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

破译宿主-病原体相互作用和先天免疫激活期间的转录后基因调控网络

• 批准号: 10624222
• 负责人: Manuel Ascano
• 金额: $ 39.63万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-15 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10624222
• 关键词: Antiviral Response; Autoimmune Diseases; Biological; Biology; Cell physiology; Cells; Critical Pathways; DNA; DNA Damage; DNA-Binding Proteins; Detection; Development; Disease; Environment; Funding; Gene Expression; Genetic Transcription; Goals; Health; Human; Immune; Immune response; Immunologic Surveillance; Immunologics; Infection; Innate Immune Response; Interferons; Investigation; Knowledge; Laboratories; Mission; Modeling; Natural Immunity; Nucleic Acids; Pathogenicity; Pathway interactions; Post-Transcriptional Regulation; Protein Biochemistry; Protein Family; RNA; RNA-Binding Proteins; Regulation; Regulator Genes; Research; Role; Shapes; Stimulator of Interferon Genes; Stimulus; Transcript; Transcriptional Activation; Viral; Viral Genome; Virulence; Virus; Virus Diseases; Virus Replication; cytokine; fitness; gene regulatory network; immune activation; immunogenic; immunoregulation; inhibitor; interest; novel; pathogen; posttranscriptional; prevent; programs; response; sensor; small molecule; tumor

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.

项目总结 我的实验室的长期任务是破译将核酸视为危险的关键途径， 以及促进后续细胞反应的转录后基因调控组件- 目标是这样的知识大大有助于我们对先天的科学理解 免疫在人类健康和疾病中的作用。对先天免疫的研究传统上集中在 了解最终调节中心组转录表达的启动触发因素 干扰素和细胞因子。对转录后基因调控知之甚少。 层，其作用是在RNA水平上优化先天免疫激活-塑造基因表达以 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。考虑到许多方面，这在我们的理解上仍然是一个巨大的差距 宿主-病原体相互作用的核心是检测和抑制外来核酸， 尤其是来自病毒的。RNA结合蛋白(RBPs)可以预先编程细胞对 免疫原性刺激，以及在抗病毒反应中的基本因素。对于这些利益， 与我们概述的R35资助期第一周期的目标一致，我们做出了大量的 我们在以下方面的研究进展：1)必需的和免疫相关的限制性商业惯例的特征 包括ELAVL1和YTHDF蛋白家族，2)广泛存在的病毒宿主RBP的发现 我们开发VIR-CLAP的相互作用及其对病毒影响的表征 3)一种新的小分子催化抑制剂的鉴定和表征。 CGAS-STING途径，鉴于我们还通过研究 这种初级胞质DNA传感器如何启动对感知到的普遍先天免疫反应 外来核酸。总而言之，我的实验室在RNA/DNA结合蛋白方面的专业知识有所增长 生物化学和经济规模生物学旨在直接表征宿主限制性商业惯例和 CGAS途径在赋予先天免疫中的组成部分。对于这次研究更新，我们的目标是 在我们既定的研究计划中，通过开展以下工作来扩大我们的理解 主要的生物学问题： 1)非规范的细胞型限制性商业惯例如何调节和维持宿主基因的表达，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 在早期感染期间不同细胞内环境中的病毒？ 3)调节和促进非典型cGAS依赖途径的新成分是什么？ 促进先天免疫转录激活和程序性DNA损伤。

项目成果

Viral crosslinking and solid-phase purification enables discovery of ribonucleoprotein complexes on incoming RNA virus genomes.

• DOI: 10.1038/s41596-020-00429-6
• 发表时间: 2021-01
• 期刊: Nature protocols
• 影响因子: 14.8
• 作者: Kim B;Arcos S;Rothamel K;Ascano M
• 通讯作者: Ascano M

Small molecule inhibition of cGAS reduces interferon expression in primary macrophages from autoimmune mice.

• DOI: 10.1038/s41467-017-00833-9
• 发表时间: 2017-09-29
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Vincent J;Adura C;Gao P;Luz A;Lama L;Asano Y;Okamoto R;Imaeda T;Aida J;Rothamel K;Gogakos T;Steinberg J;Reasoner S;Aso K;Tuschl T;Patel DJ;Glickman JF;Ascano M
• 通讯作者: Ascano M