Mechanistic Exploration of cGAS-STING-Mediated Vaccine Enhancement

cGAS-STING 介导的疫苗增强机制探索

基本信息

批准号：
10318966
负责人：
VICTOR Robert DEFILIPPIS
金额：
$ 78.71万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-01-23 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10318966
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 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 Immunization Immunologics Immunotherapeutic agent In Vitro Infection Innate Immune Response Ligands Link Lymphoid Cell Macaca mulatta Mediating Messenger RNA Microbe Mitochondria Modeling Molecular Mus Myeloid Cells Nuclear Ontology Outcome Pathway interactions Pattern Pattern recognition receptor Penetration Periodicity Pharmacology Phenotype Physiological Processes Primates Process Proteins Research Role Signal Transduction Signaling Protein Solubility Stimulator of Interferon Genes Stromal Cells 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 recruit response small molecule tool transcription factor transcriptomics vaccination outcome vaccine efficacy

项目摘要

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.

项目摘要该R01提案的目标包括疫苗调整的分子和免疫学表征与干扰素基因蛋白刺激剂介导的过程相关的活性（STING）。两个重要目前，不足是有关辅助人类疫苗的证据。首先，很少有调节器第二个在美国批准用于临床用途，第二个是调整相关免疫的确切机械基础增强知之甚少。有效的调节器触发快速，局部的先天免疫反应在他们的管理之后。从根本上讲，先天信号是通过模式的参与来启动的指示或模仿微生物感染的识别受体（PRR）。反过来，这导致了免疫调节和促炎性因子的表达最终导致适应性免疫回报能够消除感染组织。 sting代表感受循环二核苷酸（CDN）的PRR 细胞酶GMP-AMP合酶（CGA）的乘积在检测到细胞质dsDNA之后源自微生物，线粒体或细胞核。刺激介导的表型最终由由活化蛋白转录诱导的基因。这完全涉及转录因子 IFN调节因子3（IRF3）和核因子κB（NF-κB），它们合成不同的本体学的mRNA 其功能角色大多是出乎意料的。此外，Sting似乎控制了物理过程免疫系统的细胞类型之间的不同方式，包括基质，髓样和T细胞。有趣的是，在鼠模型中，药理学诱导sting依赖性活性大大提高了疫苗效率由对潜水病原体引起的受保护的免疫力指示。不幸的是，精确的分子和与刺激活性相关的适应性免疫酮增强的先天相关性在很大程度上尚未进行。此外，刺痛的调节器是否引起私人的有效免疫原性结果检查。我们计划将这些与强大的CRISPR和转录组技术相结合，以获得对刺激介导的免疫结局的基本基础的渗透性洞察力。我们假设在功能上，增强与基于STING的调节器相关的抗原指导的适应性免疫学是功能与使用这些模型可分辨的分子和细胞过程有关。我们还确定了一流的小分子，可以激活跨物种的CGAS，并增强对Zika的免疫原性病毒抗原。在体外，鼠，鼠和非人类灵长类动物（NHP）模型中包括在内，将允许美国至：1）验证和将CGA描述为新的免疫治疗靶标； 2）演示刺痛调整高度临床相关的模型物种的效率； 3）确定规格特定的相似性和差异关于刺激介导的免疫反应。