Responses of keratinocytes to Trichophyton mentagrophyte infection based on whole transcriptome analysis.

IF 4.1 2区医学 Q1 DERMATOLOGY

Mycoses Pub Date : 2024-03-01 DOI:10.1111/myc.13713

Fangfang Zang, Zhangxu Wang, Yuanxue Yang, Ziyao Xiao, Zhaodan Wu, Jia Zhang, Bishibei Li, Jiayu Zheng, Xixi Zhu, Ying Wan, Ran Wang, Wudian Xiao

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引用次数: 0

Abstract

Background: Dermatophytosis is an intractable superficial mycosis in humans and animals mainly caused by Trichophyton mentagrophytes (T. mentagrophytes), with a global prevalence of about 20%. Keratinocytes are the most abundant participants in skin immunity, and they also play a role in the first-line defence against T. mentagrophytes. However, no studies of keratinocyte responses against T. mentagrophytes infection based on the whole transcriptome have been reported.

Objectives: Here, we systematically analysed changes in keratinocytes infected with T. mentagrophytes using whole transcriptome sequencing technology.

Methods: The phenotypic changes in keratinocytes after infection with 1 × 10⁵ conidia/mL T. mentagrophytes were observed by light microscopy, scanning electron microscopy, transmission electron microscopy and terminal deoxynucleotidyl transferase dUTP nick end labeling. RNA-sequencing (RNA-seq), small RNA-seq technology and related bioinformatics methods were used to systematically analyse the whole transcriptome changes in keratinocytes upon T. mentagrophytes stimulation.

Results: We found that T. mentagrophytes infection caused morphological changes, membrane damage, the formation of irregular organelles and keratinocyte apoptosis. A total of 204 differentially expressed (DE) circular RNAs (circRNAs), 868 DE long noncoding RNAs (lncRNAs), 2973 DE mRNAs and 209 DE micro RNAs (miRNAs) were identified between noninfected and T. mentagrophytes-infected keratinocytes. The expression level of selected RNAs was validated by quantitative real-time polymerase chain reaction (qRT-PCR). Functional enrichment analysis revealed that the parental genes of DE circRNAs were related to cell response, cell death and establishment of the skin barrier. Genes targeted by miRNA were involved in regulating the initiation of the immune response. Based on the expression level of circRNAs, lncRNAs, mRNAs and miRNAs, circRNA-miRNA-mRNA competing endogenous (ceRNA) networks comprised of 159 DE miRNAs, 141 DE circRNAs and 2307 DE mRNAs, and lncRNA-miRNA-mRNA ceRNA networks comprised of 790 DE lncRNAs, 190 DE miRNAs and 2663 DE mRNAs were constructed. The reliability of two selected ceRNA networks was verified using qRT-PCR. Further functional enrichment analysis revealed that the DE mRNAs interacting with circRNAs and lncRNAs in the ceRNA network mainly participated in fungal recognition, inflammation, the innate immune response and the death of keratinocytes.

Conclusions: Our findings might provide new evidence on the pathogenesis of T. mentagrophytes-induced dermatophytosis, which is essential for identifying new therapeutic targets for dermatophytosis treatment.

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基于全转录组分析的角质细胞对毛癣菌感染的反应

背景：皮癣是人类和动物身上的一种难治性浅表真菌病，主要由曼陀罗菌（T. mentagrophytes）引起，全球发病率约为 20%。角质形成细胞是皮肤免疫系统中最丰富的参与者，它们也是抵御脑毛囊线虫的第一道防线。然而，目前还没有基于整个转录组研究角质细胞对传染性单胞菌感染的反应的报道：在此，我们利用全转录组测序技术系统分析了感染了脑膜炎双球菌的角朊细胞的变化：方法：用光学显微镜、扫描电子显微镜、透射电子显微镜和末端脱氧核苷酸转移酶 dUTP 缺口标记法观察感染 1 × 105 分生孢子/毫升 T. mentagrophytes 后角质细胞的表型变化。利用RNA测序（RNA-seq）、小RNA-seq技术和相关生物信息学方法，系统分析了在T. mentagrophytes刺激下角质形成细胞整个转录组的变化：结果：我们发现，T. mentagrophytes 感染会导致角朊细胞形态改变、膜损伤、不规则细胞器形成和角朊细胞凋亡。在未感染和感染了曼陀罗菌的角朊细胞之间共发现了204个差异表达（DE）的环状RNA（circRNAs）、868个差异表达（DE）的长非编码RNA（lncRNAs）、2973个差异表达（DE）的mRNAs和209个差异表达（DE）的微RNAs（miRNAs）。通过实时定量聚合酶链反应（qRT-PCR）验证了所选 RNA 的表达水平。功能富集分析表明，DE circRNA 的亲代基因与细胞反应、细胞死亡和皮肤屏障的建立有关。miRNA靶向的基因参与调节免疫反应的启动。根据circRNAs、lncRNAs、mRNAs和miRNAs的表达水平，构建了由159个DE miRNAs、141个DE circRNAs和2307个DE mRNAs组成的circRNA-miRNA-mRNA竞争内源（ceRNA）网络，以及由790个DE lncRNAs、190个DE miRNAs和2663个DE mRNAs组成的lncRNA-miRNA-mRNA ceRNA网络。利用 qRT-PCR 验证了两个选定 ceRNA 网络的可靠性。进一步的功能富集分析表明，ceRNA网络中与circRNAs和lncRNAs相互作用的DE mRNAs主要参与真菌识别、炎症、先天性免疫反应和角质形成细胞的死亡：我们的研究结果可能为曼地夫球菌诱导的皮癣病的发病机制提供了新的证据，这对于确定皮癣病治疗的新靶点至关重要。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Mycoses 医学-皮肤病学

CiteScore

10.00

自引率

8.20%

发文量

143

审稿时长

6-12 weeks

期刊介绍： The journal Mycoses provides an international forum for original papers in English on the pathogenesis, diagnosis, therapy, prophylaxis, and epidemiology of fungal infectious diseases in humans as well as on the biology of pathogenic fungi. Medical mycology as part of medical microbiology is advancing rapidly. Effective therapeutic strategies are already available in chemotherapy and are being further developed. Their application requires reliable laboratory diagnostic techniques, which, in turn, result from mycological basic research. Opportunistic mycoses vary greatly in their clinical and pathological symptoms, because the underlying disease of a patient at risk decisively determines their symptomatology and progress. The journal Mycoses is therefore of interest to scientists in fundamental mycological research, mycological laboratory diagnosticians and clinicians interested in fungal infections.