Evaluation of protein expression and phosphorylation of SARS-CoV-2-infected placental tissue in the second trimester by tandem mass tag system
Evaluation of protein expression and phosphorylation of SARS-CoV-2-infected placental tissue in the second trimester by tandem mass tag system
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DOI：
10.1016/j.jri.2023.103889
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发表时间：
2023-03-21
影响因子：
3.4
通讯作者：
Hayakawa, Satoshi
中科院分区：
文献类型：
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作者：
Takada, Kazuhide;Trinh, Quang Duy;Komine-Aizawa, Shihoko;Tominaga, Daisuke;Tsuji, Noriko M.;Yamada, Hideto;Hayakawa, Satoshi
目的：尽管偶尔在人胎盘组织中检测到 SARS-CoV-2 的病毒基因组或蛋白质，但子宫内垂直 SARS-CoV-2 感染相当罕见。为了揭示胎盘屏障抑制病毒传播给胎儿的机制，我们检测了从SARS-CoV-2感染母亲收集的胎盘样本中的蛋白质表达和磷酸化。方法：在伦理委员会的批准下，我们从SARS-CoV-2阳性母亲那里获取胎盘样本。通过实时 PCR 证实胎盘中存在 SARS-CoV-2 感染。我们比较了患有轻度 COVID-19 的母亲产下的两个感染 SARS-CoV-2 的胎盘（双胞胎）和患有严重 COVID-19 的母亲产下的 1 个未感染的胎盘。这三名妇女均在妊娠中期通过剖腹产分娩。我们还分析了从宫内胎儿死亡 (IUFD) 妇女身上采集的三个胎盘样本。所有患者均在妊娠中期感染 SARS-CoV-2。通过串联质量标签 (TMT) 系统检查冷冻样品，该系统能够区分样品之间的蛋白质并进行相对定量，包括其磷酸化水平。结果：TMT 系统检测到 2675 个蛋白质和 778 个磷酸化肽。与未感染的胎盘相比，在感染的胎盘中，包括先天性和获得性免疫系统在内的几个重要途径均上调。这些途径的磷酸化水平也被激活。补体级联、VEGF 信号传导和 RHO GTPase 信号传导也发生了变化。在IUFD患者中，胎盘中仍富集SARS-CoV-2感染相关通路，但不能排除死后组织降解的影响。结论：妊娠期SARS-CoV-2感染影响胎盘蛋白表达和磷酸化。除了直接的细胞毒性外，SARS-CoV-2 感染还必须引起胎盘的组织病理学和功能变化。对胎盘屏障的局部免疫反应和机制的分析能够挽救受感染母亲的胎盘功能障碍以及随后的胎儿健康。
Purpose: Vertical SARS-CoV-2 infection in utero is quite rare, although the viral genome or proteins of SARS-CoV-2 are occasionally detected in human placental tissue. To reveal the mechanism of the placental barrier that inhibits viral transmission to the fetus, we examined protein expression and phosphorylation in placental samples collected from SARS-CoV-2-infected mothers.Methods: Under the approval of the ethical committees, we obtained placental samples from SARS-CoV-2-positive mothers. SARS-CoV-2 infection in the placentae was confirmed by real-time PCR. We compared two SARS-CoV-2-infected placentae (twins) delivered from a mother with mild COVID-19 and 1 uninfected placenta from a mother with severe COVID-19. All three of these women delivered by cesarean section during the second trimester. We also analyzed three placental samples collected from women with intrauterine fetal death (IUFD). All patients were infected with SARS-CoV-2 during the second trimester. Frozen samples were examined by a tandem mass tag (TMT) system, which enabled the differentiation and relative quantification of proteins between samples, including their phosphorylation levels.Results: The TMT system detected 2675 proteins and 778 phosphorylated peptides. Compared with the uninfected placenta, in the infected placentae, several essential pathways, including the innate and acquired immune systems, were upregulated. The phosphorylation levels of these pathways were also activated. The complement cascade, VEGF signaling, and RHO GTPase signaling were also changed. In patients with IUFD, SARS-CoV-2 infection-related pathways were still enriched in the placentae, while the effects of postmortem tissue degradation could not be ruled out.Conclusion: SARS-CoV-2 infection during pregnancy affects placental protein expression and phosphorylation. SARS-CoV-2 infection must cause both histopathological and functional changes in the placenta in addition to direct cytotoxicity. Analysis of the local immune responses and mechanisms of the placental barrier enables the rescue of placental dysfunction in infected mothers and subsequent fetal well-being.