Conserved Immune Response to Sensing Cytosolic DNA

对感测胞质 DNA 的保守免疫反应

• 批准号: 8889860
• 负责人: Alan Gabriel Goodman
• 金额: $ 24.9万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8889860
• 关键词: Address; Affect; American; Animal Model; Animals; Autoimmune Diseases; Autoimmune Process; Autoimmunity; Award; Bacterial Infections; Binding; Biochemical; Biological Models; Biomedical Engineering; Birth; Cardiovascular system; Cells; Chronic; Collaborations; Complement; Computing Methodologies; DNA; Data; Disease; Double-Stranded RNA; Drosophila Proteins; Drosophila genus; Drosophila melanogaster; Environment; Equipment; Event; Exhibits; Exposure to; Faculty; Fat Body; Gene Activation; Gene Expression; Genes; Genome; Genomic Library; Genomics; Goals; Hemolymph; Homologous Gene; Host Defense; Human; Immune; Immune response; Immune system; Immunology; Infection; Inflammatory; Interferon-beta; Interferons; Invaded; Knock-out; Knowledge; Life Cycle Stages; Ligands; Malignant Neoplasms; Mammals; Maps; Mentors; Mentorship; Microbiology; Microscopy; Modeling; Molecular; Molecular Biology; Natural Immunity; Nerve Degeneration; Nuclear Translocation; Nucleic Acid Binding; Nucleic Acids; Organism; Outcome; Pathogen detection; Pathology; Pathway interactions; Phase; Postdoctoral Fellow; Process; Proteins; RNA Interference; Reaction; Receptor Activation; Receptor Signaling; Research; Research Personnel; Role; Signal Pathway; Signal Transduction; Site; Specificity; Surface; System; Techniques; Testing; Therapeutic Intervention; Training; Transgenic Organisms; Universities; Viral; Virus; Virus Diseases; Work; abstracting; antimicrobial; antimicrobial peptide; base; bone; career; design; fly; gene function; grasp; human disease; improved; insight; loss of function; microbial; mutant; next generation sequencing; novel; pathogen; peptidoglycan recognition protein; receptor; response; sensor; signature molecule; therapeutic development; therapeutic target; therapy development; transcription factor; vaccine development; viral RNA; virology

Conserved Immune Response to Sensing Cytosolic DNA Project Summary/Abstract: Candidate: Throughout my scientific career I have been involved in the study of host-pathogen interactions, with an emphasis on bioengineering, microbiology, immunology, and vaccine development. Training in virology, molecular biology, and computational methods has helped me answer a wide variety of scientific questions. The project proposed here is designed to bridge my transition from a postdoctoral fellow to an independent investigator in an academic setting to continue my pursuit of using creative approaches to high- impact, adventurous research to understand the host response to pathogenic infection. Research: Innate immunity refers to the body's initial response to curb infection upon exposure to invading organisms. While the detection of pathogen-associated molecules is an ancient form of host defense, if dysfunctional, it can cause autoimmune disease, which affects over 20 million Americans. The innate immune response is the first line of defense to microbial infection, and it is initiated through the activation of receptors recognizing conserved molecules that are signature of pathogenic infection. Recently, STING (STimulator of INterferon Genes), an intracellular sensor of cytosolic DNA was discovered. STING is critical to the innate immune response during viral and bacterial infection, yet animals exhibiting STING hyperactivation at birth display inflammatory autoimmune disease. While the downstream signaling events occurring after STING activation are well understood, little is known about the mechanisms responsible for STING activation. To address this question, I propose an orthogonal approach utilizing Drosophila melanogaster to investigate STING function and the molecules that stimulate it. This approach will complement the ongoing studies of STING in a mammalian system. In Aim 1, we will determine how human STING and Drosophila STING (dSTING) are similar, providing evidence that dSTING is the bone fide ancestor of human STING. We will assess the capacity of dSTING to bind nucleic acids and characterize the domains of dSTING to determine which are critical for nucleic acid binding and intracellular localization. In Aim 2, we will create a dSTING knockout fly to study the role of dSTING in the innate immune response to infection. Gene expression analyses will be used to functionally determine how the loss of dSTING contributes to an impaired innate immune response, as compared to that of a genomic rescue fly. Aim 3 will utilize next-generation sequencing to identify the nucleic acids that bind to dSTING during microbial infection, providing insight to the specificity of the ligand for STING, a site for potential therapeutic intervention. Together, these studies using the genetically malleable Drosophila model system will improve our understanding of STING function through the extrapolation of the results into the mammalian system for further experimentation. The information gained in this study will have broad-ranging impacts in innate and autoimmunity towards to the development of therapeutics to treat microbial infection and autoimmune disorders. Environment: This project complements the ongoing research on innate immunity of my proposed mentor, Dr. Glen Barber. The work will also setup new collaborations with my co-mentor, Dr. Grace Zhai. Both mentors will provide invaluable mentorship throughout the K99 phase of the award, giving me the training necessary for the rise to independence. The University of Miami has superb facilities, equipment, and outstanding faculty who study microbiology and immunology, through the use of exciting techniques and animal models.

