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

Abstract

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.

• Authors: Mayumi Ueta, Hiromi Nishigaki, Chie Sotozono, Norihiko Yokoi, Katsura Mizushima, Yuji Naito, Shigeru Kinoshita
• Journal: Scientific reports (Sci Rep) Vol. 10 Issue 1 Pg. 17239 (10 14 2020) ISSN: 2045-2322 [Electronic] England
• PMID: 33057072 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• ANKRD1 protein, human
• CXCL2 protein, human
• CXCL8 protein, human
• Chemokine CXCL2
• Interleukin-8
• MIRN455 microRNA, human
• MicroRNAs
• Muscle Proteins
• Nuclear Proteins
• Repressor Proteins

Topics

• Adult
• Chemokine CXCL2 (genetics, metabolism)
• Conjunctiva (metabolism)
• Epithelium (metabolism)
• Humans
• Interleukin-8 (genetics, metabolism)
• MicroRNAs (genetics, metabolism)
• Middle Aged
• Muscle Proteins (genetics, metabolism)
• Nuclear Proteins (genetics, metabolism)
• Repressor Proteins (genetics, metabolism)
• Stevens-Johnson Syndrome (genetics, metabolism)