Utilizing a novel liquid-killing assay to gain insight into C. elegans immunity

利用新型液体杀灭测定来深入了解秀丽隐杆线虫的免疫力

基本信息

批准号：
8311400
负责人：
Natasha Kirienko
金额：
$ 5.22万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-02-01 至 2015-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8311400
关键词：
Agar Animal Model Antibiotic Resistance Antibiotic Therapy Antibiotics Antimicrobial Resistance Bacterial Infections Biochemical Biological Assay Biological Models Biology Caenorhabditis elegans Cell Culture Techniques Chemicals Data Development Drosophila genus Drosophila melanogaster Eligibility Determination Enterococcus faecalis Enterococcus faecium Future Generations Genes Genetic Genetic Epistasis Genetic Processes Genetic Screening Goals Health Human Human Development Immune Immune response Immune system Immunity Immunization Infection Invertebrates Knowledge Laboratories Liquid substance Mammalian Cell Mammals Maps Mediating Methods Microbe Molecular Biology Morbidity - disease rate National Research Service Awards Natural Immunity Nematoda Organism Outcome Pathogenesis Pathway interactions Personal Satisfaction Pharmaceutical Preparations Pseudomonas aeruginosa Reporter Resistance Role Signal Pathway Signal Transduction Societies Streptococcus pneumoniae Stress Testing Therapeutic Translating Vancomycin resistant enterococcus Vertebrates Work antimicrobial arm biological adaptation to stress chemical genetics high throughput screening immune activation insight killings microbial mortality novel pathogen prevent research study response small molecule libraries

项目摘要

DESCRIPTION (provided by applicant): The spread of antimicrobial resistance is quickly outstripping the development of novel antibiotics, suggesting a looming danger for human health and well-being. One potential means for avoiding this crisis is the identification of compounds that stimulate the innate immune system, triggering increased microbial clearance by the host. In addition, these compounds are likely to prove a more difficult target for the adaptive genetic processes of microbes, as they unbalance the host-pathogen interaction, rather than simply killing the pathogen or preventing its replication. For this application, we will utilize a simple model organism, C. elegans, which possesses several experimental advantages: amenability to genetic screens, including forward, reverse, and chemical, adaptability to biochemical assays, and an evolutionarily conserved innate immune pathway similar to those of humans. I developed and partially characterized a novel liquid infection assay with this organism and used it to carry out a high-throughput screen to identify immunostimulatory compounds. For this project, infection-alleviating compounds will be tested for their ability to activate several key immune and stress pathways using transcriptional reporters. The compounds will be used to identify novel components of C. elegans immune response pathways and the targets of selected compounds will be determined. In future work, these compounds can be further tested in a variety of invertebrate and vertebrate model organisms, potentially identifying novel targets for the development of human therapeutics. PUBLIC HEALTH RELEVANCE: The expanding threat of antimicrobial resistance is rapidly outstripping the development of novel antibiotics, leading to a burgeoning menace of untreatable bacterial infections. A novel infection assay with a small worm was used to identify compounds that enhance host survival. Drugs will be tested for immunostimulatory activity, and positive hits will be analyzed to determine their mechanisms of function and will be used to identify novel components of innate immune signaling pathways.

描述（由申请人提供）：抗菌素耐药性的传播速度很快超过了新型抗生素的发展速度，这表明人类健康和福祉面临迫在眉睫的危险。避免这场危机的一种潜在方法是识别刺激先天免疫系统的化合物，从而引发宿主增加微生物清除能力。此外，这些化合物很可能成为微生物适应性遗传过程更困难的目标，因为它们不平衡宿主与病原体的相互作用，而不是简单地杀死病原体或阻止其复制。对于此应用，我们将利用一种简单的模型生物，即秀丽隐杆线虫，它具有多种实验优势：适合遗传筛选，包括正向、反向和化学筛选，对生化测定的适应性，以及类似于那些的进化上保守的先天免疫途径。人类的。我开发并部分表征了这种生物体的新型液体感染测定，并用它进行高通量筛选以鉴定免疫刺激化合物。在这个项目中，将测试减轻感染的化合物使用转录报告基因激活几个关键免疫和应激途径的能力。这些化合物将用于鉴定线虫免疫反应途径的新成分，并将确定所选化合物的靶标。在未来的工作中，这些化合物可以在各种无脊椎动物和脊椎动物模型生物体中进行进一步测试，有可能确定人类治疗药物开发的新靶点。公共卫生相关性：不断扩大的抗菌素耐药性威胁正在迅速超过新型抗生素的发展速度，导致无法治疗的细菌感染的威胁日益严重。一种新型的小蠕虫感染试验被用来鉴定可增强宿主生存的化合物。将测试药物的免疫刺激活性，并对阳性命中进行分析以确定其功能机制，并将用于识别先天免疫信号传导途径的新成分。