Expression Profile Analysis Of MRNAs,lncRNAs And MiRNAs In Candida Albicans ?-glucan-stimulated Monocytes/macrophage And The Study Of The Regulatory Mechanisms Of MiR-210

Background:Candida albicans(C.albicans)is an opportunistic human pathogen that can colonize the surfaces of the skin and mucosa in healthy individuals.In healthy individuals,C.albicans is usually harmless,and the immune system can effectively resist its invasion.However,C.albicans becomes pathogenic in immunocompromised patients.With the increase in the immune-suppressed population,C.albicans contributes to more than half of cases of superficial and systemic candidiasis.The eukaryotic nature of C.albicans and the challenges of drug resistance,emphasize the difficulties of treatment.To explore more therapeutic targets for the treatment of candidiasis,the pathological mechanism of C.albicans infection should be investigated in depth.Innate immunity serves as the first line of defense against C.albicans.Innate immune cells include monocytes,macrophages,dendritic cells,neutrophil,etc.The C.albicans cell wall consists of approximately 60%?-glucan.The dendritic cell-associated C-type lectin-1(dectin-1)can mediate innate immune response through spleen tyrosine kinase and caspase recruitment domain protein 9(Syk-CARD9)-dependent manner,stimulation of NF-?B and p38 MAPK signaling pathway.Noncoding RNA include long noncoding RNAs(lncRNAs)and microRNAs(miRNAs),etc.Emerging evidence suggests that lncRNAs play important roles in disease development.However,the roles of lncRNAs in the pathogenesis of C.albicans remain unclear.Our previous study identify three differentially expressed(DE)miRNAs in THP-1 cells after dectin-1 triggered by insoluble ?-glucan from the cell wall of C.albicans(CaIG),including miR-210,miR-146a and miR-30.The roles of miR-210 in the innate immune of C.albicans infection remain unclear.Objectives:1.Our study aimed to investigate and characterize the mRNA and lncRNA transcriptomes of CD14+ monocytes and THP-1 cells stimulated with CaIG by RNA-seq.2.To study the regulation mechanism of miR-210 in dectin-1-ligand(CaIG)-induced inflammation in THP-1 cells.Methods:1.We use RNA-seq to detected DE mRNAs and lncRNAs between CD14+ monocytes which were isolated from healthy donors' blood and THP-1 cells with or without stimulated by CaIG.2.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were performed to describe the function of DE genes and to identify potential signaling pathways involved in the pathogenesis of C.albicans infection.3.A possible IncRNA-mRNA coexpression network were performed to predict a preliminary function of DE lncRNAs.4.We use bioinformatics methods to predict the target genes of DE miRNAs between THP-1 cells with or without stimulated by CaIG,and GO analyses were performed for the target genes.5.We transfect THP-1 cells with miR-210 mimic or inhibitor to study the regulation mechanism of miR-210 in dectin-1-ligand(CaIG)-induced inflammatory response.Results:1.A total of 10788 DE mRNAs and 2021 DE lncRNAs were obtained from CaIG-treated CD14+monocytes compared with untreated CD14+monocytes.In the comparison between CaIG-treated and untreated THP-1 cells,we found 3349 DE mRNAs and 291 DE lncRNAs.Intersection analysis of mRNA and lncRNA profiling between the two groups showed that 808 dysregulated mRNAs and 51 dysregulated lncRNAs overlapped between CaIG-treated CD14+monocytes and THP-1 cells.We examined five collectively DE mRNAs and lncRNAs in both cells using quantitative real-time PCR,validating the reliability of the RNA-seq results.2.GO and KEGG pathway analyses revealed that the 808 common DE mRNAs were mostly enriched in the inflammatory response and NF-?B signaling pathway,respectively.3.LncRNA-mRNA coexpression analysis was performed for the 51 DE lncRNAs and the 808 DE mRNAs in the two groups(the absolute value of PCC>0.99,p<0.001).We chose the common network pairs of the two groups to construct the coexpression network and revealed 97 network pairs comprising 8 dysregulated lncRNAs and 60 dysregulated mRNAs.We speculated that lncRNA 1nc-CCL3L3-1:1 might be involved in the NF-?B signaling pathway in CaIG induced innate immune response of monocytes/macrophages.4.GO analyses revealed that the target genes of miRNA-146a,miRNA-210 and miRNA-30 miRNAs were mostly enriched in nucleus.5.We found CaIG-induced the expression of miR-210 in THP-1 cells in a time-dependent manner.MiR-210 was strongly upregulated and the response peaked at 24 hours.MiR-210 markedly downregulate CaIG-induced expression and production of TNF-? and IL-6 in THP-1 cells.MiR-210 inhibit CaIG-induced activity of Syk,I?B? and translocation of NF-?B p65 in THP-1 cells.Conclusions:1.Our results emphasized the potential roles of lncRNAs in the innate immunity of C.albicans infection.2.Dectin-1-ligand(CaIG)-induced miR-210 acts as a negative feedback regulator of inflammation through down-regulation of the Syk-NF-?B pathway in THP-1 cells following dectin-1 stimulation.