Unlocking evolutionarily latent immune functions for treating disease

项目总结/摘要 所有动物都具有强大的先天免疫反应，这取决于它们识别病毒双链的能力。 单链RNA（dsRNA）作为外源。然而，动物细胞也编码和表达dsRNA，这种细胞dsRNA 必须区分为"自身"以防止异常免疫反应。腺苷脱氨酶， RNA或ADAR使dsRNA脱氨基以将其标记为自身并防止异常免疫应答。以此身份 ADAR充当"先天免疫检查点"（IIC），并且最近的研究表明，阿达尔活性的降低 在肿瘤中释放这种IIC，引发导致细胞死亡的免疫反应。ADAR是唯一已知的IIC 迄今为止，拟议的研究旨在填补这一知识空白，以实现新的目标。 免疫疗法比较系统发育分析将与分子生物学和 生物化学实验，以确定缓解措施，如ADAR，防止不适当的部署 抗病毒防御和古老的不相容性，如无脊椎动物蛋白质，可以激活抗病毒药物， 当被引入脊椎动物时的反应。在哺乳动物细胞、小鼠和无脊椎动物中的实验 模式生物C.线虫，将提供一个广泛的系统发育抽样，以确定，测试和比较新的IIC。 工程小鼠和细胞系已经在手，并且建立了检测方法，以监测对免疫系统的影响。 两种动物的路径。已知的dsRNA结合蛋白，以及通过免疫沉淀鉴定的那些 战略，将优先进行系统发育分析测试作为IIC。体外生物化学实验，以及 结构分析，将指导随后几轮的系统发育比较。哺乳动物ADAR 1 p150 通过调节脊椎动物先天免疫途径的MDA 5臂来防止干扰素应答，和 尚未报告RIG-I组的IIC。识别RIG-I臂IIC的策略将集中在酶上 已知修饰RNA的5 '末端，其为RIG-I识别的已知表位。建议的高潮 研究将是在肿瘤发生的实验模型中评价候选IIC。

Project Summary/Abstract All animals possess a robust innate immune response that depends on their ability to recognize viral double- stranded RNA (dsRNA) as foreign. Yet, animal cells also encode and express dsRNA, and this cellular dsRNA must be distinguished as “self” to prevent an aberrant immune response. Adenosine deaminases that act on RNA, or ADARs, deaminate dsRNA to mark it as self and prevent an aberrant immune response. In this capacity ADARs serve as an ”Innate Immune Checkpoint” (IIC), and recent studies reveal that a decrease in ADAR activity in tumors releases this IIC, eliciting an immune response that leads to cell death. ADARs are the only IIC known to date, and proposed studies are designed to fill this gap in knowledge towards the goal of new immunotherapies. Comparative phylogenetic analyses will be complemented with molecular biology and biochemistry experiments to identify mitigators, such as ADARs, that prevent inappropriate deployment of antiviral defense, and ancient incompatibilities, such as invertebrate proteins that may activate an antiviral response when introduced into vertebrates. Experiments in mammalian cells and mice, and the invertebrate model organism, C. elegans, will provide a wide phylogenetic sampling to identify, test, and compare new IICs. Engineered mice and cell lines are in hand, and established assays are in place, to monitor effects on the immune pathway of both animals. Known dsRNA binding proteins, as well as those identified by immunoprecipitation strategies, will be prioritized by phylogenetic assays for testing as IICs. In vitro biochemistry experiments, and structural analyses, will guide subsequent rounds of phylogenetic comparisons. Mammalian ADAR1 p150 prevents an interferon response by modulating the MDA5 arm of the vertebrate innate immune pathway, and IICs for the RIG-I arm have not been reported. Strategies to identify IICs for the RIG-I arm will focus on enzymes known to modify the 5' terminus of RNA, a known epitope for RIG-I recognition. The culmination of proposed studies will be the evaluation of candidate IICs in experimental models of tumorigenesis.