Host-virus interactions in the control of filovirus entry

控制丝状病毒进入的宿主-病毒相互作用

• 批准号: 10061541
• 负责人: Kartik Chandran
• 金额: $ 47.53万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-05 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10061541
• 关键词: Africa; Antiviral Agents; Antiviral Therapy; Binding; Biogenesis; Biological Assay; Biology; Bioterrorism; Caspase; Cathepsins; Cell membrane; Cells; Cellular biology; Chemicals; Cleaved cell; Collaborations; Complement; Complex; Coupled; Cysteine; Cytoplasm; Data; Development; Disease; Disease Outbreaks; Dyes; Ebola virus; Endosomes; Filoviridae Infections; Filovirus; Genetic; Genetic Screening; Glycoproteins; Goals; Grant; Haploidy; Human; Image; In Vitro; Infection; Integration Host Factors; Invaded; Light; Lipid Bilayers; Lipids; Lysosomes; Mediating; Membrane; Membrane Fusion; Molecular; Molecular Conformation; Nucleocapsid; Pathway interactions; Peptide Hydrolases; Play; Principal Investigator; Process; Protease Inhibitor; Proteins; Proteomics; Reaction; Reagent; Resources; Role; Secondary to; Site; Structure; Testing; Therapeutic; Time; Vaccines; Viral; Viral Hemorrhagic Fevers; Virion; Virus; Virus Diseases; Virus-like particle; Work; Zoonoses; base; cholesterol transporters; comparative; design; disease transmission; experimental study; genetic approach; in vitro Assay; in vitro Model; late endosome; member; multidisciplinary; mutant; novel; particle; prevent; programs; receptor; reconstitution; tool; trafficking; virology; virus host interaction

The unprecedented 2014–2015 Ebola virus (EBOV) outbreak in West Africa has highlighted the urgent, unfulfilled need for vaccines and therapeutics to prevent and treat infections by EBOV and other filoviruses. However, their development is challenged by crucial gaps in our understanding of the virus-host molecular interactions that underpin filovirus infection, disease, and transmission. Recent work on the entry mechanism of filoviruses has revealed that these agents are prototypic members of a group of `late-penetrating' viruses that extensively exploit host endo/lysosomal factors and pathways to invade cells. During the previous grant period, we found that host endosomal cysteine proteases (cysteine cathepsins) program the viral glycoprotein GP to bind the endosomal cholesterol transporter Niemann-Pick C1 (NPC1), which we showed to be an essential intracellular entry receptor. Despite these and other discoveries, however, the culminating steps in filovirus entry—GP-mediated fusion between viral and cellular lipid bilayers, and cytoplasmic escape of the viral nucleocapsid core—remain enigmatic. The overarching goals of this renewal application are to define the molecular mechanism of filovirus membrane fusion, and to identify new filovirus entry host factors with antiviral potential. To fulfill these objectives, the principal investigator has assembled a multidisciplinary team that includes a field leader in the chemical biology of proteases, a leading expert on the biogenesis and functions of endosomes and lysosomes, and a virologist who will extend findings with surrogate viruses to authentic filoviruses. Together we will: (1) Define viral and cellular requirements for filovirus membrane fusion, dissect steps in the fusion reaction, and identify the elusive filovirus fusion trigger factor(s). (2) Determine the roles of newly- identified host factors in viral trafficking to membrane fusion sites, and employ proteomic approaches to uncover novel host factors involved in filovirus membrane fusion.

2014-2015年西非前所未有的埃博拉病毒（EBOV）爆发凸显了对预防和治疗EBOV和其他丝状病毒感染的疫苗和治疗剂的迫切而未满足的需求。然而，它们的发展受到我们对支持丝状病毒感染，疾病和传播的病毒-宿主分子相互作用的理解的关键差距的挑战。丝状病毒进入机制的最新研究表明，这些药物是一组“晚期穿透”病毒的原型成员，这些病毒广泛利用宿主内/溶酶体因子和途径侵入细胞。在之前的资助期间，我们发现宿主内体半胱氨酸蛋白酶（半胱氨酸组织蛋白酶）对病毒糖蛋白GP进行编程，以结合内体胆固醇转运蛋白尼曼-匹克C1（NPC 1），我们证明NPC 1是一种必需的细胞内进入受体。然而，尽管有这些和其他发现，丝状病毒进入GP介导的病毒和细胞脂质双层之间的融合以及病毒核衣壳核心的细胞质逃逸的最终步骤仍然是谜。本次更新申请的总体目标是确定丝状病毒膜融合的分子机制，并鉴定具有抗病毒潜力的新丝状病毒进入宿主因子。为了实现这些目标，主要研究者组建了一个多学科团队，其中包括蛋白酶化学生物学领域的领导者，核内体和溶酶体的生物发生和功能方面的领先专家，以及将替代病毒的发现扩展到真实丝状病毒的病毒学家。我们将共同：（1）定义丝状病毒膜融合的病毒和细胞要求，剖析融合反应中的步骤，并鉴定难以捉摸的丝状病毒融合触发因子。(2)确定新鉴定的宿主因子在病毒运输至膜融合位点中的作用，并采用蛋白质组学方法来发现参与丝状病毒膜融合的新型宿主因子。

