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.

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Gene expression analysis of conjunctival epithelium of patients with Stevens-Johnson syndrome in the chronic stage

BMJ Open Ophthalmology ◽

10.1136/bmjophth-2018-000254 ◽

2019 ◽

Vol 4 (1) ◽

pp. e000254 ◽

Cited By ~ 1

Author(s):

Mayumi Ueta ◽

Chie Sotozono ◽

Hiromi Nishigaki ◽

Suzuko Ohsako ◽

Norihiko Yokoi ◽

...

Keyword(s):

Gene Expression ◽

Expression Analysis ◽

Ocular Surface ◽

Gene Expression Analysis ◽

Surface Protein ◽

Stevens Johnson Syndrome ◽

Conjunctival Epithelium ◽

Rt Pcr ◽

Chronic Stage ◽

Johnson Syndrome

ObjectiveTo investigate the pathology underlying the ocular surface complications of patients with Stevens-Johnson syndrome (SJS) in the chronic stage.Methods and analysisUsing oligonucleotide microarrays, we performed comprehensive gene expression analysis of the conjunctival epithelium of patients with SJS in the chronic stage (n=3). The controls were patients with conjunctival chalasis (n=3). We confirmed the downregulation and upregulation of transcripts of interest by quantitative real-time PCR (RT-PCR) assay. The expression of ocular surface protein with significantly upregulated transcripts was assessed immunohistochemically.ResultsCompared with the controls, in the conjunctival epithelium of patients with SJS, 50 transcripts were downregulated by less than one-tenth (analysis of variance (ANOVA) p<0.05). Transcripts MUC7, PIGR, HEPACAM2, ADH1C and SMR3A were downregulated by less than one-fiftieth. 65 transcripts were upregulated more than 10- fold; the difference between patients with SJS and the controls was significant (ANOVA p<0.05). There were 14 transcripts that were upregulated more than 50-fold; they were SERPINB4, KRT1, KRTDAP, S100A7, SBSN, KLK6, SERPINB12, PNLIPRP3, CASP14, ODZ2, CA2, CRCT1, CWH43 and FLG. Quantitative RT-PCR of conjunctival epithelium samples from 11 patients with SJS and 26 controls showed that the gene expression of PIGR, HEPACAM2 and ADH1C was significantly downregulated while the gene expression of ODZ2 (teneurin-2) was significantly upregulated in patients with SJS. We document that teneurin-2 protein can be expressed in human conjunctival epithelium.ConclusionOur results suggest that the downregulation of PIGR, HEPACAM2 and ADH1C and upregulation of teneurin-2 expression contribute to the pathology of the ocular surface in patients with SJS in the chronic stage.

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Pathogenesis of Stevens-Johnson Syndrome/Toxic Epidermal Necrolysis With Severe Ocular Complications

Frontiers in Medicine ◽

10.3389/fmed.2021.651247 ◽

2021 ◽

Vol 8 ◽

Author(s):

Mayumi Ueta

Keyword(s):

