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的丝状霉菌，总报告死亡率为60%。对免疫功能敏感的个体数量不断增加，加上耐药真菌分离物的发病率不断上升，凸显了对新的诊断和治疗方式的需要。鉴定适合诊断或治疗发展的真菌和宿主分子是这项提议的主要目标。汇编烟曲霉/宿主相互作用的基因组研究数据，我们发现了30个烟曲霉转录因子(TF)，它们对疾病发展中两种真菌形态至关重要：初始感染所需的孢子和菌丝，这是真菌在体内侵袭性生长的形式。目的1因此重点研究孢子特异性因子介导的初始寄主接触中孢子武器和真菌/寄主串扰的鉴定。目的2表征真菌因子和宿主分子使侵袭性菌丝在体内生长。与目标1和目标2同时，目标3将开发最先进的微流控平台，首次允许， 1000株烟曲霉变异株侵袭生长特性及毒力蛋白宿主激活的筛选。这后一个目标对于曲霉研究界来说是至关重要的 由于产生烟曲霉突变株相对容易，而且有可能获得完整的烟曲霉基因组敲除文库(10,000个基因)；对这许多突变株的筛选使动物研究成为评估烟曲霉毒力因子的主要障碍。事实上，曲霉寄主相互作用的研究由于缺乏可用的稳健数据而受到限制。 这些平台(I)能够可视化真菌/宿主的密切相互作用，(Ii)能够快速评估毒力特征，或者(Iii)评估真菌和宿主信号分子的作用，这些分子很难获得足够多的数量来进行宏观研究。AIM 3将提供这些平台来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.