Calcimycin Inhibits Cryptococcus neoformans In Vitro and In Vivo by Targeting the Prp8 Intein Splicing.
Calcimycin Inhibits Cryptococcus neoformans In Vitro and In Vivo by Targeting the Prp8 Intein Splicing.
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DOI：
10.1021/acsinfecdis.2c00137
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发表时间：
2022-09-09
影响因子：
5.3
通讯作者：
Li, Hongmin
中科院分区：
文献类型：
--
作者：
Tharappel, Anil Mathew;Li, Zhong;Zhu, Yan Chun;Wu, Xiangmeng;Chaturvedi, Sudha;Zhang, Qing-Yu;Li, Hongmin
耐药性是疾病治疗中的一个重要问题，包括由新型隐球菌（Cne）和格特隐球菌（Cga）引起的隐球菌病。替代药物靶标对于在耐药性达到关键阶段之前克服耐药性是必要的。内含肽从前蛋白前体的剪接对于许多生物体（包括人类病原体如Cne和Cga）中的内含肽元件的必需蛋白质的活性至关重要。通过高通量筛选，我们鉴定了卡西霉素（CMN）作为有效的Prp 8内含肽剪接抑制剂，其对野生型Cne-H99（Cne-WT或Cne）的最小抑制浓度（MIC）为1.5 μg/mL。相比之下，CMN抑制内含肽较少的突变株（Cne-Mut），MIC高16倍。有趣的是，烟曲霉和一些念珠菌属物种对CMN耐药。进一步的研究表明，CMN降低了毒力因子，如尿素酶活性，黑色素的产生，和生物膜的形成在Cne。CMN还抑制巨噬细胞内Cne感染。在靶特异性裂解纳米荧光素酶测定中，CMN的IC 50为4.6 μg/mL。CMN与重组Prp 8内含肽的结合通过热位移测定和微量热泳证明。用CMN处理Cne细胞减少内含肽剪接。CMN具有抑真菌作用，并与已知的抗真菌药物黄曲霉素B具有协同作用。最后，在隐球菌肺部感染小鼠模型中，20 mg/kg体重的CMN治疗导致肺部真菌负荷减少60%。总的来说，CMN代表了一种有效的抗真菌剂，具有治疗Cne和可能的Cga感染的新作用机制。
Drug resistance is a significant concern in the treatment of diseases, including cryptococcosis caused by Cryptococcus neoformans (Cne) and Cryptococcus gattii (Cga). Alternative drug targets are necessary to overcome drug resistance before it attains a critical stage. Splicing of inteins from proprotein precursors is crucial for activities of essential proteins hosting intein elements in many organisms, including human pathogens such as Cne and Cga. Through a high-throughput screening, we identified calcimycin (CMN) as a potent Prp8 intein splicing inhibitor with a minimum inhibitory concentration (MIC) of 1.5 μg/mL against the wild-type Cne-H99 (Cne-WT or Cne). In contrast, CMN inhibited the intein-less mutant strain (Cne-Mut) with a 16-fold higher MIC. Interestingly, Aspergillus fumigatus and a few Candida species were resistant to CMN. Further studies indicated that CMN reduced virulence factors such as urease activity, melanin production, and biofilm formation in Cne. CMN also inhibited Cne intracellular infection in macrophages. In a target-specific split nanoluciferase assay, the IC50 of CMN was 4.6 μg/mL. Binding of CMN to recombinant Prp8 intein was demonstrated by thermal shift assay and microscale thermophoresis. Treating Cne cells with CMN reduced intein splicing. CMN was fungistatic and showed a synergistic effect with the known antifungal drug amphotericin B. Finally, CMN treatment at 20 mg/kg body weight led to 60% reduction in lung fungal load in a cryptococcal pulmonary infection mouse model. Overall, CMN represents a potent antifungal with a novel mechanism of action to treat Cne and possibly Cga infections.
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影响因子：
16.6
作者：
Billmyre, R. Blake;Clancey, Shelly Applen;Heitman, Joseph
通讯作者：
Heitman, Joseph
DOI：
10.4049/jimmunol.1700958
发表时间：
2018-07-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
De Leon-Rodriguez CM;Rossi DCP;Fu MS;Dragotakes Q;Coelho C;Guerrero Ros I;Caballero B;Nolan SJ;Casadevall A
通讯作者：
Casadevall A
影响因子：
2.9
作者：
Capilla, Javier;Maffei, Claudia M. L.;Stevens, David A.
通讯作者：
Stevens, David A.
影响因子：
3.7
作者：
Arras SD;Chitty JL;Blake KL;Schulz BL;Fraser JA
通讯作者：
Fraser JA
影响因子：
2.9
作者：
Cordeiro, Rossana A.;Teixeira, Carlos E. C.;Rocha, Marcos F. G.
通讯作者：
Rocha, Marcos F. G.