Effectors of tissue invasion in Aspergillus fumigatus, the major fungal pathogen of human lungs

侵袭性曲霉病是一种经常致命的真菌性肺部疾病，自40年前出现以来，已经夺去了800多万人的生命。ia相关死亡率高得令人无法接受（50- 90%），通常是由于真菌侵袭部位出血导致呼吸衰竭。鉴于抗入侵疗法具有挽救生命的潜力，人们对识别组织入侵的效应物给予了很高的重视，可能会针对这些效应物设计药物和疫苗。然而，对组织侵袭表型和分泌真菌产物之间因果关系的探索一再失败，可能是由于在效应库中可能存在功能冗余，以及烟曲霉遗传方法的不足，无法驱动分泌真菌产物的高通量分析。利用小鼠IA模型、体外感染培养的人肺细胞、功能基因组学和高通量烟熏假单胞菌基因缺失方法，我们首次发现了42个组织入侵效应物（eTINs），它们控制人肺细胞的溶解性死亡。现在，我们将结合最先进的遗传、单细胞成像和效应诱捕方法与我们已建立的方法工作流程，最终定义侵袭性烟熏假单胞菌分泌组，并描述单个效应体的进入模式和/或裂解活性。这项工作将首次揭示组织侵袭性IA病理的分子基础，并确定抗侵袭治疗和新型疫苗和诊断方法可设计的基因组靶点队列。

The often-fatal fungal lung disease invasive aspergillosis (IA) has claimed more than 8 million human lives since its emergence ~ 40 years ago. IA-related mortality is unacceptably high (50- 90%) and usually results from respiratory failure due to haemorrhage occurring at sites of fungal invasion. Given the life-saving potential of anti-invasion therapies, a high premium has been placed upon identifying effectors of tissue invasion, against which drugs and vaccines might be engineered. However, the quest for causal associations between tissue-invasive phenotypes and secreted fungal products has repeatedly failed, presumably due to likely functional redundancy amongst the effector repertoire and insufficiency of genetic approaches in A. fumigatus with which to drive high throughput analyses of secreted fungal products. Using mouse models of IA, in vitro infections of cultured human lung cells, functional genomics and a high throughput A. fumigatus gene deletion methodology we have discovered a first-in-field cohort of 42 effectors of tissue invasion (eTINs) which govern lytic death of human lung cells. We will now combine state-of-the-art genetic, single cell imaging and effector-trap approaches with our established methodological workflow to conclusively define the invasive A. fumigatus secretome and to characterise the modes of entry and/or lytic activity of the individual effectors. This work will reveal for the first time the molecular basis of tissue invasive IA pathologies and define the genomic cohort of targets against which anti-invasion therapies and novel vaccines and diagnostics can be designed.