项目成果

A Glycoprotein Mutation That Emerged during the 2013-2016 Ebola Virus Epidemic Alters Proteolysis and Accelerates Membrane Fusion.

• DOI: 10.1128/mbio.03616-20
• 发表时间: 2021-02-16
• 期刊: mBio
• 影响因子: 6.4
• 作者: Fels JM;Bortz RH 3rd;Alkutkar T;Mittler E;Jangra RK;Spence JS;Chandran K
• 通讯作者: Chandran K

Structural basis of broad ebolavirus neutralization by a human survivor antibody.

人类幸存者抗体广泛中和埃博拉病毒的结构基础。

• DOI: 10.1038/s41594-019-0191-4
• 发表时间: 2019
• 期刊: Nature structural & molecular biology
• 影响因子: 16.8
• 作者: West,BrandynR;Wec,AnnaZ;Moyer,CrystalL;Fusco,MarnieL;Ilinykh,PhilippA;Huang,Kai;Wirchnianski,ArielS;James,RebekahM;Herbert,AndrewS;Hui,Sean;Goodwin,Eileen;Howell,KatieA;Kailasan,Shweta;Aman,MJavad;Walker,LauraM;Dye
• 通讯作者: Dye

Two Distinct Lysosomal Targeting Strategies Afford Trojan Horse Antibodies With Pan-Filovirus Activity.

• DOI: 10.3389/fimmu.2021.729851
• 发表时间: 2021
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Wirchnianski AS;Wec AZ;Nyakatura EK;Herbert AS;Slough MM;Kuehne AI;Mittler E;Jangra RK;Teruya J;Dye JM;Lai JR;Chandran K
• 通讯作者: Chandran K

Systematic Analysis of Monoclonal Antibodies against Ebola Virus GP Defines Features that Contribute to Protection.

• DOI: 10.1016/j.cell.2018.07.033
• 发表时间: 2018-08-09
• 期刊: Cell
• 影响因子: 64.5
• 作者: Saphire EO;Schendel SL;Fusco ML;Gangavarapu K;Gunn BM;Wec AZ;Halfmann PJ;Brannan JM;Herbert AS;Qiu X;Wagh K;He S;Giorgi EE;Theiler J;Pommert KBJ;Krause TB;Turner HL;Murin CD;Pallesen J;Davidson E;Ahmed R;Aman MJ;Bukreyev A;Burton DR;Crowe JE Jr;Davis CW;Georgiou G;Krammer F;Kyratsous CA;Lai JR;Nykiforuk C;Pauly MH;Rijal P;Takada A;Townsend AR;Volchkov V;Walker LM;Wang CI;Zeitlin L;Doranz BJ;Ward AB;Korber B;Kobinger GP;Andersen KG;Kawaoka Y;Alter G;Chandran K;Dye JM;Viral Hemorrhagic Fever Immunotherapeutic Consortium
• 通讯作者: Viral Hemorrhagic Fever Immunotherapeutic Consortium

S-Palmitoylation and Sterol Interactions Mediate Antiviral Specificity of IFITMs.

• DOI: 10.1021/acschembio.2c00176
• 发表时间: 2022-08-19
• 期刊: ACS CHEMICAL BIOLOGY
• 影响因子: 4
• 作者: Das, Tandrila;Yang, Xinglin;Lee, Hwayoung;Garst, Emma H.;Valencia, Estefania;Chandran, Kartik;Im, Wonpil;Hang, Howard C.
• 通讯作者: Hang, Howard C.