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

项目摘要/摘要 在免疫功能低下的患者中，真菌病原体会导致危及生命的疾病，有超过2种 全球每年有数百万人受到影响。临床上用于治疗的抗真菌药物 患者是无效的，即使这些药物对培养皿中的真菌很有效。这个 这项拟议研究的首要目标是提高这些药物在 活着的有机体。斑马鱼幼体是解决这一问题的理想宿主。精彩的现场直播 成像和遗传工具是可用的，斑马鱼和人类的免疫系统在很大程度上 斑马鱼中的保守和真菌感染模型概括了人类的发病机制。初步 数据表明，抗真菌药物伏立康唑与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作用。我建议识别基因和 巨噬细胞中受唑处理调节并促进唑介化的通路 真菌致死。首先，我将重点介绍病原体识别所涉及的已知细胞生物学途径。 和吞噬体杀，包括病原体识别受体(PRR)途径，活性氧 物种(ROS)的生成和吞噬体酸化。然后，我将使用基于RNAseq的无偏见 一种识别受唑类药物调控的未知基因的方法。这两个网站的点击率都是 未来可能有针对性的辅助治疗方法，以提高抗真菌疗效 受感染的宿主。总之，这项提案的结果将确定免疫的新靶点。 治疗辅助治疗，提高患者抗真菌治疗的疗效。

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.