STING-dependent activation of Natural Killer cells by viral and tumor DNA

病毒和肿瘤 DNA 对自然杀伤细胞的 STING 依赖性激活

• 批准号: 9895895
• 负责人: DAVID H RAULET
• 金额: $ 33.41万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2020-03-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9895895
• 关键词: Address; Alleles; Antigen-Presenting Cells; Antitumor Response; Antiviral Agents; Antiviral Response; Binding; Bone Marrow; CRISPR interference; Cancer Model; Cells; Chimera organism; Critical Pathways; DNA; DNA Viruses; DNA biosynthesis; Data; Defect; Dendritic Cells; Dinucleoside Phosphates; Enzymes; Event; Failure; Genetic Engineering; Genetic Transcription; Genetically Engineered Mouse; Graft Rejection; Herpesviridae; Immune; Immune response; Immune signaling; In Vitro; Infection; Injections; Interferon Type I; Interferons; Investigation; Knockout Mice; Malignant Neoplasms; Mammalian Cell; Mediating; Mediator of activation protein; Membrane Transport Proteins; Minor; Modeling; Mus; NK Cell Activation; Natural Killer Cells; Neoplasm Transplantation; Pathway interactions; Periodicity; Play; Production; Proteins; Research; Role; SLC19A1 gene; Series; Shapes; Signal Pathway; Signal Transduction; Site; Sting Injury; T cell response; T-Lymphocyte; Testing; Viral Cancer; Virus; Virus Diseases; anti-tumor immune response; base; cancer cell; cancer immunotherapy; cell transformation; cytokine; genetic approach; genome-wide; immune activation; in vivo; inhibitor/antagonist; intercellular communication; macrophage; neoplastic cell; neutrophil; novel strategies; pathogen; prevent; receptor; response; sarcoma; small molecule; treatment response; tumor; tumor DNA; tumor microenvironment; viral DNA

The cGAS-STING pathway senses cytosolic DNA in infected cells and cancer cells and triggers production of type I interferon and other cytokines, thus triggering immune responses. Deficiency in the pathway prevents T cell responses to several viruses and to transplanted tumor lines. Our new results show that the pathway is critical for activating spontaneous NK cell responses against cancer and virus infections. Furthermore, in probing the underlying mechanisms, our research has uncovered a new cellular mechanism whereby cGAS and STING activate immune responses. Using genetic approaches, we found that the NK response to transferred tumor cells depends on cGAS expression in tumor cells and STING expression in host cells. We interpret these data to indicate that cGAS activation is the key event that occurs in infected or transformed cells resulting in the production of the cyclic dinucleotide cGAMP. cGAMP is then transferred from the infected/transformed cells to other immune cells, such as antigen presenting cells, which are induced to initiate the immune response. Hence, we propose that cGAMP transfer between cells is a fundamental mechanism in immune activation. In considering how cGAMP is transferred between cells, we hypothesized that specific membrane transporters must play a role. Using a genome-wide CRISPRi screen, we identified two transporter molecules that specifically import cGAMP into cells, one of which plays a major role and other a minor role. Using a series of knockout mice and conditional knockout mice, and cellular manipulations and transfers, we propose to test the requirement of cGAS-STING signaling in NK and T cell responses to cancer and DNA viruses, the generality of the requirement of the transfer mechanism in viral infections and cancer models and for T cell and NK cell responses, including the roles of the newly identified transporters, and to define the specific cells that must import the cGAMP for immune responses to occur. Specifically, we will: (1) Determine the sites of action of cGAMP and STING and intercellular transfer of cGAMP in anti-viral responses, including NK and T cell responses; (2) Determine sites of action of cGAMP and STING and intercellular transfer of cGAMP in T cell responses to cancer, in transfer models and GEM cancer models; (3) Determine roles of cGAMP transporters in anti-viral and anti-tumor and immune responses. Using conditional knockout mice and inhibitor studies, we will test their function in anti-herpesvirus and anti-tumor responses, including a GEM model.

CGAS-STING通路感应感染细胞和癌细胞中的胞浆DNA，并触发 I型干扰素和其他细胞因子，从而触发免疫反应。该途径的缺陷会阻止T 细胞对几种病毒和移植的肿瘤细胞的反应。我们的新结果表明，这条途径是 对于激活自然杀伤细胞对抗癌症和病毒感染的自发反应至关重要。此外，在 通过对潜在机制的探索，我们的研究发现了一种新的细胞机制，即cGAS 刺痛会激活免疫反应。使用遗传方法，我们发现NK对 转移的肿瘤细胞依赖于肿瘤细胞中cGAS的表达和宿主细胞中的STIN表达。我们 解释这些数据以表明cGAS激活是发生在感染或转化细胞中的关键事件 从而产生环二核苷酸cGAMP。然后将cGAMP从 被感染/转化的细胞转化为其他免疫细胞，如抗原提呈细胞，被诱导启动 免疫反应。因此，我们认为cGAMP在细胞间的转移是一种基本的 免疫激活机制。在考虑cGAMP如何在细胞间转移时，我们假设 认为特定的膜转运蛋白必须发挥作用。使用全基因组CRISPRi筛查，我们确定了两个 特异性地将cGAMP导入细胞的转运体分子，其中一个起主要作用，另一个起主要作用 次要角色。使用一系列基因敲除小鼠和条件基因敲除小鼠，以及细胞操作和 转移，我们建议测试cGAS-STING信号在NK和T细胞对癌症的反应中的需求 和DNA病毒一样，病毒感染和癌症转移机制的共性要求 模型和T细胞和NK细胞反应，包括新发现的转运蛋白的作用，以及 定义必须导入cGAMP才能产生免疫反应的特定细胞。具体来说，我们会： (1)确定cGAMP的作用部位和cGAMP的刺痛和细胞间转移 反应，包括NK和T细胞反应；(2)确定cGAMP和STIN的作用部位和 在转移模型和GEM癌模型中，cGAMP在T细胞对癌症的反应中的细胞间转移； (3)确定cGAMP转运蛋白在抗病毒、抗肿瘤和免疫应答中的作用。vbl.使用 条件性基因敲除小鼠和抑制剂研究，我们将测试它们在抗疱疹病毒和抗肿瘤方面的功能 回应，包括创业板模型。