解码新生隐球菌赖氨酸乙酰化影响真菌毒性的分子机制和调控网络

病原真菌对人类造成了极大的威胁，每年造成超过200万人死亡。新生隐球菌在病原性真菌引起的死亡人数中居首位，每年导致65万人死亡。对于其致病机制的深入了解是推动药物靶点研发的关键。申请人前期研究发现，乙酰化抑制剂可以有效抑制真菌毒性因子的表达，并且可以大幅度降低宿主中的菌落数量。然而，目前对乙酰化调控隐球菌毒性的分子机制一无所知。本项目以赖氨酸乙酰化(Kac)参与调控新生隐球菌毒性的分子机制为研究方向。首先通过乙酰化蛋白组学方法，构建新生隐球菌全蛋白赖氨酸乙酰化位点图谱，分析Kac调控毒性的可能途径。并以核心毒性因子做为切入点解析Kac修饰决定毒性发挥的关键分子机制。其次，通过系统筛选新生隐球菌脱乙酰化基酶敲除菌株，找出调控毒性的关键性基酶，利用组学、分子生物学、生物信息学等手段，深度解码Kac调控毒性的分子机制。本项目以一个崭新的角度理解真菌毒性的调控途径，为寻求新的药物靶点提供理论依据。

Fungal pathogens are major threats to human health. Cryptococcus neoformans is the leading cause of human death among all fungal pathogens, leading 650,000 deaths per year globally. While current anti-fungal therapies are ineffective, new drug development is hampered by the lack of knowledge of novel molecular targets in fungal pathogenicity. In this study, we discovered that deacetylase inhibitors are potent anti-fungal agents in both in vitro and in animal infection models. Our data demonstrated that deacetylase inhibitors are able to block the expression of genes of many essential virulence factors and dampen fungal proliferation during the early stage of pulmonary infection. However, the molecular mechanism of the effect of deacetylase inhibitors in fungal virulence remains unknown. In our study, we propose to systematically analyze the correlation between post-translational protein acetylation and Cryptococcus neoformans pathogenicity, underlining the importance of acetylation as a novel modulator of fungal virulence. To decipher our hypothesis, we employed acetylome analysis and generated the first database of acetylation network in Cryptococcus neoformans. We detected over 1400 acetylated proteins and more than 3500 acetylated lysine sites. Using bioinformatic analysis, we have identified novel fungal virulence factors, whose activities are regulated by acetylated lysine residues. On the other hand, we generated and screened the deacetylase knockout library, identified two important deacetylases that positively regulate Cryptococcus virulence in the host. We will employ proteomics, acetylomics, molecular biology, and bioinformatic analysis to decipher their functions in modulating fungal virulence. Our study describes a novel regulation of virulence in Cryptococcus neoformans, and provides potential drug targets for anti-fungal therapy development.