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

描述(申请人提供)：抗菌素耐药性的传播正在迅速超过新型抗生素的发展，这表明对人类健康和福祉的威胁迫在眉睫。避免这一危机的一个潜在方法是识别能够刺激天然免疫系统的化合物，从而增加宿主对微生物的清除。此外，这些化合物可能会成为微生物适应遗传过程的更难靶标，因为它们破坏了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。对于这一应用，我们将利用一种简单的模式生物，线虫，它具有几个实验优势：对遗传筛选(包括正向、反向和化学)的适应性，对生化分析的适应性，以及进化上保守的类似于人类的先天免疫途径。我用这种细菌开发了一种新的液体感染试验，并对其进行了部分表征，并用它进行了高通量筛选，以鉴定免疫刺激化合物。对于这个项目，缓解感染的化合物将使用转录报告来测试它们激活几个关键的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，并将确定选定化合物的目标。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一步测试，有可能为人类疗法的发展确定新的靶点。 与公共卫生相关：抗菌素耐药性的日益扩大的威胁正在迅速超过新型抗生素的开发，导致无法治疗的细菌感染的威胁迅速增长。一种新的小蠕虫感染试验被用来鉴定能提高宿主存活率的化合物。药物将接受免疫刺激活性测试，阳性HITS将被分析以确定其作用机制，并将用于识别先天免疫信号通路的新成分。

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.