Endothelial cell tropism in the pathogenesis and host response against influenza viruses - Resubmission 01

流感病毒发病机制和宿主反应中的内皮细胞向性 - 重新提交 01

• 批准号: 10303023
• 负责人: Balaji Manicassamy
• 金额: $ 54.03万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-08 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10303023
• 关键词: Acute Respiratory Distress Syndrome; Adult; Antiviral Response; Attenuated Vaccines; Avian Influenza A Virus; Biological Models; Blood Vessels; Cell Adhesion Molecules; Cells; Characteristics; Coculture Techniques; Development; Disease; Disease Progression; Down-Regulation; Elderly; Endothelial Cells; Endothelium; Engineering; Epidemic; Epithelial Cells; Extravasation; Ferrets; Generations; Goals; Health; Hematopoietic; Human; Immune; Immune response; Immunocompromised Host; Individual; Infection; Inflammatory; Inflammatory Response; Influenza A Virus, H1N1 Subtype; Influenza A Virus, H5N1 Subtype; Influenza A Virus, H7N7 Subtype; Influenza A Virus, H7N9 Subtype; Influenza A virus; Injury; Innate Immune Response; Integration Host Factors; Knowledge; Link; Lower respiratory tract structure; Lung; Mediating; MicroRNAs; Modeling; Molecular; Morbidity - disease rate; Mus; Nature; Neutrophil Infiltration; Nose; Pathogenesis; Pathogenicity; Positioning Attribute; Predisposition; Production; Pulmonary Edema; Reporter; Reporting; Research Personnel; Role; Severity of illness; Site; Survival Rate; Symptoms; Testing; Therapeutic; Transgenic Organisms; Tropism; Upper Respiratory Infections; Vascular Permeabilities; Viral; Viral Genes; Viral Genome; Viral Load result; Viral Pathogenesis; Viral Pneumonia; Virulence; Virulent; Virus; Virus Diseases; Virus Replication; Vulnerable Populations; age group; burden of illness; cell type; cytokine; experimental study; immunopathology; improved; in vivo; influenza infection; influenza virus strain; influenzavirus; lung injury; member; monocyte; mortality; mouse model; neutrophil; novel; novel therapeutic intervention; pandemic disease; pathogen; prevent; rational design; recombinant virus; recruit; seasonal influenza; tissue tropism; tool

ABSTRACT Influenza A viruses pose a serious threat to human health, causing seasonal epidemics and occasional pandemics that result in significant morbidity and mortality worldwide. The virulence and pathogenic nature of influenza virus strains vary greatly, and is often determined by the constellation of viral genes. While seasonal influenza viruses cause a mild upper respiratory infection in healthy individuals, highly pathogenic avian influenza viruses (HPAIV) of the H5N1 subtype replicate in the lower respiratory tract, causing fatal viral pneumonia characterized by pulmonary edema and vascular leakage. In addition, H5N1 infections are associated with an uncontrolled activation of host immune responses, which contributes to the severity of disease. The role of viral and host factors in the enhanced virulence and pulmonary immunopathology characteristic of HPAIV infections remain unknown. To this end, we engineered H5N1 viruses with restricted tropism through the incorporation of microRNA (miRNA) target sites into the viral genome. Specifically, we generated an H5N1 virus carrying endothelial cell specific miR-126 target sites (H5N1-126T), such that viral replication was abrogated in endothelial cells without impeding replication in other cell types. In our recent studies, H5N1-126T infected mice and ferrets showed significantly reduced virulence and vascular leakage in the lungs as compared to the control H5N1-ScrbT virus infected group, despite similar viral loads in the lungs of mice and nasal washes of ferrets. To our knowledge, this is the first study to demonstrate the importance of endothelial cell tropism to H5N1 pathogenesis. The current proposal will (1) determine the consequence of endothelial cell infection to barrier integrity and function, (2) investigate the role of endothelial cell tropism in orchestrating immune responses, and (3) assess the contribution of viral tropism to the pathogenesis of other highly virulent strains including H7N7, 1918 H1N1, and H7N9 in mice and ferrets. The knowledge gained from these studies will help to elucidate the mechanism by which virulent influenza viruses cause severe disease and aid in the development of novel therapeutic strategies against virulent influenza strains. Furthermore, these studies will reveal how viral tropism modulates host antiviral responses and will pave the way for the rational design of novel live attenuated vaccines with restricted tropism.