Toxic Epidermal Necrolysis ◽

Ocular Surface ◽

Acute Stage ◽

Stevens Johnson Syndrome ◽

Prostaglandin E ◽

Ocular Complications ◽

Chronic Stage ◽

Snp Association ◽

Johnson Syndrome ◽

Burn Units

Stevens-Johnson syndrome (SJS)/toxic epidermal necrolysis (TEN) is an acute inflammatory vesiculobullous reaction of the mucosa of the ocular surface, oral cavity, and genitals, and of the skin. Severe ocular complications (SOC) are observed in about half of SJS/TEN patients diagnosed by dermatologists and in burn units. Ophthalmologists treat SOC, and they tend to encounter the patients not only in the acute stage, but also in the chronic stage. Our investigation of the pathogenesis of SJS/TEN with SOC led us to suspect that abnormal innate mucosal immunity contributes to the ocular surface inflammation seen in SJS/TEN with SOC. We confirmed that cold medicines such as NSAIDs and multi-ingredient cold medications are the main causative drugs for SJS/TEN with SOC. Single nucleotide polymorphism (SNP) association analysis of cold medicine-related SJS/TEN with SOC showed that the Toll-like receptor 3 (TLR3)-, the prostaglandin-E receptor 3 (PTGER3)-, and the IKZF1 gene were significantly associated with SNPs and that these genes could regulate mucocutaneous inflammation including that of the ocular surface. We also examined the tear cytokines of SJS/TEN with SOC in the chronic stage and found that IL-8, IL-6, IFN-γ, RANTES, eotaxin, and MIP-1β were significantly upregulated in SJS/TEN with SOC in the chronic stage. Only IP-10 was significantly downregulated in SJS/TEN with SOC in the chronic stage. This mini-review summarizes the pathological mechanisms that we identified as underlying the development of SJS/TEN with SOC.

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Ocular surface cytokine profile in chronic Stevens-Johnson syndrome and its response to mucous membrane grafting for lid margin keratinisation

British Journal of Ophthalmology ◽

10.1136/bjophthalmol-2017-310373 ◽

2017 ◽

Vol 102 (2) ◽

pp. 169-176 ◽

Cited By ~ 12

Author(s):

Srividya Gurumurthy ◽

Geetha Iyer ◽

Bhaskar Srinivasan ◽

Shweta Agarwal ◽

Narayanasamy Angayarkanni

Keyword(s):

Gene Expression ◽

Growth Factor ◽

Mucous Membrane ◽

Ocular Surface ◽

Transforming Growth Factor ◽

Tissue Growth ◽

Stevens Johnson Syndrome ◽

Gm Csf ◽

Johnson Syndrome ◽

Collagen Iii

BackgroundTo study the tear cytokine and the conjunctival and oral mucosal marker profile in chronic ocular Stevens-Johnson syndrome (SJS) and their alteration following mucous membrane grafting (MMG) for lid margin keratinisation (LMK).MethodsIn a 1-year prospective study, SJS cases (n=25) and age-matched/sex-matched healthy controls (n=25) were recruited. Tear specimen (Schirmer’s strip), conjunctival and oral mucosal imprints were collected from controls and SJS cases pre-MMG and post-MMG (at first follow-up, n=17). Tear cytokines were profiled using 27-bioplex array. Transforming growth factor-beta (TGF-β)-mediated extracellular matrix changes in conjunctival and oral mucosal cells were analysed by gene expression studies. 30ResultsTear cytokine profiling of chronic SJS cases at pre-MMG stage revealed significant upregulation of cytokines granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-8, IL-1β, monocyte chemoattractant protein-1, IL-15, IL-2, IL-17A and basic fibroblast growth factor (bFGF) with downregulation of IP-10 (interferon gamma-induced protein 10), tumour necrosis factor-α, interferon-γ, IL-10, vascular endothelial growth factor, regulated upon activation normal T-cell expressed and secreted (RANTES), IL-7, IL-12p70 and IL-13, with maximal increase in GM-CSF and maximal downregulation of IP-10, respectively. Of these, IL-2, IL-15, bFGF and IL-17A showed significant correlation with disease severity, pre-MMG. Conjunctival cells pre-MMG showed increase in TGF-β1, TGF-βRII, connective tissue growth factor and collagen-III gene expression by 10, 67, 173 and 184 folds, respectively, which dropped to 1.3, 11, 13.5 and 19 folds correspondingly, post-MMG. However, their expressions in oral mucosa were negligible.ConclusionA proinflammatory, profibrotic, antiapoptotic ocular surface milieu characterises chronic ocular SJS. IP-10, an antifibrotic cytokine was noted to be maximally downregulated, unlike in other forms of chronic dry eye disease. The alterations in the ocular surface are seen to reverse largely with MMG for LMK.

