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

基本信息

批准号：
8176709
负责人：
CHRISTIAN W SCHINDLER
金额：
$ 24.01万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-06-07 至 2013-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8176709
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

项目摘要

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. PUBLIC HEALTH RELEVANCE: Type I Interferons (IFN-Is), known for their antiviral activity, also direct an effective innate response towards facultative intracellular bacteria, like Francisella tularensis and Legionella pneumophila. The proposed studies explore the mechanism by which c-diGMP, an important L. pneumophila regulatory molecule, stimulates NF-:B activation and IFN-I expression. These studies focus on identifying a new innate immune response pathway, which has important implications on antibacterial therapy and vaccine development.

描述（由申请人提供）：由细菌引起的传染病，尤其是抗生素耐药病原体，已成为美国医疗保健系统的越来越多的负担，医院相关感染的直接医疗费用每年估计在300-45亿美元之间。此外，新的抗生素的发展无法与更具耐药性细菌的快速出现保持同步。幸运的是，已经描述了与宿主共同发展的越来越多的先天的“传感器和响应系统”。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”。这些传感器启动了众所周知的促炎信号传导级联反应，这些级联反应以TNF-1和干扰素（IFNS）等免疫效应子的表达。这些传感器和响应系统的表征为发展同样复杂的新治疗策略提供了重要的机会。为了表征将有效反应对肺炎军团菌的有效反应的天生反应，我们确定了一种新型的细菌调节分子，3'，5'-循环二甘氨酸酯（C-DIGMP）在刺激自由分泌IFN的表达中起着有效抑制细菌生长的表达中起着重要作用。最近也报道了对源自李斯特菌的单核细胞增生李斯特菌的类似反应，最近也报道了3'，5'-循环二氨基二二酰二氨基二二酰二氨基二氨核酸二氨基丁酸（C-DIAMP）。我们提出了生化和遗传方法，以表征专门响应C-DIGMP的哺乳动物传感器系统的成分。为此，我们开发了生物素修饰的C-DIGMP和C-DIGMP报告基细胞系。我们预计我们识别或密切相关的同源物的传感器组件将指导对c-DIAMP的响应。具体而言，我们提出：1。利用亲和力色谱法鉴定C-DIGMP的哺乳动物受体。 2。利用遗传筛选来识别直接对C-DIGMP炎症反应的哺乳动物传感器系统的成分。公共卫生相关性：以抗病毒活性而闻名的I型干扰素（IFN-IS）也将有效的先天反应对辅助性细胞内细菌（如Francisella toarlensis和pneumophila）产生。提出的研究探讨了重要的肺炎乳杆菌调节分子C-DIGMP刺激NF-：B激活和IFN-I表达的机制。这些研究着重于确定一种新的先天免疫反应途径，该途径对抗菌治疗和疫苗发育具有重要意义。