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：
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
项目类别：
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

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 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，从而触发免疫反应。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自发性NK细胞反应针对癌症和病毒感染至关重要。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，我们确定了两个专门将CGAMP进口到细胞的转运蛋白分子，其中一种起主要作用，另一种作用次要角色。使用一系列敲除小鼠和有条件的敲除小鼠，以及细胞操纵和转移，我们建议测试NK和T细胞对癌症的CGAS丁字传导的需求和DNA病毒，病毒感染和癌症中转移机制需求的一般性模型以及T细胞和NK细胞响应，包括新鉴定的转运蛋白的作用，以及定义必须导入CGAMP以进行免疫反应发生的特定细胞。具体来说，我们将：（1）确定抗病毒中CGAMP，刺激和细胞间转移的作用部位响应，包括NK和T细胞反应；（2）确定CGAMP和STING的作用部位以及在转移模型和GEM癌症模型中，CGAMP在T细胞反应中CGAMP的细胞间转移；（3）确定CGAMP转运蛋白在抗病毒和抗肿瘤和免疫反应中的作用。使用有条件的基因敲除小鼠和抑制剂研究，我们将测试它们在抗herpesvirus和抗肿瘤中的功能响应，包括宝石模型。