Teratogenic Rubella Virus Alters the Endodermal Differentiation Capacity of Human Induced Pluripotent Stem Cells
Teratogenic Rubella Virus Alters the Endodermal Differentiation Capacity of Human Induced Pluripotent Stem Cells
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DOI：
10.3390/cells8080870
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发表时间：
2019-08-01
期刊：
影响因子：
6
通讯作者：
Claus, Claudia
中科院分区：
文献类型：
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作者：
Bilz, Nicole C.;Willscher, Edith;Claus, Claudia
人类先天性病毒感染的研究需要合适的离体平台来研究胚胎发育过程中的物种特异性事件。这些感染的一个突出例子是风疹病毒（RV），它最常导致耳朵、心脏和眼睛发育缺陷。我们将致畸性RV应用于人诱导多能干细胞（iPSCs），然后将其分化为三种胚胎谱系（外胚层、中胚层和内胚层），作为囊胚和原肠胚样阶段的细胞培养模型。在RV存在的情况下，谱系特异性分化标记得以表达，表明谱系身份得以维持。然而，对所有样本的转录组表达特征的肖像分析显示，模拟病毒和rv感染的内胚层细胞之间的相关性低于它们的外胚层和中胚层细胞。RV感染期间，确定内胚层标记物增加。表观遗传格局发生了深刻的变化，包括染色质重塑复合物组分的表达水平和III型干扰素的诱导，特别是在rv感染的ipsc内胚层分化后。此外，视野转录因子RAX和SIX3以及血管发生基因组的组成部分被鉴定为转录异常。尽管iPSC形态得以维持，但RV感染过程中，胚胎样体作为三维细胞聚集体的形成和细胞粘附能力受到损害。在iPSCs分化过程中，RV诱导的分子改变与先天性风疹综合征临床体征的相关性提示了病毒损害人类发育的机制。
The study of congenital virus infections in humans requires suitable ex vivo platforms for the species-specific events during embryonal development. A prominent example for these infections is rubella virus (RV) which most commonly leads to defects in ear, heart, and eye development. We applied teratogenic RV to human induced pluripotent stem cells (iPSCs) followed by differentiation into cells of the three embryonic lineages (ecto-, meso-, and endoderm) as a cell culture model for blastocyst- and gastrulation-like stages. In the presence of RV, lineage-specific differentiation markers were expressed, indicating that lineage identity was maintained. However, portrait analysis of the transcriptomic expression signatures of all samples revealed that mock- and RV-infected endodermal cells were less related to each other than their ecto- and mesodermal counterparts. Markers for definitive endoderm were increased during RV infection. Profound alterations of the epigenetic landscape including the expression level of components of the chromatin remodeling complexes and an induction of type III interferons were found, especially after endodermal differentiation of RV-infected iPSCs. Moreover, the eye field transcription factors RAX and SIX3 and components of the gene set vasculogenesis were identified as dysregulated transcripts. Although iPSC morphology was maintained, the formation of embryoid bodies as three-dimensional cell aggregates and as such cellular adhesion capacity was impaired during RV infection. The correlation of the molecular alterations induced by RV during differentiation of iPSCs with the clinical signs of congenital rubella syndrome suggests mechanisms of viral impairment of human development.