirmed by qRT-PCR. Among the altered proteins, doublecortin (DCX) plays an important role in NPC differentiation and migration. Results showed that ZIKV infection downregulated DCX, at both mRNA and protein levels, as early as 1 day post infection (1 dpi), and lasted throughout the virus replication cycle (4 days). The downregulation of DCX was also observed in a ZIKV-infected fetal mouse brain model, which displayed decreased body weight, brain size and weight, as well as defective cortex structure. By screening the ten viral proteins of ZIKV, we found that both the expression of NS4A and NS5 were correlated with the downregulation of both mRNA and protein levels of DCX in NPCs. These data suggest that DCX is modulated following infection of the brain by ZIKV. How these observed changes of DCX expression translate in the pathological consequences of ZIKV infection and if other cellular proteins are equally involved remains to be investigated.

寨卡病毒（ZIKV）感染与胎儿及新生儿严重的神经系统缺陷相关，如小头畸形。然而，其潜在机制仍有待阐明。在本研究中，对寨卡病毒感染的人原代胎儿神经祖细胞（NPCs）进行的蛋白质组学分析揭示了病毒感染改变了参与NPC增殖、分化和迁移的细胞蛋白水平。通过qRT-PCR也确认了部分改变靶点的转录水平。在改变的蛋白中，双皮质素（DCX）在NPC分化和迁移中起着重要作用。结果显示，ZIKV感染在感染后1天（1 dpi）即下调DCX，并在病毒复制周期（4天）内持续下调。在寨卡病毒感染的胎儿小鼠脑模型中，也观察到DCX的下调，该模型表现出体重减轻、脑体积和重量减少，以及皮质结构缺陷。通过筛选寨卡病毒的十个病毒蛋白，我们发现NS4A和NS5的表达与NPC中DCX的mRNA和蛋白质水平下调相关。这些数据表明，DCX在ZIKV感染后脑部感染的过程中受到调节。DCX表达的这些观察到的变化如何转化为ZIKV感染的病理后果，以及其他细胞蛋白是否同样参与其中，尚需进一步研究。（ZIKV infection is associated with severe neurological defects in fetuses and newborns, such as microcephaly. However, the underlying mechanisms remain to be elucidated. In this study, proteomic analysis on ZIKV-infected primary human fetal neural progenitor cells (NPCs) revealed that virus infection altered levels of cellular proteins involved in NPC proliferation, differentiation and migration. The transcriptional levels of some of the altered targets were also confirmed by qRT-PCR. Among the altered proteins, doublecortin (DCX) plays an important role in NPC differentiation and migration. Results showed that ZIKV infection downregulated DCX, at both mRNA and protein levels, as early as 1 day post infection (1 dpi), and lasted throughout the virus replication cycle (4 days). The downregulation of DCX was also observed in a ZIKV-infected fetal mouse brain model, which displayed decreased body weight, brain size and weight, as well as defective cortex structure. By screening the ten viral proteins of ZIKV, we found that both the expression of NS4A and NS5 were correlated with the downregulation of both mRNA and protein levels of DCX in NPCs. These data suggest that DCX is modulated following infection of the brain by ZIKV. How these observed changes of DCX expression translate in the pathological consequences of ZIKV infection and if other cellular proteins are equally involved remains to be investigated.）