Regulation of gene expression by miRNA-455-3p, upregulated in the conjunctival epithelium of patients with Stevens–Johnson syndrome in the chronic stage

• Published in: Scientific reports Vol. 10; no. 1; p. 17239
• Main Authors: Ueta, Mayumi, Nishigaki, Hiromi, Sotozono, Chie, Yokoi, Norihiko, Mizushima, Katsura, Naito, Yuji, Kinoshita, Shigeru
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 14.10.2020
• Subjects: 692/308/53; 692/699/3161/3162; 692/699/3161/3163; Adult; Chemokine CXCL2 - genetics; Chemokine CXCL2 - metabolism; Conjunctiva - metabolism; Epithelium - metabolism; Humanities and Social Sciences; Humans; Interleukin-8 - genetics; Interleukin-8 - metabolism; MicroRNAs - genetics; MicroRNAs - metabolism; Middle Aged; multidisciplinary; Muscle Proteins - genetics; Muscle Proteins - metabolism; Nuclear Proteins - genetics; Nuclear Proteins - metabolism; Repressor Proteins - genetics; Repressor Proteins - metabolism; Science; Science (multidisciplinary); Stevens-Johnson Syndrome - genetics; Stevens-Johnson Syndrome - metabolism
• DOI: 10.1038/s41598-020-74211-9
• ISSN: 2045-2322

To investigate the role of miRNA in the pathogenesis underlying ocular surface complications in patients with Stevens–Johnson syndrome (SJS)/toxic epidermal necrolysis (TEN) in the chronic stage. Using oligonucleotide microarrays, we performed comprehensive miRNA analysis of the conjunctival epithelium of SJS/TEN patients with severe ocular complications (SOC) in the chronic stage (n = 3). Conjunctival epithelium of patients with conjunctival chalasis (n = 3) served as the control. We confirmed the down- and up-regulation of miRNA of interest by quantitative real-time polymerase chain reaction (RT-PCR) assays using the conjunctival epithelium from 6 SJS/TEN with SOC patients and 7 controls. We focused on miRNA-455-3p, which is significantly upregulated in the conjunctival epithelium of the SJS/TEN patients, and investigated its function by inhibiting miR-455-3p in primary human conjunctival epithelial cells (PHCjEs). Comprehensive miRNA expression analysis showed that the expression of 5 kinds of miRNA was up-regulated more than fivefold, and that the expression of another 5 kinds of miRNA was down-regulated by less than one-fifth. There was a significant difference between the SJS/TEN patients and the controls [analysis of variance (ANOVA) p < 0.05]. Quantitative miRNA PCR assay showed that hsa-miR-31* and hsa-miR-455-3p were significantly up-regulated in the conjunctival epithelium of the SJS/TEN patients. Comprehensive gene expression analysis of PHCjEs transfected with the hsa-miR-455-3p inhibitor and quantitative RT PCR assay showed that ANKRD1, CXCL8, CXCL2, GEM, PTGS2, RNASE8, IL6, and CXCL1 were down-regulated by the hsa-miR-455-3p inhibitor. Quantitative RT-PCR, focused on the genes that tended to be up-regulated in SJS/TEN with SOC, revealed that the expression of IL1A, KPRP, IL36G, PPP1R3C, and ADM was significantly down-regulated in PHCjEs transfected with the hsa-miR-455-3p inhibitor. Our results suggest that miRNA-455-3p could regulate many genes including innate immune related genes in human conjunctival epithelium, and that its up-regulation contributes to the pathogenesis on the ocular surface in SJS/TEN patients with the SOC in the chronic stage. Our findings may lead to the development of new treatments using the miRNA-455-3p inhibitor.