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: 10386284
• 负责人: Frances Shieh Li
• 金额: $ 3.54万
• 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/10386284
• 关键词: Affect; Alphavirus; Amino Acids; Arboviruses; Arthritogenic; Binding; Binding Sites; Biochemical; Biological Assay; Blood; Blood Circulation; Cells; Charge; Chikungunya virus; 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; O' nyong-nyong virus; Outcome; Pathogenesis; Pattern recognition receptor; Periodicity; Population; Positioning Attribute; Post-Translational Protein Processing; Predisposition; Proteins; Public Health; Publishing; RXR; Reader; Reporting; Rivers; 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; base; chikungunya; 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-蚊-人传播周期中的虫媒病毒负责助长周期性的 疫情在全球范围内爆发，对公共卫生构成越来越大的威胁。虫媒病毒传播周期的一个关键特征， 病毒血症的程度和持续时间是其地理分布和发病机制的主要决定因素 在脊椎动物宿主中。然而，很少有研究调查病毒血症的分子决定因素。最近 莫里森实验室发表的研究表明， 巨噬细胞清除基孔肯雅病毒（CHIKV）颗粒和其他致关节炎的甲病毒，包括罗斯病毒（Ross River（RRV）和o 'nyong'nyong（ONNV）病毒，由于识别赖氨酸（K）而来自鼠循环 在CHIKV和ONNV E2糖蛋白的位置200和RRV E2糖蛋白的位置251处的残基。我的初步 研究进一步揭示，当在谷氨酸处引入突变时，CHIKV清除也被废除， （E）208的E2和K61的E1糖蛋白，以及质谱分析的重要生化特征 病毒清除表明E1 K61是甲基化的。CHIKV E2的位置208的进一步分析 糖蛋白揭示了在该位置处的负电荷对于CHIKV从循环中去除的重要性。 作为一种模式识别受体，MARCO识别修饰的自我和非自我分子，以及多态性 在人类中，MARCO可使携带者易患传染病，如肺结核。因为食腐动物 MARCO的受体富含半胱氨酸（SRCR）结构域是内源性配体（如修饰的配体）的结合位点， 低密度脂蛋白，我假设MARCO的SRCR结构域稳定和非共价相互作用 在CHIKV的E1和E2糖蛋白之间具有暴露的界面，允许去除CHIKV的E1和E2糖蛋白。 从循环中清除病毒颗粒，并减少病毒血症的程度和持续时间。在目标1中， 我将定义CHIKV的残基和生化特征，这些残基和生化特征对于MARCO依赖性清除从 通过操纵病毒颗粒的表面特征，评估特定突变如何影响病毒 传播，并鉴定在E1和E2糖蛋白的特定位点的翻译后修饰。在 目的2，我将阐明MARCO上负责结合致关节炎甲病毒的位点， 生物化学方法。此外，我将确定病毒颗粒与人类相互作用的程度， MARCO，以及MARCO中已知的多态性是否影响病毒-MARCO相互作用、病毒血症或临床 结果。总之，通过定义MARCO和CHIKV之间相互作用的分子机制， 这一建议可以提供对影响甲病毒发病机制的因素的见解，阐明甲病毒与病毒之间的关系， MARCO多态性和病毒血症之间的关系，并确定具有增加的 严重甲病毒感染和爆发的易感性。