Alpha-v Integrins and Adenovirus Cell Entry

Alpha-v 整合素和腺病毒细胞进入

• 批准号: 8686919
• 负责人: Glen R Nemerow
• 金额: $ 46.43万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-01-20 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8686919
• 关键词: Adenovirus Infections; Adenovirus Protein; Adenovirus Vector; Adenoviruses; Atomic Force Microscopy; Binding; Biology; Capsid; Capsid Proteins; Cardiovascular Diseases; Cell membrane; Cells; Cellular Structures; Clinical Treatment; Clinical Trials; Collaborations; Complex; Cryoelectron Microscopy; Defensins; Development; Endosomes; Event; Fiber; Foundations; Funding; Genome; Goals; Human Adenoviruses; Image; Immune; Immune system; Integration Host Factors; Integrins; Investigation; Kinetics; Knowledge; Laboratories; Lead; Life Cycle Stages; Light; Location; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Mechanics; Mediating; Membrane; Membrane Lipids; Minor; Modeling; Molecular; Molecular Genetics; Molecular Virology; Mutagenesis; Mutation; NMR Spectroscopy; Pathway interactions; Penetration; Phase; Property; Proteins; Research Institute; Research Personnel; Resolution; Role; Solutions; Structure; Surface; Surface Plasmon Resonance; Tensile Strength; Vaccines; Viral; Viral Vector; Virion; Virus; X ray diffraction analysis; X-Ray Diffraction; anti-viral gene delivery; biophysical properties; biophysical techniques; experience; gene therapy; improved; insight; multidisciplinary; nanoindentation; particle; penton base; research study; solid state nuclear magnetic resonance; stoichiometry; structural biology; thermostability

DESCRIPTION (provided by applicant): While adenoviral vectors continue to be employed in phase 1-III clinical trials for the treatment of cancers and cardiovascular diseases, we still have an incomplete understanding of the virus entry pathway. During the previous funding period, researchers in the Nemerow and Reddy laboratories at the Scripps Research Institute solved the crystal structure of adenovirus at near atomic resolution thereby revealing the locations, folds and interactions of several viral capsid proteins. Additional investigations in the Nemerow and Stewart laboratories (Vanderbilt) discovered that specific host factors either destabilize (alpha v integrins) the virus capsid or restrict (defensins) capsid disassembly. Although this information has shed further light on the mechanisms involved in virus entry into cells, numerous gaps remain in our knowledge of virus disassembly and membrane penetration events. In this proposal, a team of experienced investigators with expertise in molecular virology, structural biology, and biophysical techniques will investigate the precise mechanisms involved in capsid disassembly and membrane penetration by protein VI. X-ray diffraction and cryoEM structural analyses of several key capsid proteins including proteins VI, V and hexon will be undertaken. Advanced imaging of adenovirus by atomic force microscopy will be employed to measure the tensile strength and mechanical properties of the viral capsid. Native and denatured mass spectrometry will be used to determine the copy number of protein VI in the virion as well as its association states with other capsid proteins. Protein VI mutagenesis studies will analyze the role of protein VI in capsid assembly/disassembly and endosome penetration. The long-term goal of these studies is to obtain a more complete picture of host cell invasion by adenovirus. Such knowledge should bolster the emerging knowledge of how non-enveloped viruses breach host cell membranes as well as provide a foundation for the development of safer and more efficient viral vectors.

描述（由申请人提供）：虽然腺病毒载体继续在第1阶段III临床试验中用于治疗癌症和心血管疾病，但我们仍然有 对病毒进入途径的不完全理解。在上一个资金期间，Scripps研究所的Nemerow和Reddy实验室的研究人员在近原子分辨率下解决了腺病毒的晶体结构，从而揭示了几种病毒式衣壳蛋白的位置，折叠和相互作用。 Nemerow和Stewart Laboratories（Vanderbilt）的其他调查发现，特定的宿主因素要么破坏病毒capsid或限制（Defensins）CapsId拆卸。尽管这些信息进一步阐明了病毒进入细胞的机制，但我们对病毒拆卸和膜穿透事件的了解仍然存在许多差距。在该提案中，由经验丰富的研究人员团队在分子病毒学，结构生物学和生物物理技术方面具有专业知识，将研究蛋白质VI涉及的胶囊拆卸和膜渗透所涉及的确切机制。将进行几种包括蛋白质VI，V和Hexon在内的几种关键衣壳蛋白的X射线衍射和冷冻结构分析。通过原子力显微镜对腺病毒的高级成像将用于测量病毒衣壳的拉伸强度和机械性能。天然和变性的质谱法将用于确定病毒体中蛋白质VI的拷贝数及其与其他衣壳蛋白的缔合状态。蛋白质VI诱变研究将分析蛋白质VI在衣壳组装/拆卸和内体穿透性中的作用。这些研究的长期目标是获得腺病毒宿主细胞侵袭的更完整的图像。这种知识应该增强有关非发育病毒如何违反宿主细胞膜的新兴知识，并为开发更安全，更有效的病毒载体的发展提供了基础。