抽象的 流感病毒对人类健康构成严重威胁，引起季节性流行病和偶尔 大流行病导致全球发病率和死亡率很大。毒力和致病性 流感病毒菌株变化很大，通常由病毒基因的星座确定。季节性 流感病毒在健康个体中引起轻度的上呼吸道感染，高度致病的鸟类 H5N1亚型在下呼吸道中复制的流感病毒（HPAIV），导致致命病毒 肺炎的特征是肺水肿和血管渗漏。另外，H5N1感染是 与宿主免疫反应的不受控制的激活相关，这有助于 疾病。病毒和宿主因素在增强的毒力和肺部免疫病理学中的作用 HPAIV感染的特征仍然未知。为此，我们对H5N1病毒进行了限制 通过将microRNA（miRNA）靶位点掺入病毒基因组中的向热力主义。具体来说，我们 产生了携带内皮细胞特异性miR-126靶位点（H5N1-126T）的H5N1病毒，以便病毒 在内皮细胞中废除了复制，而不会阻碍其他细胞类型的复制。在我们最近的 研究，H5N1-126T感染的小鼠和雪貂在毒力和血管泄漏显着降低 与对照H5N1-SCRBT病毒感染组相比，肺部尽管肺部有相似的病毒负荷 雪貂的小鼠和鼻腔洗涤。据我们所知，这是第一项证明重要性的研究 H5N1发病机理的内皮细胞向量。当前的提案将（1）确定 内皮细胞感染对屏障完整性和功能，（2）研究内皮细胞偏向主义在 策划免疫反应，（3）评估病毒tropism对其他发病机理的贡献 小鼠和雪貂中的高毒素包括H7N7，1918 H1N1和H7N9。从中获得的知识 这些研究将有助于阐明毒性流感病毒引起严重疾病的机制 并有助于开发针对有毒流感菌株的新型治疗策略。此外， 这些研究将揭示病毒性向往往如何调节宿主抗病毒反应，并为 新颖的活衰减疫苗的理性设计与恐怖症。

项目成果

Genome-wide CRISPR/Cas9 Screen Identifies Host Factors Essential for Influenza Virus Replication.

• DOI: 10.1016/j.celrep.2018.03.045
• 发表时间: 2018-04-10
• 期刊: Cell reports
• 影响因子: 8.8
• 作者: Han J;Perez JT;Chen C;Li Y;Benitez A;Kandasamy M;Lee Y;Andrade J;tenOever B;Manicassamy B
• 通讯作者: Manicassamy B

Broadly Protective Strategies Against Influenza Viruses: Universal Vaccines and Therapeutics.

针对流感病毒的广泛保护策略：通用疫苗和治疗方法。

• DOI: 10.3389/fmicb.2020.00135
• 发表时间: 2020
• 期刊: Frontiers in microbiology
• 影响因子: 5.2
• 作者: Vogel,OliviaA;Manicassamy,Balaji
• 通讯作者: Manicassamy,Balaji

Development of a Single-Cycle Infectious SARS-CoV-2 Virus Replicon Particle System for Use in Biosafety Level 2 Laboratories.

• DOI: 10.1128/jvi.01837-21
• 发表时间: 2022-02-09
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Malicoat J;Manivasagam S;Zuñiga S;Sola I;McCabe D;Rong L;Perlman S;Enjuanes L;Manicassamy B
• 通讯作者: Manicassamy B

Endothelial cell tropism is a determinant of H5N1 pathogenesis in mammalian species.

• DOI: 10.1371/journal.ppat.1006270
• 发表时间: 2017-03
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Tundup S;Kandasamy M;Perez JT;Mena N;Steel J;Nagy T;Albrecht RA;Manicassamy B
• 通讯作者: Manicassamy B