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.

对敏感胞浆DNA的保守免疫应答 项目摘要/摘要： 候选人：在我的整个科学生涯中，我一直从事宿主与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 问题。这里提出的项目旨在为我从博士后到博士后的过渡搭建桥梁 在学术环境中担任独立调查员，继续追求使用创造性的方法来 影响，冒险研究，以了解宿主对病原性感染的反应。 研究：先天免疫是指人体在接触入侵时抑制感染的最初反应 有机体。虽然检测病原体相关分子是一种古老的宿主防御形式，但如果 如果功能失调，它会导致自身免疫性疾病，影响超过2000万美国人。先天免疫 反应是抵御微生物感染的第一道防线，它是通过激活受体而启动的。 识别作为病原性感染标志的保守分子。最近，刺痛(刺激物) 干扰素基因)是胞浆DNA的胞内感受器。刺痛对于先天的 病毒和细菌感染时的免疫反应，但动物在出生时表现出刺痛过度 表现为炎症性自身免疫性疾病。而STIN之后发生的下行信令事件 激活是众所周知的，但对刺痛激活的机制知之甚少。至 针对这个问题，我提出了一种利用果蝇的正交化方法来 研究刺的功能和刺激它的分子。这种方法将补充 目前正在对哺乳动物系统中的刺痛进行研究。在目标1中，我们将确定人类如何叮咬和 果蝇的刺(Dstingg)是相似的，这为dstingg是人类的真正祖先提供了证据 刺痛。我们将评估dSTING与核酸结合的能力，并描述dSTING的结构域 以确定哪些对核酸结合和细胞内定位是关键的。在目标2中，我们将创建一个 DSTING基因敲除苍蝇研究DSTING在感染的先天性免疫反应中的作用。基因表达 分析将被用来从功能上确定dSTING的丢失如何导致先天受损 免疫反应，与基因组救援苍蝇的免疫反应相比。AIM 3将利用下一代测序 鉴定在微生物感染过程中与dSTING结合的核酸，为深入了解dSTING的特异性提供依据 刺痛的配基，一个潜在的治疗干预部位。总的来说，这些研究使用了基因上的 可延展的果蝇模型系统将通过 将结果外推到哺乳动物系统中进行进一步实验。中获得的信息 这项研究将对先天免疫和自身免疫的发展产生广泛的影响。 治疗微生物感染和自身免疫性疾病的疗法。 环境：这个项目是对我推荐的导师Dr。 格伦·巴伯。这项工作还将建立与我的共同导师翟志伟博士的新合作。两位导师 将在K99奖项的整个阶段提供宝贵的指导，给我必要的培训 走向独立。迈阿密大学拥有一流的设施、设备和优秀的教职员工 他们研究微生物学和免疫学，通过使用令人兴奋的技术和动物模型。