Huang Jiahui, Zhao Xuan, Zang Xuelei, Jin Ziyi, Zhang Xueqing, Huang Yemei, Zhang Liye, Xue Xinying, Zhang Ping
Key Laboratory of Cell Proliferation and Regulation Biology, Ministry of Education, College of Life Sciences, Beijing Normal University, Beijing 100875, China.
Department of Respiratory and Critical Care, Emergency and Critical Care Medical Center, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China.
Med Mycol. 2025 Jul 2;63(7). doi: 10.1093/mmy/myaf061.
Cryptococcus gattii is a fungal pathogen that poses significant threats to human health, affecting both immunocompromised and immunocompetent individuals. Treatment of C. gattii infections typically involves the use of antifungal agents, such as azoles. However, the increasing emergence of antifungal resistance in C. gattii is a growing concern, highlighting the critical need for novel therapeutic strategies. In our previous study, we identified a mitochondrial ATP-binding cassette (ABC) transporter, Atm1, as potentially involved in antifungal resistance in C. gattii through transcriptome sequencing, but its function remains unclear and requires additional confirmation and investigation. In this study, we developed a "suicide" clustered regularlyinterspaced short palindromic repeats-CRISPR-associated protein 9 system in C. gattii, based on the system used in C. neoformans, and successfully validated its functionality by targeting the ADE2 gene. We subsequently generated C. gattii mutants lacking ATM1 and assessed their growth under various stress conditions. Our data suggest that Atm1 is involved in the iron-sulfur cluster biosynthesis process. Besides, disruption of ATM1 resulted in various growth impairments, including reduced stress tolerance, impaired capsule formation, and diminished virulence. Importantly, we observed compromised antifungal drug resistance in the atm1∆ mutant and performed RNA sequencing-based transcriptome analysis and gene ontology analysis with and without antifungal treatment for further investigation. In conclusion, our findings indicate that ATM1 plays a role in iron homeostasis and is critical for antifungal resistance in C. gattii, offering new insights into potential drug development strategies for the clinical treatment of cryptococcosis.
加氏隐球菌是一种对人类健康构成重大威胁的真菌病原体，可影响免疫功能低下和免疫功能正常的个体。加氏隐球菌感染的治疗通常涉及使用抗真菌药物，如唑类。然而，加氏隐球菌中抗真菌耐药性的不断出现日益受到关注，凸显了对新型治疗策略的迫切需求。在我们之前的研究中，通过转录组测序，我们鉴定出一种线粒体ATP结合盒（ABC）转运蛋白Atm1可能与加氏隐球菌的抗真菌耐药性有关，但其功能仍不清楚，需要进一步证实和研究。在本研究中，我们基于新生隐球菌中使用的系统，在加氏隐球菌中开发了一种“自杀性”成簇规律间隔短回文重复序列-CRISPR相关蛋白9系统，并通过靶向ADE2基因成功验证了其功能。随后，我们生成了缺乏ATM1的加氏隐球菌突变体，并评估了它们在各种应激条件下的生长情况。我们的数据表明，Atm1参与铁硫簇生物合成过程。此外，ATM1的破坏导致了各种生长缺陷，包括应激耐受性降低、荚膜形成受损和毒力减弱。重要的是，我们观察到atm1∆突变体的抗真菌药物耐药性受损，并对有无抗真菌治疗的情况进行了基于RNA测序的转录组分析和基因本体分析，以进行进一步研究。总之，我们的研究结果表明，ATM1在铁稳态中起作用，对加氏隐球菌的抗真菌耐药性至关重要，为隐球菌病临床治疗的潜在药物开发策略提供了新的见解。
