Mechanistic Exploration of cGAS-STING-Mediated Vaccine Enhancement

cGAS-STING 介导的疫苗增强机制探索

• 批准号: 10534676
• 负责人: VICTOR Robert DEFILIPPIS
• 金额: $ 77.3万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-23 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10534676
• 关键词: Adjuvant; Agonist; Animal Model; Antigen-Presenting Cells; Antigens; Biological; Biology; CRISPR/Cas technology; Categories; Cell Line; Cell Nucleus; Cell physiology; Cells; Chemicals; Chemosensitization; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Cyclic GMP; Cytoplasm; Detection; Dinucleoside Phosphates; Dissection; Enzymes; Evaluation; Exhibits; Experimental Models; Formulation; Gene Activation; Genes; Genetic; Genetic Transcription; Goals; Human; IRF3 gene; Immune; Immune response; Immune system; Immunity; Immunologic Stimulation; Immunologics; Immunotherapeutic agent; In Vitro; Infection; Inflammatory; Innate Immune Response; Ligands; Link; Lymphoid Cell; Macaca mulatta; Mediating; Messenger RNA; Microbe; Mitochondria; Modeling; Molecular; Mus; Myelogenous; Myeloid Cells; Nuclear; Ontology; Outcome; Pathway interactions; Pattern; Pattern recognition receptor; Periodicity; Phenotype; Physiological Processes; Primates; Process; Proteins; Research; Role; Signal Transduction; Signaling Protein; Solubility; Stimulator of Interferon Genes; System; T-Lymphocyte; Technology; Testing; Tissues; Transcript; Translating; Vaccination; Vaccine Adjuvant; Vaccines; Viral Antigens; Zika Virus; adaptive immune response; adaptive immunity; analog; base; cell type; clinically relevant; ds-DNA; gene function; human model; immunogenic; immunogenicity; immunoregulation; in vivo; innate immune function; insight; microbial; monocyte; mouse model; nonhuman primate; novel; pathogen; pathogenic microbe; pharmacologic; recruit; response; small molecule; tool; transcription factor; transcriptomics; vaccination outcome; vaccine efficacy

PROJECT SUMMARY The goals of this R01 proposal include the molecular and immunological characterization of vaccine adjuvant activity associated with processes mediated by the protein Stimulator of Interferon Genes (STING). Two vital insufficiencies are currently evident regarding adjuvanted human vaccines. First, very few adjuvants are approved for clinical use in the U.S. Second, the precise mechanistic bases of adjuvant-associated immune augmentation are poorly understood. Effective adjuvants trigger rapid, localized innate immune responses following their administration. Fundamentally, the innate signaling is initiated through engagement of pattern recognition receptors (PRRs) by ligands indicative or imitative of microbial infection. This, in turn, leads to expression of immunomodulatory and proinflammatory factors that ultimately direct adaptive immune responses capable of eliminating infected tissues. STING represents the PRR that senses cyclic dinucleotides (CDN), a product of the cellular enzyme cyclic GMP-AMP synthase (cGAS) following its detection of cytoplasmic dsDNA derived from microbes, mitochondria, or the nucleus. STING-mediated phenotypes are ultimately conferred by genes that are transcriptionally induced by the activated protein. This crucially involves the transcription factors IFN regulatory factor 3 (IRF3) and nuclear factor κB (NF-κB), which synthesize mRNAs of distinct ontologies yet whose functional roles are mostly unexplored. Moreover, STING appears to control physiological processes that differ dramatically between cell types of the immune system including stromal, myeloid, and T cells. Intriguingly, pharmacologic induction of STING-dependent activity in murine models greatly enhances vaccine efficacy as indicated by protective immunity elicited against diverse pathogens. Unfortunately, the precise molecular and innate correlates of adaptive immune potentiation associated with STING activity remain largely unexamined. Furthermore, whether STING adjuvants elicit similarly effective immunogenic outcomes in primates has not been examined. We plan to couple these with our powerful CRISPR and transcriptomic technologies to obtain penetrative insight into the fundamental bases of STING-mediated immune outcomes. We hypothesize that the enhancement of antigen-directed adaptive immunity associated with STING-based adjuvants is functionally linked to molecular and cellular processes that are discernable using these models. We have also identified a first-in-class small molecule that activates cGAS-STING across species and enhances immunogenicity to Zika virus antigen. Including this alongside CDN in in vitro, murine, and nonhuman primate (NHP) models will allow us to: 1) Validate and characterize cGAS as a new immunotherapeutic target; 2) Demonstrate STING adjuvant efficacy in a highly clinically relevant model species; and 3) Identify species-specific similarities and differences with respect to STING-mediated immune responses.

项目概要 该 R01 提案的目标包括疫苗佐剂的分子和免疫学表征 与干扰素基因刺激蛋白 (STING) 介导的过程相关的活性。两个至关重要的 目前人类佐剂疫苗明显存在不足。首先，助剂很少 在美国获批临床使用 二、佐剂相关免疫的精确机制基础 人们对增强了解甚少。有效的佐剂触发快速、局部的先天免疫反应 在他们的管理之后。从根本上说，先天信号是通过模式的参与而启动的。 指示或模拟微生物感染的配体识别受体（PRR）。这反过来又导致 最终指导适应性免疫反应的免疫调节因子和促炎因子的表达 能够消除受感染的组织。 STING 代表感知环状二核苷酸 (CDN) 的 PRR， 细胞酶环 GMP-AMP 合酶 (cGAS) 检测细胞质 dsDNA 后的产物 源自微生物、线粒体或细胞核。 STING 介导的表型最终由 由激活的蛋白质转录诱导的基因。这关键涉及转录因子 IFN 调节因子 3 (IRF3) 和核因子 κB (NF-κB)，它们合成不同本体的 mRNA 其功能作用大多未被探索。此外，STING 似乎可以控制生理过程 免疫系统的细胞类型（包括基质细胞、骨髓细胞和 T 细胞）之间存在显着差异。有趣的是， 在小鼠模型中药理学诱导 STING 依赖性活性大大增强了疫苗功效， 通过针对不同病原体引起的保护性免疫来表明。不幸的是，精确的分子和 与 STING 活性相关的适应性免疫增强的先天相关性在很大程度上仍未得到检验。 此外，STING 佐剂是否在灵长类动物中引发类似有效的免疫原性结果尚未得到证实。 检查了。我们计划将这些与我们强大的 CRISPR 和转录组技术结合起来，以获得 深入了解 STING 介导的免疫结果的基本基础。我们假设 与基于 STING 的佐剂相关的抗原定向适应性免疫的增强在功能上是 与使用这些模型可辨别的分子和细胞过程相关联。我们还确定了一个 一流的小分子，可跨物种激活 cGAS-STING 并增强对寨卡病毒的免疫原性 病毒抗原。将其与 CDN 一起纳入体外、小鼠和非人类灵长类动物 (NHP) 模型中，将允许 我们： 1) 验证和表征 cGAS 作为新的免疫治疗靶点； 2) 演示STING佐剂 在临床高度相关的模型物种中的功效； 3) 识别特定物种的相似点和差异 关于 STING 介导的免疫反应。