Aldehyde dehydrogenase inhibition blocks mucosal fibrosis in human and mouse ocular scarring
Sarah D. Ahadome, … , Julie T. Daniels, John K. Dart
Sarah D. Ahadome, … , Julie T. Daniels, John K. Dart
Published August 4, 2016
Citation Information: JCI Insight. 2016;1(12):e87001. https://doi.org/10.1172/jci.insight.87001.
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Research Article Inflammation Ophthalmology

Aldehyde dehydrogenase inhibition blocks mucosal fibrosis in human and mouse ocular scarring

Abstract

Mucous membrane pemphigoid (MMP) is a systemic mucosal scarring disease, commonly causing blindness, for which there is no antifibrotic therapy. Aldehyde dehydrogenase family 1 (ALDH1) is upregulated in both ocular MMP (OMMP) conjunctiva and cultured fibroblasts. Application of the ALDH metabolite, retinoic acid (RA), to normal human conjunctival fibroblasts in vitro induced a diseased phenotype. Conversely, application of ALDH inhibitors, including disulfiram, to OMMP fibroblasts in vitro restored their functionality to that of normal controls. ALDH1 is also upregulated in the mucosa of the mouse model of scarring allergic eye disease (AED), used here as a surrogate for OMMP, in which topical application of disulfiram decreased fibrosis in vivo. These data suggest that progressive scarring in OMMP results from ALDH/RA fibroblast autoregulation, that the ALDH1 subfamily has a central role in immune-mediated ocular mucosal scarring, and that ALDH inhibition with disulfiram is a potential and readily translatable antifibrotic therapy.

Authors

Sarah D. Ahadome, David J. Abraham, Suryanarayana Rayapureddi, Valerie P. Saw, Daniel R. Saban, Virginia L. Calder, Jill T. Norman, Markella Ponticos, Julie T. Daniels, John K. Dart

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Figure 4

Primary conjunctival fibroblasts from OMMP patients maintain a fibrotic phenotype in vitro.

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Primary conjunctival fibroblasts from OMMP patients maintain a fibrotic ...
(A) Concentration of collagen from ELISA and Sircol assays from F-C (n = 4), F-PemI (n = 4), and F-PemU (n = 4) fibroblasts. (B) Basal COL1 levels were also measured by Western blotting for F-C (n = 3), F-PemU (n = 3), and F-PemI (n = 3); the densitometry shows significant differences for F-PemI compared with F-C fibroblasts. There is a trend for a difference between F-PemU and F-C, but this was not statistically significant. (C) Area contracted at 72 hours by F-C (n = 4), F-PemI (n = 4), or F-PemU (n = 4) fibroblasts in free-floating collagen gels. The arrows indicate the edge of the gels. (D) Proliferation measured by fluorescence intensity (counts per second [cps]) from F-C (n = 4), F-PemI (n = 4), or F-PemU (n = 4) fibroblasts labeled with the fluorescent CyQuant nucleotide dye. (E) Tethered collagen gels populated with F-C (n = 4), F-PemI (n = 4), or F-PemU (n = 4) fibroblasts were stained for αSMA (red) and nuclei stained with DAPI (blue). Each assay was carried out with 4 separate patient fibroblast cultures and at least 3 technical repeats. (F) αSMA levels were also measured by Western blotting and densitometry for basal F-C (n = 3), F-PemU (n = 3), and F-PemI (n = 3). Error bars represent mean ±SEM. *P < 0.05; **P < 0.005; ****P < 0.00005 as calculated using one-way ANOVA with Bonferroni correction. Scale bar: 100 μm.