干扰曲霉凋亡抑制因子Bir1及其重组蛋白对侵袭性曲霉病的影响

The mortality of Invasive aspergillosis (IA) is up to 90%, and effective prevention and treatment of this disease are unsatisfactory. Inhibitor of apoptotic proteins is a kind of important cell apoptosis resistant factor, widely exists in the virus, bacteria, yeast, insects and mammals. Knockout the inhibitor of apoptotic protein Bir1 of Saccharomyces cerevisiae makes it highly sensitive to the external environment of oxidative stress. The amount intracellular ROS was also increased, and which could be induced apoptosis. Through bioinformatics methods, a protein from the genome of Aspergillus fumigatus and Aspergillus flavus were found highly homologous with S. cerevisiae Bir1, respectively. By knockout Bir1 in A. fumigatus and A. flavus and using IA model with mice, the role of Bir1 in the growth, apoptosis, responses to ROS, antifungal drugs susceptibility and pathogenicity were investigated. In order to provide theoretical and experimental basis of novel therapeutic strategy, the preventive effect of recombinant protein of Bir1 in IA were also investigated.

侵袭性曲霉病(IA)病死率高达90%，且防治效果不佳。凋亡抑制因子是一类重要的抗细胞凋亡因子，广泛存在于病毒、细菌、酵母、昆虫和哺乳动物中。酿酒酵母凋亡抑制因子Bir1敲除后使其对外界环境的氧化压力高度敏感。细胞内ROS的水平增加，且可诱导凋亡现象。通过生物信息学方法从烟曲霉和黄曲霉基因组中找出与酿酒酵母Bir1高度同源的烟曲霉和黄曲霉蛋白。本项目通过基因敲除技术及IA 小鼠模型研究Bir1对烟曲霉和黄曲霉生长、凋亡、对ROS的反应、抗真菌药物敏感性以及致病力的影响。并研究Bir1重组蛋白对IA的预防性作用，为提供新的治疗策略提供理论与实验基础。

侵袭性曲霉病是引起免疫低下患者死亡的重要病因之一；现有抗真菌药对侵袭性曲霉病疗效十分不理想，加上唑类耐药烟曲霉的出现使得侵袭性曲霉病的治疗更受限制侵袭性曲霉病是引起免疫低下患者死亡的重要病因之一；现有抗真菌药对侵袭性曲霉病疗效十分不理想，加上唑类耐药烟曲霉的出现使得侵袭性曲霉病的治疗更受限制。我们通过对烟曲霉凋亡抑制因子蛋白BIR1的敲除，发现BIR1的缺陷导致烟曲霉生长延缓，分生孢子头发育缺陷、产孢减少，且对侵袭性曲霉病一线治疗药物伏力康唑敏感性明显增高。同时我们利用微量液基稀释法检测了TOR通路抑制剂INK128、Calcineurin抑制剂他克莫司分别联合唑类药物对曲霉的抗菌作用，发现均有不同程度的协调抗曲霉作用。为将来的临床应用奠定了基础。