Pitangui Nayla de Souza, de Lacorte Singulani Junya, Sardi Janaina de Cássia Orlandi, de Souza Paula Carolina, Rodríguez-Arellanes Gabriela, García-Pérez Blanca Estela, Enguita Francisco Javier, Pavan Fernando R, Taylor Maria Lucia, Mendes-Giannini Maria José Soares, Fusco-Almeida Ana Marisa
School of Pharmaceutical Sciences, São Paulo State University (UNESP), Araraquara, São Paulo 14800-903, Brazil.
Department of Cellular and Molecular Biology, Ribeirão Preto School of Medicine, University of São Paulo, Ribeirão Preto, São Paulo 14049-900, Brazil.
J Fungi (Basel). 2021 Jan 18;7(1):60. doi: 10.3390/jof7010060.
affects healthy and immunocompromised individuals, sometimes causing a severe disease. This fungus has two morphotypes, the mycelial (infective) and the yeast (parasitic) phases. MicroRNAs (miRNAs) are small RNAs involved in the regulation of several cellular processes, and their differential expression has been associated with many disease states. To investigate miRNA expression in host cells during infection, we studied the changes in the miRNA profiles of differentiated human macrophages infected with yeasts from two fungal strains with different virulence, EH-315 (high virulence) and 60I (low virulence) grown in planktonic cultures, and EH-315 grown in biofilm form. MiRNA profiles were evaluated by means of reverse transcription-quantitative polymerase chain reaction using a commercial human miRNome panel. The target genes of the differentially expressed miRNAs and their corresponding signaling pathways were predicted using bioinformatics analyses. Here, we confirmed biofilm structures were present in the EH-315 culture whose conditions facilitated producing insoluble exopolysaccharide and intracellular polysaccharides. In infected macrophages, bioinformatics analyses revealed especially increased (hsa-miR-99b-3p) or decreased (hsa-miR-342-3p) miRNAs expression levels in response to infection with biofilms or both growth forms of yeasts, respectively. The results of miRNAs suggested that infection by can affect important biological pathways of the host cell, targeting two genes: one encoding a protein that is important in the cortical cytoskeleton; the other, a protein involved in the formation of stress granules. Expressed miRNAs in the host's response could be proposed as new therapeutic and/or diagnostic tools for histoplasmosis.
可影响健康个体和免疫功能低下个体,有时会引发严重疾病。这种真菌有两种形态型,即菌丝体(感染性)和酵母(寄生性)阶段。微小RNA(miRNA)是参与多种细胞过程调控的小RNA,其差异表达与许多疾病状态相关。为了研究感染期间宿主细胞中miRNA的表达情况,我们研究了用两种毒力不同的真菌菌株(浮游培养的EH - 315(高毒力)和60I(低毒力)以及生物膜形式生长的EH - 315)的酵母感染分化的人巨噬细胞后miRNA谱的变化。使用商业人类miRNome检测板通过逆转录定量聚合酶链反应评估miRNA谱。使用生物信息学分析预测差异表达的miRNA的靶基因及其相应的信号通路。在此,我们证实了EH - 315培养物中存在生物膜结构,其条件有利于产生不溶性胞外多糖和细胞内多糖。在感染的巨噬细胞中,生物信息学分析显示,分别响应生物膜感染或两种酵母生长形式的感染,miRNA表达水平特别增加(hsa - miR - 99b - 3p)或降低(hsa - miR - 342 - 3p)。miRNA的结果表明,[真菌名称未给出]感染可影响宿主细胞的重要生物学途径,靶向两个基因:一个编码在皮质细胞骨架中起重要作用的蛋白质;另一个是参与应激颗粒形成的蛋白质。宿主反应中表达的miRNA可被提议作为组织胞浆菌病的新治疗和/或诊断工具。