ZFAND6 regulation of innate antiviral immunity

ZFAND6 对先天抗病毒免疫的调节

基本信息

批准号：
9979076
负责人：
EDWARD W HARHAJ
金额：
$ 23.75万
依托单位：
PENNSYLVANIA STATE UNIV HERSHEY MED CTR
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：
2020-03-16 至 2022-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9979076
关键词：
A20 protein Address Agonist Antiviral Agents Automobile Driving Binding Biological Bone Marrow C-terminal Cachexia Cells DNA DNA Viruses Development Dose Down-Regulation Enzymes Family Family member Feedback Fibroblasts Foundations Frequencies Gene Targeting Genes Herpesvirus 1 Homeostasis Human Genome IRF1 gene IRF3 gene Immune Immune signaling Infection Inflammation Inflammatory Inflammatory Response Influenza A virus Innate Immune Response Interferon Type I Interferons Investigation Knockout Mice Knowledge Link Lung Lung Inflammation Lysine Mediating Molecular Morbidity - disease rate Mus N-terminal Natural Immunity Nucleic Acids PRKCA gene Pathogenicity Pathway interactions Pattern Pattern recognition receptor Phosphotransferases Physiological Plants Play Polyubiquitin Proteins Publishing Pulmonary Inflammation RNA RNA Viruses Regulation Reporter Resistance Role Sequence Homology Signal Pathway Signal Transduction Signaling Protein Spleen Stains Stimulus Stress-Induced Protein Tissues Toll-like receptors Transferase Ubiquitin Vesicular stomatitis Indiana virus Viral Viral Load result Virus Virus Diseases Wild Type Mouse Zinc Fingers antiviral immunity cell type chemokine cytokine experimental study fitness gene induction gut microbiota helicase immune activation immunopathology insight macrophage mortality novel therapeutics ovarian neoplasm overexpression pathogen pathogenic virus prevent protein degradation protein function reconstitution repaired response stress tolerance transcription factor transcriptome sequencing viral DNA

项目摘要

ABSTRACT The A20 ubiquitin-editing enzyme is a key negative regulator of NF-κB and innate immune signaling pathways. A20 contains seven zinc finger (ZF1-7) domains, of which ZF4 interacts with lysine 63 (K63)-linked polyubiquitin chains to suppress NF-κB signaling. Including A20, there are 10 genes containing A20 ZF domains encoded in the human genome; however, with the exception of A20 the functions of these proteins are poorly understood. ZFAND6 (also known as AWP1) contains an A20-like ZF domain that shares significant sequence homology with A20 ZF4, and also harbors an AN1-type ZF. Although published overexpression studies suggest that ZFAND6 may inhibit NF-κB, the specific pathways regulated by ZFAND6 and its physiological roles have remained elusive. In preliminary studies for this exploratory proposal, we have used gene targeting to generate mice lacking the Zfand6 gene. Zfand6–/– mice are born at expected Mendelian frequencies with no obvious developmental or immune abnormalities. RNA sequencing of Zfand6–/– bone marrow-derived macrophages revealed spontaneous induction of interferon-stimulated genes (ISGs), which rendered these cells highly resistant to infection with a broad range of DNA and RNA viruses. The DNA sensing pathway adaptor STING and the antiviral transcription factor IRF1 were both upregulated in Zfand6–/– cells, which may drive the resistance of ZFAND6-deficient cells to virus infection. ISGs were upregulated in the spleen, but not in the lungs, of Zfand6–/– mice, and knockout mice were more susceptible than wild-type mice to mortality upon challenge with a low dose of influenza A virus (IAV) despite comparable viral loads. Therefore, loss of ZFAND6 may perturb immune homeostasis and exacerbate virus-induced inflammation and immunopathology. The central hypothesis driving these investigations is that ZFAND6 targets key innate immune signaling proteins for degradation to prevent the spontaneous induction of ISGs which may contribute to overexuberant inflammation triggered by virus infection. To address this hypothesis we will perform the following Specific Aims: 1) Determine the mechanisms of ZFAND6 inhibition of antiviral signaling, and 2) Determine the role of ZFAND6 in regulating the inflammatory response to IAV infection. Completion of these studies will define a physiological role for ZFAND6 in restricting innate antiviral immunity and expand our knowledge on the mechanisms of immune homeostasis which serve to mitigate virus-induced inflammation.

抽象的 A20泛素编辑酶是NF-κB和先天免疫信号通路的关键负调节剂。 A20包含七个锌指（ZF1-7）域，其中ZF4与赖氨酸63（K63）链接的相互作用多泛素链以抑制NF-κB信号传导。包括A20，有10个含有A20 ZF的基因人类基因组中编码的领域；但是，除A20以外，这些蛋白质的功能知之甚少。 ZFAND6（也称为AWP1）包含一个类似A20的ZF域，具有重要的与A20 ZF4的序列同源性，还拥有AN1型ZF。尽管出版了过表达研究表明，ZFAND6可能抑制NF-κB，NF-κB是由Zfand6及其调节的特定途径生理角色仍然难以捉摸。在此探索性建议的初步研究中，我们使用了基因靶向产生缺乏ZFAND6基因的小鼠。 ZFAND6 - / - 老鼠出生于预期的门德利安 ZFAND6 - / - 骨的RNA测序骨髓来源的巨噬细胞显示干扰素刺激的基因（ISGS）的赞助，这些诱导症使这些细胞高度抗性地具有多种DNA和RNA病毒感染。 DNA ZFAND6 - / - 都更新了传感途径适配器的刺激和抗病毒转录因子IRF1 细胞，可能会驱动ZFAND6缺陷细胞对病毒感染的抗性。 ISG已在 zfand6 - / - 小鼠和淘汰小鼠的脾脏，但不在肺中，比野生型小鼠更容易受到影响挑战时死亡率低，剂量低剂量的病毒（IAV）所需的可比病毒载量。因此，ZFAND6的丧失可能会扰动免疫稳态并加剧病毒诱导的感染和免疫病理学。推动这些调查的中心假设是ZFAND6针对关键先天免疫信号蛋白用于降解，以防止ISG的赞助诱导可能有助于通过病毒感染触发的过度检查感染。为了解决这一假设，我们将执行以下特定目的：1）确定ZFAND6抑制抗病毒信号的机制，以及2）确定ZFAND6在确定对IAV感染的炎症反应中的作用。这些完成研究将定义ZFAND6在限制先天抗病毒免疫学上的身体作用并扩展我们有关免疫稳态机制的知识，可减轻病毒诱发的炎症。