Harnessing ZBP1-driven cell death to improve influenza vaccine efficacy

利用 ZBP1 驱动的细胞死亡来提高流感疫苗的功效

• 批准号: 10455196
• 负责人: SIDDHARTH BALACHANDRAN
• 金额: $ 84.79万
• 依托单位: RESEARCH INST OF FOX CHASE CAN CTR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-13 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10455196
• 关键词: Adjuvant; Adopted; Agonist; Antigens; Apoptosis; Attenuated Vaccines; CD8-Positive T-Lymphocytes; Cell Death; Cell Death Signaling Process; Cell Nucleus; Cells; Cellular Immunity; Cessation of life; Cues; Cytoplasm; Cytotoxic T-Lymphocytes; Defective Viruses; Double-Stranded RNA; Engineering; Epithelial Cells; Extracellular Space; Goals; Hand; Hospitalization; Host Defense; Humoral Immunities; Immune; Immune response; Immunity; Immunologics; Individual; Inflammatory; Influenza A Virus, H1N1 Subtype; Influenza A Virus, H3N2 Subtype; Influenza A virus; Left; Ligands; Lung; Lytic Virus; Mediating; Modality; Modeling; Molecular Conformation; Necrosis; Nuclear; Nuclear Envelope; Nucleoproteins; Pathogenesis; Pathologic; Pathway interactions; Positioning Attribute; Proteins; RNA; RNA Sequences; Recombinants; Reporting; Role; Rupture; Signal Pathway; Source; Stromal Cells; Structure; T cell response; T-Lymphocyte; Technology; Testing; Vaccination; Vaccines; Viral Antigens; Viral Genome; Viral Proteins; Virus; Virus Diseases; Virus Replication; adaptive immune response; adaptive immunity; airway epithelium; anti-influenza; base; cell type; cytotoxic CD8 T cells; design; experimental study; genomic RNA; immunogenic; immunogenic cell death; improved; in vivo; influenza virus vaccine; lymph nodes; macrophage; neutrophil; next generation; novel strategies; novel vaccines; prevent; response; seasonal influenza; sensor; universal influenza vaccine; vaccine efficacy; vaccine response

Project Summary Vaccination represents the most effective means of preventing influenza A virus (IAV) infections. Current approaches to IAV vaccines primarily target the HA protein, but HA undergoes significant antigenic drift and is thus a poor target for lasting humoral immunity. An alternative approach to developing a universal IAV vaccine is to target internal viral proteins (such as the nucleoprotein), which are also highly conserved between most IAV strains. A major roadblock to this approach, however, is a lack of adjuvants that trigger the right kind of immune response - a ‘type I’ (or Th1/CD8+ T-cell driven) response - to these internal antigens. In this proposal, we outline a new strategy that seeks to harness immunogenic cell death as a means of activating type I immunity to IAV vaccines. We have identified a host signaling pathway which accounts for almost all IAV-activated immunogenic death in infected cells, including primary airway epithelial cells, macrophages and DCs. This cell death pathway is initiated when the host sensor protein ZBP1 detects IAV genomic RNA and activates parallel pathways of programmed necrosis (necroptosis) and apoptosis to kill the infected cell and instigate CD8+ T cell-mediated adaptive immune responses. Adding to the attractiveness of ZBP1-initiated cell death as a vaccine approach are our discoveries, first, that IAV produces a new form of double-stranded RNA - termed Z-RNA - to activate ZBP1. Z-RNA is thus a new PAMP, and can be readily harnessed to trigger immunogenic cell death and adjuvant IAV vaccines. Second, ZBP1 initiates necroptosis from the nucleus, resulting in nuclear envelope rupture and release of highly-immunogenic nuclear DAMPs and IAV antigens. Nuclear necroptosis is even more immunogenic than conventional (cytoplasm-initiated) necroptosis. As ZBP1-driven immunogenic cell death (both apoptosis and nuclear necroptosis) is important for effective anti-IAV cytotoxic T cell responses, our discoveries allow us to propose that Z-RNAs are potent new adjuvants for activating type I immune responses to IAV, and that recombinant IAVs engineered to produce Z-RNAs represent new type I immunity-activating vaccines. In this proposal, three labs with strong expertise in IAV cell death signaling (Balachandran), recombinant IAV technology (Langlois), and pathogenesis/vaccine strategies (Lopez) will unite to identify IAV-generated Z-RNAs that activate ZBP1 (Aim 1); determine which stromal and immune cell types die by ZBP1-mediated mechanisms to initiate and regulate effective adaptive immune responses (Aim 2); and test if triggering ZBP1-mediated cell death with synthetic and virus-generated Z-RNAs can be leveraged to inform and supply next-generation vaccines capable of providing effective cross-protective immunity to IAV (Aim 3).

项目摘要 疫苗接种是预防影响病毒（IAV）感染的最有效手段。当前的 IAV疫苗主要靶向HA蛋白的方法，但HA经历了明显的抗原漂移，IS 因此，持久的体液免疫史的目标很差。开发通用IAV疫苗的另一种方法 是靶向内部病毒蛋白（例如核蛋白），它们在大多数IAV之间也高度保守 菌株。但是，这种方法的主要障碍是缺乏触发正确免疫的调节器 响应 - 对这些内部抗原的“ I型I”（或TH1/CD8+ T细胞驱动器）响应。在此提案中，我们概述了 一种试图利用免疫原性细胞死亡作为激活I型免疫（IS）的一种新策略 疫苗。我们已经确定了一种宿主信号通路，该途径几乎占所有IAV激活的免疫原性 感染细胞的死亡，包括原发性气道上皮细胞，巨噬细胞和DC。这个细胞死亡途径 当宿主传感器蛋白ZBP1检测IAV基因组RNA并激活平行途径时，开始 程序性坏死（坏死）和凋亡杀死受感染的细胞并激发CD8+ T细胞介导的 自适应免疫反应。增加ZBP1引起的细胞死亡作为疫苗方法的吸引力是 首先，我们的发现IAV产生了一种新形式的双链RNA（称为Z -RNA），以激活ZBP1。 Z-RNA因此是一种新的弹药，可以很容易地利用以触发免疫原性死亡并调整IAV 疫苗。其次，ZBP1从核启动坏死，导致核包膜破裂并释放 核坏死性甚至比高度免疫原性核潮湿和IAV抗原更具免疫原性。 常规（细胞质引起的）坏死性。作为ZBP1驱动的免疫原性细胞死亡（凋亡和 核坏死性）对于有效的抗IAV细胞毒性T细胞反应很重要，我们的发现使我们能够 提议Z-RNA是激活I型免疫反应IAV的潜在新调节器，并且 设计用于产生Z-RNA的重组IAV代表了新型I型免疫激活疫苗。在这个 提案，三个在IAV细胞死亡信号传导（Balachandran），重组IAV方面具有强大专业知识的实验室 技术（Langlois）和发病机理/疫苗策略（Lopez）将团结起来以识别IAV生成的Z-RNA 激活ZBP1（AIM 1）；确定ZBP1介导的机制死亡哪种基质和免疫电池类型 启动和调节有效的适应性免疫回报（AIM 2）；并测试是否触发ZBP1介导的细胞 可以利用合成和病毒生成的Z-RNA的死亡来告知和提供下一代 能够为IAV提供有效的跨保护免疫学的疫苗（AIM 3）。