Determining mechanisms of innate immune modulation by ADP-ribosylation

通过 ADP-核糖基化确定先天免疫调节机制

基本信息

批准号：
10027966
负责人：
Anthony R Fehr
金额：
$ 36.1万
依托单位：
UNIVERSITY OF KANSAS LAWRENCE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-15 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10027966
关键词：
ADP Ribose Transferases ADP ribosylation Address Adenosine Diphosphate Ribose Affect Antiviral Agents Antiviral Response Antiviral Therapy Automobile Driving Biological Process Biology Cell Culture Techniques Cells Chiroptera Communicable Diseases Coronavirus Family Foundations Goals Hydrolase Hypersensitivity Immune response Impairment Infection Innate Immune Response Interferons Knock-out Knowledge Lead Malignant Neoplasms Mediating Mission Modeling Murine hepatitis virus National Institute of General Medical Sciences Pathway interactions Phenotype Phosphorylation Post-Translational Protein Processing Process Proteins RNA replication Research Research Personnel Role System Togaviridae Translations Viral Viral hepatitis Virus Virus Diseases Virus Replication Work human disease immunoregulation inhibitor/antagonist innate immune mechanisms innovation mutant novel pathogenic virus protein function response

项目摘要

PROJECT SUMMARY ADP-ribosylation is an important post-translational modification that directly influences several biological processes including cancer, allergy, and infectious disease. ADP-ribose can be added to proteins as one or more consecutive units by ADP-ribosyltransferases, also termed PARPs, resulting in mono- or poly- ADP-ribosylation (mAR or pAR). Most mARylating PARPs are upregulated by interferon (IFN) upon virus infection and several are predicted to have antiviral functions. In addition, several viral families, including the Coronaviridae and Togaviridae, encode for macrodomain proteins that have mono-ADP-ribosyl hydrolase (ARH) activity. This activity allows these viruses the ability to specifically counteract the effects of mAR, further implicating mAR in the mammalian antiviral response. Despite these findings, there are only a few known examples where mAR is known to inhibit virus replication. This is largely due to the lack of cell culture models of virus infections where the mAR status of a cell can be specifically controlled, such as models using mutant viruses or PARP knockout cells that have significant phenotypes. Importantly, the PI has established a virus infection system using a model coronavirus, Murine Hepatitis Virus (MHV), where virus lacking ARH activity is i) significantly impaired in virus replication and ii) independently induces a robust IFN response. These phenotypes are reversed by PARP inhibitors, establishing mAR as a key factor driving this anti-viral response. The investigator’s long-term goal is to determine mechanistically how mAR inhibits virus replication and enhances the innate immune response following virus infection. This gap in knowledge will be resolved by answering the following questions: 1) How does mAR inhibit MHV infection? Does it inhibit the entry, RNA replication, protein translation, assembly, or release of MHV? 2) What step(s) of the IFN induction pathway is enhanced by mAR, and does mAR also affect the IFN response in bats, which are known to harbor many highly pathogenic viruses? 3) What proteins are modified by PARPs following virus infection and which substrates are relevant for specific phenotypes? The rationale for this research is that it will enhance our understanding of mAR, including its ability to modulate protein function and will uncover novel cellular proteins or processes that mediate virus replication. The work is innovative because: i) it will bridge a significant knowledge gap between ADP-ribose biology, the innate immune response, and virus replication; ii) it utilizes unique models of infection utilizing both mutant viruses and PARP knockout cells; and iii) will be the first to address the role of mAR in bats. Finally, these projects are significant and relevant to the NIGMS mission because they will provide a thorough understanding of how mAR impacts the anti-viral response that could lay the foundation for advances in the treatment of virus infections or other human diseases impacted by mAR.

项目概要 ADP-核糖基化是一种重要的翻译后修饰，直接影响多个包括癌症、过敏和传染病在内的生物过程可以添加到蛋白质中。 ADP-核糖基转移酶（也称为 PARP）通过一个或多个连续单元，产生单或多聚 ADP-核糖基化（mAR 或 pAR）在病毒作用下会被干扰素 (IFN) 上调。此外，一些病毒家族，包括病毒，预计具有抗病毒功能。冠状病毒科和披膜病毒科，编码具有单 ADP-核糖基水解酶的大结构域蛋白 (ARH) 活性使这些病毒能够特异性抵抗 mAR 的影响，进一步。尽管有这些发现，但已知的只有少数。已知 mAR 抑制病毒复制的例子这主要是由于缺乏细胞培养模型。可以专门控制细胞的 mAR 状态的病毒感染，例如使用突变体的模型重要的是，PI 已经建立了一种病毒。使用模型冠状病毒、鼠肝炎病毒 (MHV) 的感染系统，其中缺乏 ARH 活性的病毒 i) 病毒复制显着受损，ii) 独立诱导强烈的 IFN 反应。 PARP 抑制剂可逆转表型，从而使 mAR 成为驱动这种抗病毒反应的关键因素。研究人员的长期目标是从机制上确定 mAR 如何抑制病毒复制和增强病毒感染后的先天免疫反应将通过以下方式解决这一知识空白。回答以下问题：1）mAR如何抑制MHV感染？它是否抑制RNA的进入？ MHV 的复制、蛋白质翻译、组装或释放？ 2) IFN 诱导途径的步骤是什么？ mAR 增强，并且 mAR 是否也会影响蝙蝠的 IFN 反应，众所周知，蝙蝠体内含有许多干扰素。 3) 病毒感染后哪些蛋白质会被 PARP 修饰？底物与特定表型相关吗？这项研究的基本原理是它将增强我们的研究能力。了解 mAR，包括其调节蛋白质功能的能力，并将发现新的细胞蛋白质这项工作具有创新性，因为：i）它将在一个重要的过程中发挥桥梁作用。 ADP-核糖生物学、先天免疫反应和病毒复制之间的知识差距；利用突变病毒和 PARP 敲除细胞的独特感染模型；以及 iii) 将是第一个最后，这些项目非常重要并且与 NIGMS 任务相关。因为他们将全面了解 mAR 如何影响抗病毒反应，从而可能奠定为治疗受 mAR 影响的病毒感染或其他人类疾病的进展奠定基础。