Betacoronaviruses: activation and antagonism of host innate immune responses

β冠状病毒：宿主先天免疫反应的激活和拮抗

• 批准号: 10735058
• 负责人: Susan R Weiss
• 金额: $ 54.66万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-24 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10735058
• 关键词: 2019-nCoV; Biology; COVID-19 pneumonia; Cell Death; Cells; Coronavirus; Cyclic AMP-Dependent Protein Kinases; Data; Disease; Disease Outbreaks; Double-Stranded RNA; Endoribonucleases; Feedback; Future; Genes; Genome; Goals; Human; Immune; Immune response; In Vitro; Individual; Infection; Inflammation; Inflammatory; Innate Immune Response; Interferons; Ligase; Lung; Messenger RNA; Middle East Respiratory Syndrome; Middle East Respiratory Syndrome Coronavirus; Mus; Outcome; Pathogenicity; Pathway interactions; Pattern recognition receptor; Peptide Initiation Factors; Pharmaceutical Preparations; Phosphorylation; Phosphotransferases; Pneumonia; Production; Protein Biosynthesis; Protein Dephosphorylation; Proteins; RNA; Respiratory Disease; Ribonucleases; Role; SARS coronavirus; SARS-CoV-2 infection; Sarbecovirus; Scheme; Signal Transduction; Structure; System; Therapeutic; Upper respiratory tract; Viral; Viral Genome; Viral Proteins; Virus; Virus Replication; Work; antagonist; attenuation; betacoronavirus; biological adaptation to stress; cell injury; combat; coronavirus treatment; endoplasmic reticulum stress; experience; human coronavirus; in vivo; interest; lung injury; mouse model; mutant; novel therapeutic intervention; oligoadenylate; phosphoric diester hydrolase; prevent; protein activation; protein kinase R; replicase; respiratory; response; reverse genetics; severe COVID-19; virus host interaction

Human betacoronaviruses induce a wide spectrum of respiratory disease. MERS-CoV (MERS) and SARS-CoV- 2 (SARS-2), cause severe lethal pneumonia, while SARS-2 can also cause mild to asymptomatic disease in some individuals. In contrast, infections with the “common” respiratory human beta-CoV OC43 are largely limited to the upper respiratory tract. With only a few drugs that show promise against human CoVs, it remains imperative to develop therapeutics for current and future emergent human CoVs. For this, we need to identify and understand the host-virus interactions common to all or different among these viruses. A major focus of our lab has been on the double-stranded (ds)RNA induced host responses to beta-CoVs: interferon production and signaling; oligoadenylate synthetase-ribonuclease L and protein kinase R (PKR), that are both antiviral and pro- inflammatory. We have shown that the conserved CoV replicase encoded nsp15 endoribonuclease (Endo)U reduces dsRNA accumulation and activation of all three pathways while subgenera specific accessory proteins add to the antagonism. Merbeco (MERS-CoV) and embeco (OC43) viruses shut down these pathways effectively; in contrast, the sarbecovirus, SARS-2, replicates in respiratory derived cells despite activation of all three. Preliminary data suggest that downstream effects of RNase L on inflammation and cell death may be more impactful during SARS-2 infection than the modest antiviral effects. PKR and PKR-like ER kinase (PERK), both kinases of the integrated stress response, when activated, phosphorylate protein synthesis initiation factor eIF2a, leading to protein synthesis attenuation, providing an important control point in infection. Activation of PERK also initiates a pathways of the unfolded protein response (UPR) to ER stress, which produces antiviral effects and inflammation. We found that MERS and OC43 infection suppress phosphorylation of eIF2a by preventing PKR activation and through a PERK pathway feedback loop, dephosphorylating eIF2a. In contrast SARS-2 promotes p-eIF2a through both PKR and PERK, but maintains viral protein synthesis. This may occur in part via the CoV replicase nsp1 protein, which promotes selective attenuation of host, but not viral, protein synthesis. Our overall hypothesis is that betacoronaviruses of each subgenus have evolved unique ways to interact with host responses to optimize replication and spread and that these differences influence pathogenic outcomes. We propose to determine: the role of the conserved CoV EndoU and subgenera specific accessory proteins in antagonizing host innate response pathways; the impact of OAS-RNase L activation on viral replication, inflammation and cell death; and the effects of host PKR/PERK and viral nsp1 on control of protein synthesis and virus replication. Our overall goal is to understand the similarities and differences in host interactions among beta-CoVs of three subgenera. Our findings may identify novel therapeutic interventions targeting host or viral proteins and indicate whether they may work as pan-CoV therapies or be unique to each beta-CoV subgenus, and thus help prepare for future CoV outbreaks.

