Deciphering Post-transcriptional Gene Regulatory Networks During Periods of Host-Pathogen Interaction and Innate Immune Activation

项目摘要 我实验室的长期使命是破译将核酸视为危险的关键途径， 以及促进随后细胞反应的转录后基因调控成分- 目标是这些知识大大有助于我们对先天性的科学理解 免疫力对人类健康和疾病的影响。对先天免疫的研究传统上集中在 理解最终调节中枢集转录表达的起始触发因素 干扰素和细胞因子。关于转录后基因调控的知之甚少， 层，其作用是在RNA水平上改善先天免疫激活-塑造基因表达， 允许强大但有限的宿主反应，同时防止异常或病原体- 相关基因表达。这仍然是我们认识上的一个显著差距，因为许多方面 宿主-病原体相互作用的核心是检测和抑制外源核酸， 特别是病毒。RNA结合蛋白（RBP）可以预先编程细胞对RNA的敏感性。 免疫原性刺激，以及在抗病毒反应的基本因素。为了这些利益， 根据我们R35资助期第一周期的目标，我们取得了实质性进展， 我们在以下方面的研究进展：1）基本和免疫相关RBP的表征 包括ELAVL 1和YTHDF蛋白家族，2）发现广泛的病毒宿主RBP 通过我们开发的VIR-CLASP和表征它们对病毒的影响， 复制，和3）一种新的小分子催化抑制剂的鉴定和表征， cGAS-STING途径，因为我们还通过检查 这种主要的胞质DNA传感器如何启动普遍的先天免疫反应， 外源核酸总的来说，我的实验室在RNA/DNA结合蛋白方面的专业知识有所增长， 生物化学和组学规模的生物学，旨在直接表征宿主RBP的基本作用， 在赋予先天免疫中cGAS途径的组分。对于这项研究更新，我们的目标是 通过以下方式扩大我们在既定研究计划中的理解 主要的生物学问题： 1)非典型细胞RBP如何调节和维持宿主基因表达，特别是在病毒感染期间， 感染？ 2)影响相关病毒复制和毒力的独特宿主RBP-病毒基因组相互作用是什么？ 病毒在不同的细胞内环境在早期感染？ 3)有哪些新的组分调节和促进非经典cGAS依赖性途径 促进先天免疫转录激活和程序性DNA损伤。

PROJECT SUMMARY The long-term mission of my lab is to decipher the critical pathways that sense nucleic acids as a danger, and the post-transcriptional gene regulatory components that facilitate the subsequent cellular response - with the goal that such knowledge significantly contributes to our scientific understanding of innate immunity in human health and disease. Research on innate immunity has traditionally focused on understanding the initiating triggers that ultimately modulate the transcriptional expression of a central set of interferons and cytokines. Much less is known about the post-transcriptional gene regulatory layer, which acts to refine innate immune activation at the RNA level – shaping gene expression to allow for a robust but finite host response while simultaneously preventing aberrant or pathogen- associated gene expression. This remains a striking gap in our understanding given that many aspects of host-pathogen interactions have at its core the detection and suppression of foreign nucleic acids, particularly from viruses. RNA-binding proteins (RBPs) can pre-program the sensitivity of cells to immunogenic stimuli, as well as being essential factors in the anti-viral response. Towards these interests, and in line with our outlined goals of the first cycle of our R35 funding period, we made substantial progress in our investigations on: 1)The characterization of essential and immune-relevant RBPs including ELAVL1 and the YTHDF protein family, 2) the discovery of widespread viral-host RBP interactions through our development of VIR-CLASP and characterization of their impact on viral replication, and 3) the identification and characterization of a novel small molecule catalytic inhibitor of the cGAS-STING pathway, given that we also model dynamic post-transcriptional regulation by examining how this primary cytosolic DNA sensor initiates a generalized innate immune response to perceived foreign nucleic acids. Taken together, my laboratory has grown in expertise in RNA/DNA binding protein biochemistry and -omic scale biology aimed directly at characterizing the essential roles of host RBPs and components of the cGAS pathway in conferring innate immunity. For this research renewal, we aim to expand our understanding within our established research program by pursuing the following major biological questions: 1)How do non-canonical cellular RBPs regulate and maintain host gene expression, especially during viral infection? 2)What are the unique host RBP-viral genome interactions that impact replication and virulence of related viruses within different intracellular environments during early infection? 3) What are the novel components that regulate and facilitate a non-canonical cGAS-dependent pathway promoting innate immune transcriptional activation and programmed DNA damage.