Department of Molecular and Cell Biology, National Center of Biotechnology (CNB-CSIC), Madrid, Spain.
Computational Genomics Service, National Center of Biotechnology (CNB-CSIC), Madrid, Spain.
mBio. 2022 Apr 26;13(2):e0313521. doi: 10.1128/mbio.03135-21. Epub 2022 Mar 1.
Severe acute respiratory syndrome coronavirus (SARS-CoV) and the closely related SARS-CoV-2 are emergent highly pathogenic human respiratory viruses causing acute lethal disease associated with lung damage and dysregulated inflammatory responses. SARS-CoV envelope protein (E) is a virulence factor involved in the activation of various inflammatory pathways. Here, we study the contribution of host miRNAs to the virulence mediated by E protein. Small RNAseq analysis of infected mouse lungs identified miRNA-223 as a potential regulator of pulmonary inflammation, since it was significantly increased in SARS-CoV-WT virulent infection compared to the attenuated SARS-CoV-ΔE infection. inhibition of miRNA-223-3p increased mRNA levels of pro-inflammatory cytokines and inflammasome, suggesting that during lung infection, miRNA-223 might contribute to restrict an excessive inflammatory response. Interestingly, miRNA-223-3p inhibition also increased the levels of the transporter, which is involved in edema resolution and was significantly downregulated in the lungs of mice infected with the virulent SARS-CoV-WT virus. At the histopathological level, a decrease in the pulmonary edema was observed when miR-223-3p was inhibited, suggesting that miRNA-223-3p was involved in the regulation of the SARS-CoV-induced inflammatory pathology. These results indicate that miRNA-223 participates in the regulation of E protein-mediated inflammatory response during SARS-CoV infection by targeting different host mRNAs involved in the pulmonary inflammation, and identify miRNA-223 as a potential therapeutic target in SARS-CoV infection. The SARS-CoV-2 pandemic has emphasized the need to understand the mechanisms of severe lung inflammatory pathology caused by human deadly coronaviruses in order to design new antiviral therapies. Here, we identify miRNA-223-3p as a host miRNA involved in the regulation of lung inflammatory response mediated by envelope (E) protein during SARS-CoV infection. miRNAs downregulate the expression of cellular mRNAs and participate in complex networks of mRNA-miRNA interactions that regulate cellular processes. The inhibition of miRNA-223 in infected mice by intranasal administration of antisense RNAs led to changes in the expression of host factors involved in inflammation (cytokines, chemokines, and inflammasome) and in the resolution of lung edema ion transporter CFTR. These results confirmed the contribution of miRNA-223 to the regulation of SARS-CoV-induced pathogenic processes and support the therapeutic potential of inhibiting miRNAs during coronavirus infection using RNA interference approaches.
严重急性呼吸综合征冠状病毒（SARS-CoV）和密切相关的 SARS-CoV-2 是新兴的高致病性人类呼吸道病毒，可导致与肺部损伤和炎症反应失调相关的急性致死性疾病。SARS-CoV 包膜蛋白（E）是一种与多种炎症途径激活有关的毒力因子。在这里，我们研究了宿主 microRNA 对 E 蛋白介导的毒力的贡献。感染小鼠肺部的小 RNAseq 分析鉴定 microRNA-223 是一种潜在的肺部炎症调节剂，因为与减毒 SARS-CoV-ΔE 感染相比，它在 SARS-CoV-WT 毒力感染中显著增加。miRNA-223-3p 的抑制增加了促炎细胞因子和炎性小体的 mRNA 水平，这表明在肺部感染期间，miRNA-223 可能有助于限制过度的炎症反应。有趣的是，miRNA-223-3p 的抑制也增加了转运蛋白的水平，该转运蛋白参与水肿的消退，并且在感染 SARS-CoV-WT 病毒的小鼠肺部显著下调。在组织病理学水平上，当抑制 miR-223-3p 时，观察到肺水肿减少，表明 miR-223-3p 参与调节 SARS-CoV 诱导的炎症病理学。这些结果表明，miRNA-223 通过靶向参与肺部炎症的不同宿主 mRNAs，参与 SARS-CoV 感染过程中介导 E 蛋白的炎症反应调节，并将 miRNA-223 鉴定为 SARS-CoV 感染的潜在治疗靶点。SARS-CoV-2 大流行强调了需要了解人类致命冠状病毒引起的严重肺部炎症病理的机制，以便设计新的抗病毒疗法。在这里，我们确定了 microRNA-223-3p 是一种宿主 microRNA，它参与调节 SARS-CoV 感染过程中介导包膜（E）蛋白的肺部炎症反应。miRNA 下调细胞 mRNA 的表达，并参与调节细胞过程的复杂 mRNA-miRNA 相互作用网络。通过鼻内给予反义 RNA 抑制感染小鼠中的 miRNA-223 导致参与炎症的宿主因子（细胞因子、趋化因子和炎性小体）和肺水肿离子转运蛋白 CFTR 的表达发生变化。这些结果证实了 miRNA-223 对调节 SARS-CoV 诱导的致病过程的贡献，并支持使用 RNA 干扰方法抑制冠状病毒感染期间的 microRNAs 的治疗潜力。
