Novel Role for Host Immunostimulatory RNA in Antiviral Immune Defense

项目摘要 来自许多实验室的工作已经确定，DNA传感器-膜定位 Toll样受体（例如TLR 9）和细胞内DNA传感器（例如cGAS）-正义疱疹病毒 感染与此形成鲜明对比的是，我们对细胞质内的生理相关性的认识， RIG-I样受体（RLR）家族的RNA传感器在疱疹病毒检测中的应用， 单纯疱疹病毒1型（HSV-1），是基本的。 这项拟议中的研究建立在Gack实验室最近发现的基础上，该发现表明RIG-I发挥了重要作用。 在检测和限制HSV-1感染中起关键作用，其中RIG-I和细胞内DNA 传感器以时间方式起作用。此外，我们发现，在HSV-1感染期间， 宿主来源的5S核糖体RNA假基因转录物141（RNA 5SP 141）激活RIG-I， 依来替尼介导的抗病毒先天免疫防御。在合作工作中，我们最近 发现RNA 5SP 141基因表达缺失与重度HSV-1相关， 人类的脑炎此外，我们发现RNA 5SP 141从感染的细胞中输出， 细胞通过外泌体并被未感染的细胞吸收，在那里它触发了RIG-I信号传导。 使用一系列分子、生物化学和细胞生物学方法相结合的协调方法， 通过CRISPR基因编辑技术，我们将确定RNA 5SP 141基因表达的作用， 在对HSV-1感染的抗病毒先天免疫应答和HSV-1相关疾病中 (AIM 1）。此外，我们将阐明RNA 5SP 141的分子机制 外泌体负载（AIM 2），并确定外泌体递送的生理相关性， RNA 5SP 141在宿主对HSV-1感染的免疫中的作用（AIM 3）。我们的研究将提供一个分子 了解先天免疫检测疱疹病毒感染，这可能会提供 为新的治疗方法奠定基础或指导新型佐剂的设计。

PROJECT SUMMARY Work from many laboratories has established that DNA sensors – both membrane-localized Toll-like receptors (e.g. TLR9) and intracellular DNA sensors (e.g. cGAS) – sense herpesvirus infection. In striking contrast, our knowledge about the physiologic relevance of cytoplasmic RNA sensors of the RIG-I-like receptor (RLR) family in the detection of herpesviruses, such as herpes simplex virus type 1 (HSV-1), is rudimentary. The proposed study builds on a recent discovery by the Gack laboratory that RIG-I plays a crucial role in the detection and restriction of HSV-1 infection, where RIG-I and intracellular DNA sensors act in a temporal manner. Furthermore, we identified that during HSV-1 infection the host-derived 5S ribosomal RNA pseudogene transcript 141 (RNA5SP141) activates RIG-I and elicits cytokine-mediated antiviral innate immune defense. In collaborative work we recently identified that ablated RNA5SP141 gene expression is associated with severe HSV-1 encephalitis in humans. Furthermore, we discovered that RNA5SP141 is exported from infected cells via exosomes and taken up by uninfected cells, where it triggered RIG-I signaling. Using a coordinated series of molecular, biochemical, and cell biological approaches combined with CRISPR-gene editing techniques, we will define the role of RNA5SP141 gene expression in the antiviral innate immune response to HSV-1 infection and in HSV-1-associated disease (AIM 1). Furthermore, we will elucidate the molecular mechanism(s) underlying RNA5SP141 exosomal loading (AIM 2), and also determine the physiologic relevance of exosomal delivery of RNA5SP141 in host immunity to HSV-1 infection (AIM 3). Our studies will provide a molecular understanding of innate immune detection of herpesvirus infection, which may provide the foundation for new therapeutic approaches or guide the design of novel adjuvants.