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 5

ALDH inhibition decreases collagen production, increases matrix contraction and proliferation, and normalizes αSMA organization of OMMP fibroblasts in vitro.

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ALDH inhibition decreases collagen production, increases matrix contract...
(A) Concentration of collagen from F-C (n = 4), F-PemI (n = 4), and F-PemU (n = 4) treated with ALDH inhibitors DEAB, disulfiram, or vehicle. Collagen type 1 was also measured in F-C fibroblasts treated with the ALDH metabolite, retinoic acid, or vehicle. (B) Western blotting for (top panel) COL1 and COL3 basal- and disulfiram-treated levels for F-PemI with densitometry panels for both F-PemI and F-PemU (see Supplementary Figure 4 for F-PemU gels). Lower panel shows COL1 basal levels for F-C (n = 3) and ATRA treated F-C (n = 3). (C) Area contracted at 72 hours by F-C (n = 4), F-PemI (n = 4), or F-PemU (n = 4) fibroblasts within a free-floating collagen matrix treated with DEAB or disulfiram compared with vehicle, also shown for F-C fibroblasts treated with retinoic acid. (D) Fluorescence readings (counts per second [cps]) from F-C (n = 4), F-PemI (n = 4), or F-PemU (n = 4) treated with DEAB, disulfiram, or vehicle and from F-C fibroblasts treated with retinoic acid. (E) Representative images of αSMA staining in F-PemI (n = 4) or F-PemU (n = 4) fibroblasts treated with either DEAB (not shown) or disulfiram within tethered collagen matrices. The top panel shows ALDH inhibition increased αSMA organization (compare with normal F-C controls in Figure 4E). Retinoic acid treatment of F-C (n = 4) fibroblasts in a tethered collagen matrix resulted in decreased αSMA organization (bottom panel), similar to that seen in OMMP fibroblasts (as shown in Figure 4E with F-PemI or F-PemU). Each assay was carried out with 4 separate patient fibroblast cultures and at least 3 technical repeats. Error bars represent ±SEM. *P < 0.05, **P < 0.005, ****P < 0.00005 as calculated using one-way ANOVA with Bonferroni correction.