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

描述（由申请人提供）：抗菌素耐药性的传播速度迅速超过了新型抗生素的开发速度，对人类健康和福祉构成了迫在眉睫的危险。避免这一危机的一个潜在方法是识别能够刺激先天免疫系统的化合物，从而增加宿主对微生物的清除。此外，这些化合物可能被证明是微生物适应性遗传过程的一个更困难的目标，因为它们破坏宿主-病原体相互作用的平衡，而不是简单地杀死病原体或阻止其复制。在这项应用中，我们将利用一种简单的模式生物秀丽隐杆线虫，它具有几个实验优势：对遗传筛选的适应性，包括正向、反向和化学，对生化分析的适应性，以及与人类相似的进化保守的先天免疫途径。我开发了一种新的液体感染试验，并对其进行了部分表征，并用它来进行高通量筛选，以识别免疫刺激化合物。在这个项目中，将使用转录报告基因来测试减轻感染的化合物激活几种关键免疫和应激途径的能力。这些化合物将用于鉴定秀丽隐杆线虫免疫反应途径的新成分，并确定选定化合物的靶点。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式生物中进一步测试，潜在地为人类治疗的发展确定新的靶点。

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.