Probing the Innate Response to 3', 5'-cyclic Diguanylate (c-diGMP)

探索对 3, 5-环二鸟苷酸 (c-diGMP) 的先天反应

• 批准号: 8274633
• 负责人: CHRISTIAN W SCHINDLER
• 金额: $ 20万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-07 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8274633
• 关键词: Affinity Chromatography; Anti-Bacterial Agents; Antibiotic Resistance; Antibiotics; Antiviral Agents; Bacteria; Biochemical; Biochemical Genetics; Biotin; C Type Lectin Receptors; Cell Line; Communicable Diseases; Cyclic Nucleotides; Cytosol; DNA; DNA Polymerase III; Development; Dose; Double-Stranded RNA; Environment; Exclusion; Family; Flagellin; Francisella tularensis; Genetic Screening; Growth; Healthcare Systems; Homologous Gene; Hospital Costs; IRF3 gene; Immune; Immune response; Immune system; Infection; Inflammatory; Inflammatory Response; Interferon Activation; Interferon Type I; Interferons; Interleukin-6; Legionella pneumophila; Libraries; Limb structure; Listeria monocytogenes; Medical; Membrane; Methods; Microbe; Molecular; Nucleic Acids; Nucleotides; Pathway interactions; Pattern; Pattern recognition receptor; Play; Proteins; RNA; Radiolabeled; Reporter; Reporting; Role; Sampling; Signal Transduction; Structure; System; TBK1 gene; TLR3 gene; TLR7 gene; TLR8 gene; TNF gene; Toll-like receptors; Transcription Factor AP-1; Vertebrates; Viral; analog; autocrine; bacterial resistance; chemokine; cytokine; diguanylate cyclase; macrophage; member; microbial; novel; novel therapeutics; pathogen; phosphoric diester hydrolase; radiotracer; receptor; response; sensor; sugar; therapy development; transcription factor; vaccine development

DESCRIPTION (provided by applicant): Infectious diseases caused by bacteria, especially antibiotic resistant pathogens, have become an increasing burden on the US health care system, with direct medical costs for hospital associated infections estimated to range between 30-45 billion dollars annually. Moreover, the development of new antibiotics has been unable to keep pace with the rapid emergence of ever more resistant bacteria. Fortunately, a growing number of sophisticated innate "Sensor and Response Systems" that co-evolved with their hosts have been described. In vertebrates this includes several distinct families of innate sensors that recognize specialized bacterial molecules or "molecular patterns". These sensors initiate well-known pro-inflammatory signaling cascades that culminate in the expression of immune effectors, like TNF-1 and Interferons (IFNs). Characterization of theses Sensor and Responses Systems provides an important opportunity for the development of equally sophisticated new therapeutic strategies. In an effort to characterize the innate responses that direct an effective response towards Legionella pneumophila, we have determined that a novel bacterial regulatory molecule, 3',5'- cyclic diguanylate (c-diGMP) plays an important role in stimulating the expression of autocrine IFNs, which effectively suppress bacterial growth. An analogous response to Listeria monocytogenes derived 3',5'-cyclic diadenylate (c-diAMP), has also recently been reported. We propose biochemical and genetic approaches to characterize the components of the mammalian sensor system that specifically responds to c-diGMP. To this end we have developed a biotin modified c-diGMP and a c-diGMP reporter cell line. We anticipate that the sensor components we identify, or closely related homologs, will direct the response to c- diAMP. Specifically we propose to: 1. Exploit affinity chromatography to identify the mammalian receptor for c-diGMP. 2. Exploit a genetic screen to identify the components of the mammalian sensor system that direct inflammatory response to c-diGMP.

描述（由申请人提供）：由细菌引起的传染病，特别是抗生素耐药病原体，已经成为美国医疗保健系统日益增加的负担，估计每年医院相关感染的直接医疗费用在300 - 450亿美元之间。此外，新抗生素的开发一直无法跟上越来越多的耐药细菌的快速出现。幸运的是，越来越多的复杂的先天“传感器和反应系统”，与他们的主机共同进化已被描述。在脊椎动物中，这包括几个不同的先天传感器家族，它们识别专门的细菌分子或“分子模式”。这些传感器启动众所周知的促炎信号级联，最终导致免疫效应物如TNF-1和干扰素（IFN）的表达。这些传感器和反应系统的表征为开发同样复杂的新治疗策略提供了重要机会。 为了表征针对嗜肺军团菌的有效应答的先天性应答，我们已经确定了一种新的细菌调节分子，3 '，5'-环二鸟苷酸（c-diGMP）在刺激自分泌IFN的表达中起重要作用，其有效地抑制细菌生长。最近也报道了对单核细胞增生李斯特菌衍生的3 '，5'-环二腺苷酸（c-diAMP）的类似应答。我们提出了生物化学和遗传学的方法来表征哺乳动物的传感器系统，专门响应c-diGMP的组件。为此，我们开发了生物素修饰的c-diGMP和c-diGMP报告细胞系。我们预期我们鉴定的传感器组分或密切相关的同系物将指导对c-diAMP的反应。具体而言，我们建议：1。利用亲和色谱法鉴定c-diGMP的哺乳动物受体。 2.利用遗传筛选来识别哺乳动物传感器系统的组件，直接炎症反应的c-diGMP。