Defining viral-host interactions between arthritogenic alphaviruses and MARCO

定义致关节炎甲病毒和 MARCO 之间的病毒-宿主相互作用

• 批准号: 10633062
• 负责人: Frances Shieh Li
• 金额: $ 3.68万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10633062
• 关键词: Affect; Alphavirus; Amino Acids; Arboviruses; Arthritogenic; Binding; Binding Sites; Biochemical; Biological Assay; Blood; Cells; Charge; Chikungunya virus; Circulation; Clinical; Communicable Diseases; Culicidae; Disease Outbreaks; Evaluation; Excision; Foundations; Genetic Polymorphism; Geography; Glutamates; Glycoproteins; Human; In Vitro; Individual; Infection; Innate Immune System; Invertebrates; Kinetics; Knowledge; Kupffer Cells; Laboratories; Life Cycle Stages; Ligands; Low-Density Lipoproteins; Lysine; Mass Spectrum Analysis; Methylation; Modification; Molecular; Mus; Mutate; Mutation; Outcome; Pathogenesis; Pattern recognition receptor; Periodicals; Population; Positioning Attribute; Post-Translational Protein Processing; Predisposition; Proteins; Public Health; Publishing; RXR; Reader; Reporting; Role; SR-A proteins; SRCR proteins; Scavenger Receptor Cysteine-Rich Domain; Serum; Severity of illness; Site; Surface; Surface Plasmon Resonance; Tuberculosis; Variant; Vertebrates; Viral; Viremia; Virion; Virus; acetyl-LDL; chikungunya infection; feeding; in vivo; insight; nonhuman primate; particle; pathogen; scavenger receptor; targeted treatment; transmission process; virus host interaction

PROJECT SUMMARY Arboviruses maintained in a human-mosquito-human transmission cycle are responsible for fueling periodic outbreaks worldwide and are an increasing public health threat. A critical feature of arbovirus transmission cycles, and a major determinant of their geographic spread and pathogenesis, is the magnitude and duration of viremia in vertebrate hosts. However, few studies have investigated the molecular determinants of viremia. Recent studies published by the Morrison laboratory demonstrated that the murine scavenger receptor MARCO on liver macrophages removes chikungunya (CHIKV) particles and other arthritogenic alphaviruses, including Ross River (RRV) and o’nyong ‘nyong (ONNV) viruses, from murine circulation due to recognition of the lysine (K) residue at position 200 of CHIKV and ONNV E2 glycoprotein and 251 of RRV E2 glycoprotein. My preliminary studies further revealed that CHIKV clearance is also abrogated when mutations were introduced at glutamate (E)208 of E2 and K61 of E1 glycoproteins, and mass spectrometry analysis of the biochemical features important for viral clearance suggested that E1 K61 is methylated. Further analysis of position 208 of CHIKV E2 glycoprotein revealed the importance of a negative charge at this position for CHIKV removal from circulation. As a pattern recognition receptor, MARCO recognizes modified self and non-self molecules, and polymorphisms in human MARCO can predispose carriers to infectious diseases such as tuberculosis. Because the scavenger receptor cysteine-rich (SRCR) domain of MARCO is a binding site for endogenous ligands, such as modified low-density lipoprotein, I hypothesize that the SRCR domain of MARCO stably and noncovalently interacts with an exposed interface between the E1 and E2 glycoproteins of CHIKV, allowing for the removal of viral particles from circulation and a reduction in both the magnitude and duration of viremia. In Aim 1, I will define the residues and biochemical features of CHIKV important for MARCO-dependent clearance from circulation by manipulating surface features of virus particles, assessing how specific mutations impact viral dissemination, and identifying post-translational modifications at specific sites in the E1 and E2 glycoproteins. In Aim 2, I will elucidate the sites on MARCO responsible for binding arthritogenic alphaviruses with cell-based and biochemical approaches. In addition, I will determine the extent to which virus particles interact with human MARCO, and whether known polymorphisms in MARCO affect virus-MARCO interactions, viremia, or clinical outcomes. Taken together, by defining the molecular mechanism of interaction between MARCO and CHIKV, this proposal could provide insights into factors that influence alphaviral pathogenesis, elucidate the relationship between MARCO polymorphisms and viremia, and identify individuals or populations with an increased susceptibility to severe alphaviral infections and outbreaks, respectively.

项目摘要 维持在人类 - 摩斯高族人 - 人类传输周期中的arbovirus是为了定期加油 全球爆发，是日益增长的公共卫生威胁。 Arbovirus传输周期的关键特征， 其地理扩散和发病机理的主要决定因素是病毒血症的幅度和持续时间 在脊椎动物主机中。但是，很少有研究研究了病毒血症的分子决定剂。最近的 莫里森实验室（Morrison Laboratory）发表的研究表明，肝脏的鼠清道夫接收器Marco在肝脏上 巨噬细胞去除chikungunya（chikv）颗粒和其他关节型α病毒，包括罗斯 河流（RRV）和O'Nyong'Nyong（ONNV）病毒，由于赖赛（K）的识别而引起的鼠循环。 CHIKV和ONNV E2糖蛋白的200位以及RRV E2糖蛋白的251位残留物。我的初步 研究进一步表明，当在谷氨酸引入突变时，CHIKV清除也被废除 （E）E2糖蛋白的E2和K61的208，生化特征的质谱分析很重要 对于病毒清除，表明E1 K61是甲基化的。对Chikv E2的位置208的进一步分析 糖蛋白揭示了该位置上的负电荷对从循环中去除的重要性的重要性。 作为模式识别受体，Marco识别经过修改的自我和非自我分子以及多态性 在人类中，Marco可以使携带者易于传染病，例如结核病。因为清道夫 Marco的受体半胱氨酸（SRCR）结构域是内源配体的结合位点，例如修饰 低密度脂蛋白，我假设Marco的SRCR结构域稳定且无创相互作用 CHIKV的E1和E2糖蛋白之间的裸露接口，允许去除 病毒颗粒的循环颗粒和病毒血症的幅度和持续时间的减少。在AIM 1中， 我将定义Chikv的残差和生化特征，对于Marco依赖的清除很重要 通过操纵病毒颗粒的表面特征循环，评估特定突变如何影响病毒 在E1和E2糖蛋白中的特定部位进行传播，并鉴定翻译后修饰。在 AIM 2，我将阐明Marco上负责与基于细胞和基于细胞和基于细胞的关节性α病毒结合的地点 生化方法。另外，我将确定病毒颗粒与人相互作用的程度 Marco，以及Marco中已知的多态性是否影响病毒 - 玛科相互作用，病毒血症或临床 结果。通过定义Marco和Chikv之间相互作用的分子机制一起综合起来， 该提案可以提供对影响字母发病机理的因素，阐明关系的因素 在Marco多态性和病毒血症之间，并识别出增加的个体或人群 分别对严重的字母感染和暴发的敏感性。