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Sarah B. Dale, Valerie P. Saw, John K. Dart, David J. Abraham, Julie T. Daniels; Differential Expression Of Genes In Conjunctival Tissue, And Primary Conjunctival Fibroblasts, From Mucous Membrane Pemphigoid Patients. Invest. Ophthalmol. Vis. Sci. 2012;53(14):2704.
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© ARVO (1962-2015); The Authors (2016-present)
Conjunctival gene expression profiles of patients with ocular mucous membrane pemphigoid (MMP) with (PemI) or without (PemU) inflammation and controls (C) were compared to identify any differential expression. These gene expression profiles were also compared to those of primary conjunctival fibroblasts explanted from PemI (F-PemI), PemU (F-PemU), and C (F-C) conjunctival tissue which are the focus of our in vitro model of fibroblast activity in MMP.
Total RNA from conjunctival tissue of 12 MMP patients and 5 age-matched controls (PemI, n=6; PemU, n=6; C, n=5) and from primary conjunctival fibroblasts from 4 MMP patients and 2 controls (F-PemI, n=2; F-PemU, n=2; F-C, n=2) was isolated with RNeasy Micro Kit. The MMP patients all had biopsy confirmed MMP (PemI, n=12; PemU, n=4). Quality and concentration of the total RNA was analysed using the NanoDrop ND-1000 and a Bioanalyser. Microarray experiments were performed at Cambridge Genomic Services, using the HumanHT-12 v4 BeadChip.
65 genes are showing at least a 2 fold increased expression within PemI and PemU tissue in comparison with C tissue. A selection of these genes are involved with inflammatory, wound healing and immune responses, and their increased expression differs between PemI and PemU tissue. These genes are also expressed in the F-PemI, F-PemU and F-C primary fibroblasts.
MMP conjunctival tissue exhibits a differentially expressed gene profile compared to healthy conjunctiva. Elements of this profile are maintained in-vitro within MMP primary fibroblasts. These findings support the utility of our in-vitro model of fibroblast activity in MMP, through which scarring mechanisms in MMP may be elucidated, and anti-scarring strategies investigated.
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