棘白菌素胁迫下光滑念珠菌持留状态形成与发展机制的多组学研究

棘白菌素耐药型光滑念珠菌的出现给临床治疗带来严峻挑战。本课题组在棘白菌素体外杀菌试验中发现，光滑念珠菌在形成稳定耐药前存在持留状态。细菌研究中证实，在高浓度抗菌药物胁迫下持留状态的形成能够提高菌株的存活率和耐药基因突变率，与耐药发展密切相关，但在念珠菌中相关研究匮乏。因此本项目提出重要科学假说，光滑念珠菌在棘白菌素胁迫下形成的持留状态是发展为稳定耐药的重要前提。本项目将1）分析药物作用时间和浓度对持留状态形成的影响；2）从转录组和蛋白质组学角度，联合分析持留菌形成和发展过程中RNA和蛋白水平的动态调控网络，筛选对持留菌形成和发展起关键作用的靶基因，并验证其功能；3）检测我国临床菌株的药物敏感性和持留菌靶基因的表达水平，分析靶基因在不同菌株中的差异。本项目拟通过上述研究，探索棘白菌素胁迫下光滑念珠菌持留状态形成与发展机制，为控制持留状态发生发展、遏制真菌耐药和指导临床合理用药提供新思路。

The emergence of Candida glabrata isolates resistant to echinocandins has proposed great challenges to clinical treatment. In the preliminary in vitro killing assay experiment, we found C. glabrata persister was formed under the stress of echinocandins. It was reported that bacterial persister can increase the number of survival cells and mutation rates under high concentrations of antimicrobial agents, which closely related to resistance development. However, less is known about fungal persister. On this basis, a scientific hypothesis was proposed that the persistence was an important stage before the development of echinocandin resistance. Firstly, our research will analyze the influence from duration of action and concentration of echinocandins on the formation of C. glabrata persistence. Then we will explore the homeostatic molecular mechanism of C. glabrata persistence, based on the multi-omics study including transcriptome and proteome. We will also screen and validate the persister target genes based on expression level of transcription and protein. The antifungal susceptibility and expression level of target genes in clinical invasive C. glabrata isolates will be analyzed. Through overall analysis, this study means to confirm the mechanism of the formation and development of the C. glabrata persister under the stress of echinocandins, which could provide new ideas on controlling C. glabrata persistence formation, reducing clinical resistance and guiding clinical medication.