保守的免疫反应对感应胞质DNA 项目摘要/摘要： 候选人：在我的整个科学生涯中，我都参与了宿主病原体相互作用的研究， 重点是生物工程，微生物学，免疫学和疫苗开发。培训 病毒学，分子生物学和计算方法帮助我回答了各种各样的科学 问题。这里提出的项目旨在弥合我从博士后研究员到一个 在学术环境中的独立调查员继续我追求使用创造性的方法来进行高级 影响，冒险的研究以了解宿主对致病感染的反应。 研究：先天免疫是指暴露于入侵时人体对遏制感染的最初反应 有机体。尽管病原体相关分子的检测是一种古老的宿主防御形式，但如果 功能失调，可能导致自身免疫性疾病，这会影响超过2000万美国人。先天免疫 反应是对微生物感染的第一道防线，它是通过受体激活引发的 识别具有致病感染的保守分子。最近，Sting（刺激器 干扰素基因），发现了细胞内DNA的细胞内传感器。刺对先天至关重要 病毒和细菌感染期间的免疫反应，但出生时表现出刺激性的动物 表现出炎症自身免疫性疾病。而下游信号事件发生在刺痛之后 激活众所周知，对负责刺激激活的机制知之甚少。到 解决了这个问题，我提出了一种使用果蝇Melanogaster的正交方法 研究刺激功能和刺激它的分子。这种方法将补充 正在进行的哺乳动物系统中刺的研究。在AIM 1中，我们将确定人类的刺痛和 果蝇刺（DSTING）是相似的，提供了dsting是人类的骨头祖先的证据 蜇。我们将评估DSTING结合核酸的能力并表征DSTING的结构域 确定哪些对于核酸结合和细胞内定位至关重要。在AIM 2中，我们将创建一个 DSTING敲除飞行以研究DSTIN在先天免疫反应中的作用。基因表达 分析将用于在功能上确定DSTIN的损失如何有助于先天性受损 与基因组救援飞行相比，免疫反应。 AIM 3将使用下一代测序 鉴定在微生物感染过程中与DSTIN结合的核酸，从而洞悉 刺痛的配体，这是潜在治疗干预的部位。一起，这些研究使用了基因 可延展的果蝇模型系统将通过 将结果推断到哺乳动物系统中进行进一步实验。获得的信息 这项研究将对先天性和自身免疫的影响广泛，以发展 治疗微生物感染和自身免疫性疾病的治疗剂。 环境：该项目补充了我提议的导师博士对先天免疫的持续研究。 格伦理发师。这项工作还将与我的同事格蕾丝·盖（Grace Zhai）博士进行新的合作。两位导师 将在整个奖项的K99阶段提供宝贵的指导，为我提供必要的培训 崛起到独立性。迈阿密大学拥有出色的设施，设备和杰出教师 通过使用激动人心的技术和动物模型来研究微生物学和免疫学。