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A New Targeting Approach to Inhibit Budding of the Ebola Virus

抑制埃博拉病毒萌芽的新靶向方法

基本信息

批准号：
9763445
负责人：
Robert Virgil Stahelin
金额：
$ 22.64万
依托单位：
PURDUE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-14 至 2021-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9763445
关键词：
Affinity Amino Acid Sequence Amino Acids Animal Model Animals Antibodies Antibody Therapy Biological Assay Biological Models Biophysics Cell Survival Cell membrane Cells Chemicals Clinical Communicable Diseases Computer Analysis Data Dimerization Disease Outbreaks Drug Targeting Ebola virus Equilibrium Escape Mutant FDA approved Family Fatality rate Filoviridae Infections Filovirus Frankfurt-Marburg Syndrome Virus Fright Generations Genes Glycoproteins Human In Vitro Laboratories Lead Life Cycle Stages Lipid Bilayers Lipid Binding Lipids Malignant Neoplasms Mammalian Cell Mediating Methods Modeling Mutate Mutation N-terminal Patients Penetration Peptides Pharmacology Plasma Cells Preventive measure Process Production Property Proteins Public Health Recording of previous events Structural Protein Structure Testing Therapeutic Vaccine Clinical Trial Vaccine Therapy Vaccines Viral Viral Matrix Proteins Virion Virus Virus Assembly Virus Replication Virus-like particle Western Africa alpha helix base chemical synthesis cost design dimer experimental study insight lead candidate monomer pandemic disease protein protein interaction protein structure small molecule therapeutic target therapeutic vaccine tool virus envelope

项目摘要

Abstract: Lipid-enveloped viruses replicate and bud from host cell membranes where they acquire their lipid coat. Understanding the budding processes of several viruses has had significant impact on elucidating the viral life cycle and identifying therapeutic targets. Filoviruses have a filamentous lipid- envelope and despite being discovered more than 30 years ago, not much is known on how they assemble and bud from the host cell plasma membrane. Filoviruses, which include Ebola virus (EBOV), have a high fatality rate and there is still a lack of FDA approved therapeutics or vaccines for treatment. Moreover, the EBOV glycoprotein, the prime target of antibody and vaccine therapy undergoes a high rate of mutation in animal and human studies and escape mutant of glycoprotein have been found as EBOV is passaged through animal models. Filoviruses encode seven genes including the viral matrix protein VP40, which regulates budding from the host cell. VP40 as the only filovirus protein expressed in mammalian cells is sufficient to produce virus like particles (VLPs) nearly indistinguishable from live virions. Thus, VP40 has served as a model to study viral budding outside of BSL-4 laboratories. VP40 has been shown to be a dimer, which is mediated by a-helical interactions in its N-terminal domain (NTD). Mutation of residues in the NTD of VP40 that mediate dimerization is sufficient to abrogate viral budding in model systems. To date, little is known about how VP40 monomer/dimer equilibrium and biophysics of oligomer assembly are regulated as well as if VP40 is a viable drug target in the viral life cycle. The central hypothesis of this R21 proposal is that generation of a new chemical toolkit based upon stapled a-helical peptides can be used to study VP40 assembly and inhibit VP40 dimerization. In specific aim 1, we will design and synthesize lead candidate stapled a-helical peptides that target the VP40 dimer interface. We will elucidate the optimal amino acid sequences and chemical linker of stapled a-helical peptides using computational analysis. We hypothesize that optimization of the stapled helices can be performed to block VP40 dimer formation in vitro and in cells. We will use computational analysis and a rapid chemical synthesis method to generate lead candidates for quantitative analysis. Specific aim 2 will investigate the mechanism by which stapled a- helical peptides interact with VP40 and inhibit VP40 dimerization and budding of VLPs. Quantitative assays of VP40 dimer formation, VP40 lipid-binding, and budding of VLPs will be assessed to decipher the ability of lead compounds to inhibit dimer formation and subsequent budding. Taken together, these studies should produce new and important mechanistic insight into the viability of VP40 as a drug target and a better biophysical understanding of the properties that govern VP40 assembly.

脂质包膜病毒在宿主细胞膜上进行复制和芽化，并在宿主细胞膜上获得它们的生存能力