Investigating the function of macrophages in the efficacy of anti-fungal drugs in larval zebrafish

项目概要/摘要 真菌病原体在免疫功能低下的患者中引起危及生命的疾病， 全世界每年有100万人受到影响。临床上用于治疗的抗真菌药物 病人是无效的，即使这些药物对培养皿中的真菌效果很好。的 拟议研究的总体目标是提高这些药物在体内的疗效。 活的有机体斑马鱼幼鱼是解决这一问题的理想宿主。优秀的现场 虽然有成像和遗传工具，但斑马鱼和人类的免疫系统在很大程度上 斑马鱼中保守的和真菌感染模型再现了人类的发病机制。初步 数据表明，抗真菌药物伏立康唑和体内巨噬细胞之间的协同作用 增加对真菌病原体烟曲霉的杀灭。我建议识别基因， 巨噬细胞中受唑处理调节并促进唑介导 真菌杀灭首先，我将集中在已知的细胞生物学途径参与病原体识别 和吞噬体杀伤，包括病原体识别受体（PRR）途径，活性氧 物种（ROS）的产生和吞噬体酸化。然后，我将使用一个基于无偏RNAseq的 方法来确定未知的基因是由唑治疗调制。这两个都有 这些方法在未来可以作为辅助治疗的目标，以提高抗真菌治疗的疗效。 感染的宿主总之，这项提议的结果将确定免疫治疗的新靶点， 治疗性辅助治疗，以增加抗真菌治疗患者的疗效。

Project Summary/Abstract Fungal pathogens cause life-threatening disease in immuno-compromised patients, with more than 2 million people affected world-wide each year. Anti-fungal drugs that are used in the clinic to treat patients are ineffective, even though these drugs work well against fungi in a petri dish. The overarching goal of the proposed research is to increase the efficacy of these drugs inside of living organisms. The larval zebrafish is an ideal host in which to tackle this problem. Excellent live imaging and genetic tools are available, the immune systems of zebrafish and humans are largely conserved, and fungal infection models in zebrafish recapitulate pathogenesis in humans. Preliminary data indicates that synergy between the anti-fungal drug voriconazole and macrophages in vivo increases killing of the fungal pathogen Aspergillus fumigatus. I propose to identify genes and pathways in macrophages that are modulated by azole treatment and that promote azole-mediated fungal killing. First, I will focus on known cell biological pathways involved in pathogen recognition and phagosomal killing, including pathogen recognition receptor (PRR) pathways, reactive oxygen species (ROS) generation, and phagosomal acidification. Then, I will use an unbiased RNAseq-based approach to identify unknown genes that are modulated by azole treatment. Hits from both of these approaches could be targeted in the future for adjuvant therapy to boost anti-fungal efficacy in infected hosts. Altogether, results from this proposal will identify new targets for immuno- therapeutic adjuvant therapy to increase the efficacy of anti-fungal treatment in patients.