PDF(Pubmed)
Abstract：
Human cytomegalovirus (CMV) infection is the leading non-genetic cause of congenital malformation in developed countries, causing significant fetal injury, and in some cases fetal death. The pathogenetic mechanisms through which this host-specific virus infects then damages both the placenta and the fetal brain are currently ill-defined. We investigated the CMV modulation of key signaling pathway proteins for these organs including dual-specificity tyrosine phosphorylation-regulated kinases (DYRK) and Sonic Hedgehog (SHH) pathway proteins using human first trimester placental trophoblast (TEV-1) cells, primary human astrocyte (NHA) brain cells, and CMV-infected human placental tissue. Immunofluorescence demonstrated the accumulation and re-localization of SHH proteins in CMV-infected TEV-1 cells with Gli2, Ulk3, and Shh re-localizing to the CMV cytoplasmic virion assembly complex (VAC). In CMV-infected NHA cells, DYRK1A re-localized to the VAC and DYRK1B re-localized to the CMV nuclear replication compartments, and the SHH proteins re-localized with a similar pattern as was observed in TEV-1 cells. Western blot analysis in CMV-infected TEV-1 cells showed the upregulated expression of Rb, Ulk3, and Shh, but not Gli2. In CMV-infected NHA cells, there was an upregulation of DYRK1A, DYRK1B, Gli2, Rb, Ulk3, and Shh. These in vitro monoculture findings are consistent with patterns of protein upregulation and re-localization observed in naturally infected placental tissue and CMV-infected ex vivo placental explant histocultures. This study reveals CMV-induced changes in proteins critical for fetal development, and identifies new potential targets for CMV therapeutic development.
摘要：
人类巨细胞病毒（CMV）感染是发达国家先天性畸形的主要非遗传原因，造成严重的胎儿伤害，在某些情况下还有胎儿死亡.这种宿主特异性病毒感染然后损害胎盘和胎儿大脑的致病机制目前尚不明确。我们使用人类早孕胎盘滋养层（TEV-1）细胞研究了这些器官的关键信号通路蛋白的CMV调节，包括双特异性酪氨酸磷酸化调节激酶（DYRK）和SonicHedgehog（SHH）通路蛋白。原代人星形胶质细胞（NHA）脑细胞，和CMV感染的人胎盘组织。免疫荧光显示SHH蛋白在CMV感染的TEV-1细胞中的积累和重新定位，Gli2，Ulk3和Shh重新定位到CMV细胞质病毒体组装复合物（VAC）。在CMV感染的NHA细胞中，DYRK1A重新定位到VAC，DYRK1B重新定位到CMV核复制区室，并且SHH蛋白以与TEV-1细胞中观察到的相似的模式重新定位。在CMV感染的TEV-1细胞中的蛋白质印迹分析显示Rb的表达上调,Ulk3和嘘，但不是Gli2。在CMV感染的NHA细胞中，有DYRK1A的上调，DYRK1B,Gli2,Rb,Ulk3和嘘。这些体外单培养发现与在自然感染的胎盘组织和CMV感染的离体胎盘外植体组织培养中观察到的蛋白质上调和重新定位的模式一致。这项研究揭示了CMV诱导的对胎儿发育至关重要的蛋白质变化，并确定CMV治疗开发的新潜在靶标。