人贝曲纳病毒会诱导广泛的呼吸道疾病。 MERS-COV（MERS）和SARS-COV- 2（SARS-2），引起严重的致命性肺炎，而SARS-2也会引起轻度至无症状的疾病 一些人。相比之下，“常见”呼吸道人β-COV OC43的感染在很大程度上受到限制 到上呼吸道。只有几种药物表现出对人类cov的承诺，它仍然是 必须为当前和未来的新兴人类COV开发理论。为此，我们需要确定 并了解这些病毒中所有或不同的宿主病毒相互作用。我们的主要重点 实验室一直处于双链（DS）RNA诱导的宿主对β-COV的反应：干扰素产生和 信号传导；寡核苷酸合成酶 - 核酸酶L和蛋白激酶R（PKR），既是抗病毒又是抗病毒 炎症。我们已经证明了配置的COV复制酶编码的NSP15内核核酸酶（Endo）U 减少DSRNA的积累和所有三个途径的激活，而亚属特异性附件蛋白 加入对抗。 Merbeco（MERS-COV）和Embeco（OC43）病毒关闭了这些途径 有效地;相反，SARBECOVIRUS，SARS-2在呼吸衍生细胞的目的地激活中复制 三。初步数据表明，RNase L对注射和细胞死亡的下游影响可能更多 SARS-2感染期间的影响比适度的抗病毒作用。 PKR和PKR样ER激酶（PERK），都 综合应力反应的激酶，当激活时，磷酸化蛋白质合成启动因子EIF2A， 导致蛋白质合成衰减，从而在感染中提供了重要的控制点。也激活特权 启动展开的蛋白质反应（UPR）对ER应激的途径，这会产生抗病毒作用和 炎。我们发现MER和OC43感染通过预防PKR抑制EIF2A的磷酸化 激活和通过PERK途径反馈回路，脱磷酸化EIF2A。相反，SARS-2促进 P-EIF2A通过PKR和PERK，但保持病毒蛋白的合成。这可能部分通过COV发生 复制酶NSP1蛋白，可促进宿主的选择性衰减，但不是病毒蛋白合成。我们的整体 假设是，每个子属的贝塔曲霉病毒已经发展出与宿主互动的独特方法 对优化复制和扩散的反应，这些差异会影响致病结果。 我们建议确定：在 对抗主机先天反应途径； OAS-RNase L激活对病毒复制的影响， 炎症和细胞死亡；以及宿主PKR/PERK和病毒NSP1对蛋白质合成的控制的影响 和病毒复制。我们的总体目标是了解主机互动之间的相似性和差异 三个亚属的β蛋白盒。我们的发现可能确定针对宿主或病毒的新型热干预措施 蛋白质并指示它们是否可以作为泛蛋白疗法工作或对每个β-COV子属独有， 因此，有助于为将来的COV爆发做准备。

项目成果

SARS-CoV-2 Delta Variant (AY.3) in the Feces of a Domestic Cat.

• DOI: 10.3390/v14020421
• 发表时间: 2022-02-17
• 期刊: Viruses
• 作者: Lenz OC;Marques AD;Kelly BJ;Rodino KG;Cole SD;Perera RAPM;Weiss SR;Bushman FD;Lennon EM
• 通讯作者: Lennon EM

SARS-CoV-2 infection, neuropathogenesis and transmission among deer mice: Implications for reverse zoonosis to New World rodents.

• DOI: 10.1101/2020.08.07.241810
• 发表时间: 2020-08-07
• 期刊: bioRxiv : the preprint server for biology
• 作者: Fagre, Anna;Lewis, Juliette;Schountz, Tony
• 通讯作者: Schountz, Tony

SARS-CoV-2 infection, neuropathogenesis and transmission among deer mice: Implications for spillback to New World rodents.

• DOI: 10.1371/journal.ppat.1009585
• 发表时间: 2021-05
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Fagre A;Lewis J;Eckley M;Zhan S;Rocha SM;Sexton NR;Burke B;Geiss B;Peersen O;Bass T;Kading R;Rovnak J;Ebel GD;Tjalkens RB;Aboellail T;Schountz T
• 通讯作者: Schountz T

A Novel Glucocorticoid and Androgen Receptor Modulator Reduces Viral Entry and Innate Immune Inflammatory Responses in the Syrian Hamster Model of SARS-CoV-2 Infection.

• DOI: 10.3389/fimmu.2022.811430
• 发表时间: 2022
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Rocha SM;Fagre AC;Latham AS;Cummings JE;Aboellail TA;Reigan P;Aldaz DA;McDermott CP;Popichak KA;Kading RC;Schountz T;Theise ND;Slayden RA;Tjalkens RB
• 通讯作者: Tjalkens RB

Partial ORF1ab Gene Target Failure with Omicron BA.2.12.1.

• DOI: 10.1128/jcm.00600-22
• 发表时间: 2022-06-15
• 期刊: Journal of clinical microbiology
• 影响因子: 9.4