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Comprehensive miRNA analysis of conjunctival epithelium of Stevens-Johnson syndrome patients in the chronic stage

World Allergy Organization Journal ◽

10.1016/j.waojou.2020.100257 ◽

2020 ◽

Vol 13 (8) ◽

pp. 100257

Author(s):

MAYUMI Ueta ◽

Hiromi Nishigaki ◽

Chie Sotozono ◽

Norihiko Yokoi ◽

Shigeru Kinoshita

Keyword(s):

Stevens Johnson Syndrome ◽

Conjunctival Epithelium ◽

Chronic Stage ◽

Johnson Syndrome

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Long-term Progression of Ocular Surface Disease in Stevens–Johnson Syndrome and Toxic Epidermal Necrolysis

Cornea ◽

10.1097/ico.0000000000002263 ◽

2020 ◽

Vol 39 (6) ◽

pp. 745-753

Author(s):

Yamato Yoshikawa ◽

Mayumi Ueta ◽

Hideki Fukuoka ◽

Tsutomu Inatomi ◽

Isao Yokota ◽

...

Keyword(s):

Toxic Epidermal Necrolysis ◽

Ocular Surface ◽

Ocular Surface Disease ◽

Stevens Johnson Syndrome ◽

Johnson Syndrome

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Altered retinoid metabolism gene expression in chronic Stevens-Johnson syndrome

British Journal of Ophthalmology ◽

10.1136/bjophthalmol-2018-312849 ◽

2019 ◽

Vol 103 (8) ◽

pp. 1015-1023 ◽

Cited By ~ 2

Author(s):

Gurumurthy Srividya ◽

Narayanasamy Angayarkanni ◽

Geetha Iyer ◽

Bhaskar Srinivasan ◽

Shweta Agarwal

Keyword(s):

Gene Expression ◽

Retinoic Acid ◽

Mucous Membrane ◽

Retinoic Acid Receptor ◽

Stevens Johnson Syndrome ◽

Bulbar Conjunctiva ◽

Ppar Γ ◽

Retinoid Metabolism ◽

Johnson Syndrome ◽

Alpha 1 Antitrypsin

BackgroundStevens-Johnson syndrome (SJS), a blistering disorder of the skin and mucous membrane, leads to ocular morbidity in >60% of cases. Retinoids are vital micronutrients for vision, regulating corneal and conjunctival cell proliferation, differentiation and immune function. This prospective case–control study probed for alterations in retinoid metabolism by evaluating retinoic acid receptor signalling in the conjunctival cells of patients with SJS.MethodsImprints were collected from the bulbar conjunctiva of patients with chronic SJS. The gene expression of retinoic acid receptors, namely, RXRA, RARA, RARG, RORA; the fibrosis marker TGFβ and its receptor TGFβRII; the transcription factors PPAR-γ, STRA6 and Stat3; the enzymes aldehyde dehydrogenase (ALDH1a1), alpha-1 antitrypsin (A1AT); and the Cyp genes Cyp26a1 and Cyp26b1 were assessed by quantitative PCR in patients with SJS pre-mucous (n = 34) and post-mucous membrane graft (MMG) intervention (n=19) in comparison with age-matched/sex-matched healthy controls (n=20). Western blot analysis of ALDH1a1, RARA and RARG were done in the conjunctival imprint cells.ResultsThe transcript levels of ALDH1a1, RXRA, RORA, STRA6, Cyp26a1 and Cyp26b1 were decreased around 4, 26, 17, 129, 9 and 8 folds, respectively, and RARA, RARG, PPAR-γ, TGFβ, TGFβRII were increased by 12, 15, 51, 16 and 87 folds, respectively, in SJS conjunctiva at the pre-MMG stage. The changes in RORA, Cyp26a1, Cyp26b1, RARA and Stat3 were statistically significant (p<0.05). Changes in protein expression of ALDH1a1, RARA and RARG supported the gene expression changes.ConclusionsThe study provides the first experimental insight into the role of retinoid metabolism in the ocular sequelae of chronic SJS.

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