Identifying Poxvirus Receptors

识别痘病毒受体

• 批准号: 9809258
• 负责人: BRIAN M WARD
• 金额: $ 21.75万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9809258
• 关键词: Antigens; Antiviral Agents; Binding; Binding Proteins; Biology; Cell Separation; Cell membrane; Cell surface; Cells; Complementary DNA; Complex; Conflict (Psychology); DNA Viruses; Data; Development; Disease; Ectromelia; Eukaryotic Cell; Exhibits; Family; Family member; Flow Cytometry; Gene Expression; Genes; Goals; Human; Human Genome; Individual; Intelligence; Knowledge; Libraries; Microscopy; Modality; Modeling; Molecular; Monkeypox; Mouse Pox Virus; Myxoma; Myxoma virus; Oncolytic; Oncolytic viruses; Orthopoxvirus; Pathogenesis; Plasmids; Poxviridae; Process; Proteins; Protocols documentation; Public Health; Recombinants; Recording of previous events; Reporting; Research; Severity of illness; Smallpox; Surface; System; Testing; Time; Tropism; Vaccines; Vaccinia; Vaccinia virus; Validation; Viral; Viral Pathogenesis; Viral Proteins; Virion; Virus; Virus Diseases; Virus Receptors; Work; Yeast Model System; Yeasts; base; cDNA Library; cell type; deep sequencing; design; follow-up; genetic manipulation; insight; member; prevent; receptor; response; screening; success; vector; virus tropism

Poxviruses are a large family of DNA viruses capable of infecting and causing disease in humans. Its members include variola, the causative agent of smallpox, and monkeypox, which causes a smallpox-like disease that can be lethal. In addition to these public health issues, poxviruses are being developed and used as vaccine platforms and oncolytic agents. The prototypical poxvirus, vaccinia, is able to bind and enter, every cell type tested. Currently, an unambiguous cellular receptor has not been identified for poxviruses and represents a major gap in our knowledge of their lifecycle. The long-term goal of this project is to determine which cellular receptor(s) are capable of interacting with which of the ~30 viral proteins found on the surface of poxvirus virions so that pseudotyping and cell targeting can be achieved. Our studies will also help in understanding the vast tropism exhibited by vaccinia virus and other members of this group. The immediate goal is to develop a system to screen for cellular proteins that interact with vaccinia virus. These results will then be expanded to other members of this family (ectromelia virus and myxoma virus) to determine if they utilize the same receptors or if they have evolved to use different receptors. Our specific aims are; 1) Screen a cDNA library using yeast surface display for cellular proteins that the intracellular mature form of vaccinia virus can bind. 2) Prioritize and validate hits for vaccinia virus. Determine if ectromelia, and myxoma can bind putative receptors identified in the screen. The results obtained will identify cellular receptor(s) used by poxviruses and provide greater insight into the molecular mechanism of the broad tropism of these viruses, along with establishing a new receptor screening modality for viruses that have a broad tropism.

痘病毒是一大类 DNA 病毒，能够感染人类并引起人类疾病。其成员包括天花（天花的病原体）和猴痘（引起类似天花的致命疾病）。除了这些公共卫生问题之外，痘病毒还被开发并用作疫苗平台和溶瘤剂。典型的痘病毒痘苗病毒能够结合并进入每种测试的细胞类型。目前，尚未确定痘病毒的明确细胞受体，这代表了我们对其生命周期知识的重大空白。该项目的长期目标是确定哪些细胞受体能够与痘病毒病毒粒子表面发现的约 30 种病毒蛋白中的哪些相互作用，从而实现假型分析和细胞靶向。我们的研究还将有助于了解痘苗病毒和该组其他成员所表现出的巨大趋向性。近期目标是开发一种系统来筛选与痘苗病毒相互作用的细胞蛋白。然后，这些结果将扩展到该家族的其他成员（节肢动物病毒和粘液瘤病毒），以确定它们是否利用相同的受体，或者是否已经进化到使用不同的受体。我们的具体目标是； 1) 使用酵母表面展示筛选 cDNA 文库，寻找细胞内成熟形式的痘苗病毒可以结合的细胞蛋白。 2) 优先考虑并验证痘苗病毒的命中。确定异常肢和粘液瘤是否可以结合屏幕中识别的推定受体。获得的结果将鉴定痘病毒使用的细胞受体，并为这些病毒的广泛向性的分子机制提供更深入的了解，同时为具有广泛向性的病毒建立新的受体筛选模式。