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

基本信息

批准号：
10561647
负责人：
DAVID H RAULET
金额：
$ 45.3万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-06-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10561647
关键词：
Alleles Antigen-Presenting Cells Antitumor Response Antiviral Response Binding Bone Marrow CRISPR interference Cancer Model Cells Chimera organism Cre driver Critical Pathways DNA DNA Viruses DNA biosynthesis Data Data Analyses 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 Macrophage Malignant Neoplasms Mammalian Cell Mediating Mediator Membrane Transport Proteins Modeling Mus NK Cell Activation Natural Killer Cells Neoplasm Transplantation Pathway interactions Periodicity Process Production Productivity Proteins Research Role SLC19A1 gene Series Shapes Signal Pathway Signal Transduction Site Stimulator of Interferon Genes Sting Injury T cell response T-Lymphocyte Testing Viral Viral Cancer Virus Virus Diseases anti-tumor immune response cancer cell cancer immunotherapy cell transformation conditional knockout cytokine genetic approach genome-wide immune activation in vivo inhibitor intercellular communication neoplastic cell neutrophil novel novel strategies pathogen prevent receptor response small molecule treatment response tumor tumor DNA tumor microenvironment viral DNA

项目摘要

The cGAS-STING pathway senses cytosolic DNA in infected cells and cancer cells and triggers production of type I interferon and other cytokines, thus eliciting 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 a transporter molecule that specifically imports cGAMP into cells. 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, the roles of the newly identified transporters in the process, 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 of cancer.

CGAS丁字道在感染细胞和癌细胞中感觉到胞质DNA，并触发产生 I型干扰素和其他细胞因子，从而引起免疫反应。路径的不足阻止细胞对几种病毒和移植肿瘤线的反应。我们的新结果表明该路径是激活针对癌症和病毒感染的自发性NK细胞反应至关重要。此外，在探测潜在机制，我们的研究发现了一种新的细胞机制刺痛激活免疫反应。使用遗传方法，我们发现NK对转移的肿瘤细胞取决于肿瘤细胞中的CGA表达和宿主细胞中的刺激表达。我们解释这些数据以表明CGA激活是在感染或转化的单元格中发生的关键事件导致循环二核苷酸CGAMP的产生。然后将CGAMP从被感染/转化为其他免疫细胞的细胞，例如抗原呈递细胞，这些细胞被诱导启动免疫反应。因此，我们建议CGAMP在细胞之间转移是一个基本免疫激活中的机制。在考虑CGAMP如何在细胞之间转移时，我们假设该特定的膜转运蛋白必须发挥作用。使用全基因组CRISPRI屏幕，我们确定了专门将CGAMP进口到细胞的转运蛋白分子。使用一系列淘汰小鼠，有条件的敲除小鼠以及细胞操作和转移，我们建议测试 NK和T细胞对癌症和DNA病毒的反应中的CGAS刺信信号传导，在病毒感染和癌症模型以及T细胞和NK细胞中转移机制的要求响应，新鉴定的转运蛋白在此过程中的作用，并定义特定细胞必须导入CGAMP以进行免疫反应。具体来说，我们将：（1）确定抗病毒反应中CGAMP的CGAMP，刺激和细胞间转移的作用，包括NK 和T细胞反应；（2）确定CGAMP，STING和细胞间转移的作用部位 T细胞对癌症的CGAMP，转移模型和GEM癌模型中的CGAMP；（3）确定角色抗病毒和抗肿瘤和免疫反应中的CGAMP转运蛋白。使用有条件的淘汰小鼠和抑制剂研究，我们将测试它们在抗herpesvirus和抗肿瘤反应中的功能，包括GEM 癌症模型。