Trichophyton rubrum transcriptome in response to acriflavine. Trichophyton rubrum

Purpose: The dermatophyte Trichophyton rubrum is an anthropophilic filamentous fungus that infects keratinized tissues and is the most common etiologic agent isolated in cases of human dermatophytoses. To better understand the molecular effects of stress responses and fungal adaptability, we evaluated the effects of acriflavine, a cytoxic drug, on T. rubrum transcriptome in a time-course assay using high-throughput RNA-seq technology. Results: RNA-seq generated approximately 200 million short reads that were mapped to the Broad Institute’s Dermatophyte Comparative Database before differential gene expression analysis. A subset of 490 genes modulated in response to stress caused T. rubrum exposure to acriflavine were identified. These genes are involved in various cellular processes such as oxidation-reduction reactions, transmembrane transport, metal ion binding, and pathogenicity. The genes involved in pathogenicity were down-regulated, suggesting that this drug interferes with virulence factors that allow the establishment and maintenance of host infection. Conclusion: The results obtained in this large-scale analysis provide insights into the molecular events underlying the stress responses of T. rubrum Acriflavine. Overall design: mRNA profiling of T. rubrum grown in Sabouraud media as control (0h) and three different times of exposure to sub-inhibitory acriflavine concentrations for 3h, 12h, and 24h were used for high-throughput RNA sequencing using Applied Biosystems Solid 4 System.

研究目的：红色毛癣菌（Trichophyton rubrum）是一类嗜人源性丝状真菌，可侵染人体角化组织，是人类皮肤癣菌病分离样本中最常见的致病原。为深入解析应激反应与真菌适应性的分子效应，本研究采用高通量RNA测序（RNA-seq）技术开展时间梯度实验，评估细胞毒性药物吖啶黄素（acriflavine）对红色毛癣菌转录组的影响。
研究结果：RNA-seq共产生约2亿条短读长序列，在开展差异基因表达分析前，将这些序列比对至布罗德研究所（Broad Institute）皮肤癣菌比较数据库。最终鉴定出490个在红色毛癣菌暴露于吖啶黄素所引发的应激反应中受到调控的基因。这些基因参与多种细胞生物学过程，包括氧化还原反应、跨膜运输、金属离子结合以及致病性相关通路。其中，致病性相关基因呈下调表达，提示该药物可干扰宿主感染建立与维持所需的毒力因子。
研究结论：本大规模分析所得结果为红色毛癣菌暴露于吖啶黄素后的应激反应相关分子事件提供了新的研究视角。
整体实验设计：以沙氏培养基（Sabouraud media）中培养的红色毛癣菌为对照组（0小时），设置3小时、12小时、24小时三个时间梯度，将红色毛癣菌暴露于亚抑制浓度的吖啶黄素环境中，随后采用应用生物系统Solid 4测序平台（Applied Biosystems Solid 4 System）开展高通量RNA测序，进行mRNA表达谱分析。