Decoding the pathogenic interactome of Aspergillus fumigatus

描述（由申请人提供）：烟曲霉是一种严重的医学威胁，是导致人类疾病的最严重丝状霉菌，总体报告死亡率为60%。随着免疫功能低下的易感人群的不断增加，以及抗真菌耐药真菌分离株的发病率不断上升，迫切需要新的诊断和治疗方法。鉴定适合诊断或治疗开发的真菌和宿主分子是本提案的主要目标。从A.与寄主的相互作用，共发现30株A.烟曲霉转录因子（TF）对疾病发展中重要的两种真菌形态至关重要：初始感染所需的孢子和菌丝，真菌在体内侵入性生长的形式。因此，目的1集中于鉴定孢子军备和真菌/宿主串扰在最初的主机遇到介导的孢子特异性TF。目的2鉴定真菌TF和宿主分子，使侵入性菌丝在体内生长。与目标1和2同时，目标3将开发最先进的微流体平台，首次允许， 筛选1000个A.烟曲霉突变体的入侵生长特性和宿主激活A.烟毒蛋白后一个目标对于曲霉菌研究界至关重要 由于产生A相对容易。fumigatus突变体和管道的完整A.烟曲霉基因组敲除文库（10，000个基因）;筛选如此多的突变体使得动物研究成为评估烟曲霉基因组敲除文库的主要障碍。烟曲霉毒力因子事实上，曲霉菌宿主相互作用的研究受到缺乏可用的鲁棒性的限制。 这些平台（i）能够可视化密切的真菌/宿主相互作用，（ii）允许快速评估毒力性状或（iii）评估真菌和宿主信号分子的作用，这些分子难以获得足够大的数量进行宏观研究。Aim 3将提供这些平台，以帮助研究界满足曲霉基因组时代的需求。

DESCRIPTION (provided by applicant): Aspergillus fumigatus is a serious medical menace and ranks as the most severe filamentous mold causing human disease with 60% overall reported fatality. The increasing population of susceptible immunocompromised individuals coupled with the rising incidence of antifungal resistant fungal isolates underscore the need for new diagnostic and therapeutic modalities. Identification of fungal and host molecules suitable for diagnostic or therapy development is the major goal of this proposal. Compiling data from genomic studies of A. fumigatus/host interactions, we have uncovered 30 A. fumigatus transcription factors (TFs) critical for the two fungal morphologies important in disease development: spores required for initial infection and hyphae, the form of the fungus growing invasively in vivo. Aim 1 thus focuses on identification of spore armaments and fungal/host crosstalk in initial host encounters mediated by spore specific TFs. Aim 2 characterizes the fungal TFs and host molecules enabling invasive hyphal growth in vivo. Simultaneously to Aims 1 and 2, Aim 3 will develop state-of-the-art microfluidic platforms that allow, for the first time, screening of 1000s of A. fumigatus mutants for invasive growth properties and host activation of A. fumigatus virulence proteins. This latter aim is critical for the Aspergillus research community due to the relative ease of generating A. fumigatus mutants and the pipeline availability of a complete A. fumigatus genome knockout library (10,000 genes); screening of this many mutants renders animal studies a major roadblock in assessment of A. fumigatus virulence factors. Indeed, the study of Aspergillus host interactions has been limited by the lack of available robust platforms that (i) enable visualization of intimate fungal/host interactions, (ii) allow for rapid assessment of virulence traits or (iii) evaluate the role of fungal and host signaling molecules that are difficult to obtain in large enough quantities for macrostudies. Aim 3 will provide these platforms to help the research community meet the needs of the Aspergillus genomic era.