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.

项目概要 维持在人类-蚊子-人类传播周期中的虫媒病毒负责促进周期性传播 疫情在全球范围内暴发，并成为日益严重的公共卫生威胁。虫媒病毒传播周期的一个关键特征， 其地理传播和发病机制的一个主要决定因素是病毒血症的程度和持续时间 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。最近的 Morrison 实验室发表的研究表明，小鼠肝脏中的清道夫受体 MARCO 巨噬细胞清除基孔肯雅病毒 (CHIKV) 颗粒和其他导致关节炎的甲病毒，包括罗斯病毒 River (RRV) 和 o’nyong ‘nyong (ONNV) 病毒，由于识别赖氨酸 (K) 而来自小鼠循环 CHIKV 和 ONNV E2 糖蛋白的 200 位残基以及 RRV E2 糖蛋白的 251 位残基。我的初步 研究进一步表明，当谷氨酸引入突变时，CHIKV 清除也会被取消 (E)E2的208和E1糖蛋白的K61，以及重要生化特征的质谱分析 病毒清除表明 E1 K61 被甲基化。 CHIKV E2 208位点的进一步分析 糖蛋白揭示了该位置负电荷对于 CHIKV 从循环中去除的重要性。 作为模式识别受体，MARCO 识别修饰的自身和非自身分子以及多态性 在人类中，MARCO 会使携带者易患结核病等传染病。因为拾荒者 MARCO 的受体富含半胱氨酸 (SRCR) 结构域是内源性配体的结合位点，例如修饰的配体 低密度脂蛋白，我假设 MARCO 的 SRCR 结构域稳定且非共价地相互作用 CHIKV 的 E1 和 E2 糖蛋白之间有暴露的界面，可以去除 病毒颗粒从循环中排出，并减少病毒血症的程度和持续时间。在目标 1 中， 我将定义对 MARCO 依赖性清除很重要的 CHIKV 残留物和生化特征 通过操纵病毒颗粒的表面特征来评估特定突变如何影响病毒的循环 传播，并识别 E1 和 E2 糖蛋白特定位点的翻译后修饰。在 目标 2，我将阐明 MARCO 上负责将致关节炎 α 病毒与基于细胞和 生化方法。此外，我还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。综上所述，通过定义 MARCO 和 CHIKV 之间相互作用的分子机制， 该提议可以深入了解影响甲病毒发病机制的因素，阐明其中的关系 MARCO 多态性和病毒血症之间的关系，并确定病毒血症增加的个体或群体 分别对严重甲病毒感染和爆发的易感性。