Dual RNA-Seq Analysis of Trichophyton rubrum and HaCat Keratinocyte Co-Culture Highlights Important Genes for Fungal-Host Interaction

Purpose: Evaluate the transcriptional profile of genes involved in fungus-host interactions using RNA sequencingMethods: T. rubrum and HaCat cells line were co-cultured in RPMI medium supplemented with 5% of fetal bovine serum and were incubated for 24 h at 37ºC in a humidified atmosphere containing 5% CO2 , followed RNA by extraction of both organisms, libraries construction and sequence.Results: Our data demonstrated the induction of specific genes that may improve the assimilation of nutrients and fungal survival in the host and genes encoding keratinolytic proteases that are important for T. rubrum virulence during infection. In HaCat cell lines it were induced genes encoding antimicrobial activity , cell migration and epithelial barrier repair. Futhermore, the genes KRT1 and FLG involved in epithelial barrier integrity were repressedConclusions: This mixed transcriptome analysis showed the modulation of important genes involved in the mechanism of T. rubrum infection and in the defense and maintenance of cell homeostasis of keratinocytes, which could represent potential antifungal targets for new therapeutic approaches to the treatment of dermatophytoses

研究目的：利用RNA测序（RNA sequencing）评估参与真菌-宿主互作的基因的转录谱。
研究方法：将红色毛癣菌（T. rubrum）与HaCat细胞系在添加了5%胎牛血清的RPMI培养基中进行共培养，于37℃、含5%二氧化碳的湿润培养箱中孵育24小时；随后提取两种生物的总RNA，构建测序文库并进行测序。
研究结果：本研究数据显示，部分特异性基因的表达被上调，这类基因可增强真菌在宿主内的营养同化能力与存活能力；另有部分编码角质溶解蛋白酶的基因，其表达上调对红色毛癣菌感染过程中的毒力发挥具有关键作用。在HaCat细胞系中，编码抗菌活性相关蛋白、参与细胞迁移与上皮屏障修复的基因表达被上调。此外，参与上皮屏障完整性维持的KRT1与FLG基因的表达被下调。
研究结论：本混合转录组分析揭示了参与红色毛癣菌感染机制的重要基因，以及角质形成细胞防御与细胞稳态维持相关基因的表达调控模式，该发现可为开发治疗皮肤癣菌病的新型抗真菌治疗策略提供潜在靶点。