Department of Microbiology and Medical Zoology, School of Medicine, University of Puerto Rico Medical Sciences Campus, San Juan, PR, USA.
Department of Biology, University of Puerto Rico Rio Piedras Campus, San Juan, PR, USA.
J Mol Histol. 2022 Apr;53(2):199-214. doi: 10.1007/s10735-021-09994-w. Epub 2021 Jul 15.
Zika virus (ZIKV) infection has been associated with fetal abnormalities by compromising placental integrity, but the mechanisms by which this occurs are unknown. Flavivirus can deregulate the host proteome, especially extracellular matrix (ECM) proteins. We hypothesize that a deregulation of specific ECM proteins by ZIKV, affects placental integrity. Using twelve different placental samples collected during the 2016 ZIKV Puerto Rico epidemic, we compared the proteome of five ZIKV infected samples with four uninfected controls followed by validation of most significant proteins by immunohistochemistry. Quantitative proteomics was performed using tandem mass tag TMT10plex™ Isobaric Label Reagent Set followed by Q Exactive™ Hybrid Quadrupole Orbitrap Mass Spectrometry. Identification of proteins was performed using Proteome Discoverer 2.1. Proteins were compared based on the fold change and p value using Limma software. Significant proteins pathways were analyzed using Ingenuity Pathway (IPA). TMT analysis showed that ZIKV infected placentas had 94 reviewed differentially abundant proteins, 32 more abundant, and 62 less abundant. IPA analysis results indicate that 45 of the deregulated proteins are cellular components of the ECM and 16 play a role in its structure and organization. Among the most significant proteins in ZIKV positive placenta were fibronectin, bone marrow proteoglycan, and fibrinogen. Of these, fibrinogen was further validated by immunohistochemistry in 12 additional placenta samples and found significantly increased in ZIKV infected placentas. The upregulation of this protein in the placental tissue suggests that ZIKV infection is promoting the coagulation of placental tissue and restructuration of ECM potentially affecting the integrity of the tissue and facilitating dissemination of the virus from mother to the fetus.
寨卡病毒(ZIKV)感染通过损害胎盘完整性与胎儿畸形有关,但具体机制尚不清楚。黄病毒可使宿主蛋白质组失调,特别是细胞外基质(ECM)蛋白。我们假设 ZIKV 对特定 ECM 蛋白的失调会影响胎盘的完整性。我们使用在 2016 年波多黎各寨卡病毒流行期间收集的 12 个不同的胎盘样本,比较了 5 个 ZIKV 感染样本和 4 个未感染对照样本的蛋白质组,随后通过免疫组织化学验证了大多数显著蛋白。使用串联质量标签 TMT10plex™同重同位素标记试剂试剂盒进行定量蛋白质组学分析,然后进行 Q Exactive™杂交四极杆轨道阱质谱分析。使用 Proteome Discoverer 2.1 进行蛋白质鉴定。使用 Limma 软件基于倍数变化和 p 值比较蛋白质。使用 IPA(Ingenuity Pathway)分析显著蛋白质途径。TMT 分析显示,ZIKV 感染的胎盘有 94 个经审查的差异丰度蛋白,其中 32 个蛋白丰度增加,62 个蛋白丰度减少。IPA 分析结果表明,失调蛋白中有 45 个是细胞外基质的细胞成分,16 个蛋白在其结构和组织中发挥作用。在 ZIKV 阳性胎盘的最显著蛋白中,有纤连蛋白、骨髓蛋白聚糖和纤维蛋白原。其中,纤维蛋白原在另外 12 个胎盘样本中通过免疫组织化学进一步验证,在 ZIKV 感染的胎盘组织中发现其显著增加。该蛋白在胎盘组织中的上调表明,ZIKV 感染正在促进胎盘组织的凝血和 ECM 的重构,这可能影响组织的完整性,并促进病毒从母亲传播到胎儿。