Efficacy and safety of suberoylanilide hydroxamic acid (Vorinostat) in the treatment of canine corneal fibrosis.
OBJECTIVE
Study aims were to evaluate the safety and efficacy of the Food and Drug Administration-approved drug Vorinostat [suberoylanilide hydroxamic acid (SAHA)] in the treatment of canine corneal fibrosis using an in vitro model.


METHODS
Healthy donor canine corneas were collected and used to generate primary canine corneal fibroblasts (CCFs) by growing cultures in minimal essential medium supplemented with 10% fetal bovine serum. Canine corneal myofibroblasts, used as a model for corneal fibrosis, were produced by growing CCF cultures in serum-free medium containing transforming growth factor β1 (1 ng/mL). Trypan blue exclusion assays were used to determine the optimal SAHA dose for this in vitro model. Four hour after culturing with TGFβ1, CCF cultures were treated with 0.06% SAHA for 5 min (group 1) and for 24 h (group 2), representing single and multiple dose treatment regimes, respectively. Cultures were then further incubated in the presence of TGFβ1 (1 ng/μL) under serum-free conditions until they reached 70% confluence. Trypan blue exclusion, immunocytochemistry, and TUNEL assays were used to evaluate the cytotoxicity of SAHA. Real-time PCR, western blot analysis, and immunocytochemistry were used to determine the efficacy of SAHA to inhibit canine corneal myofibroblast formation.


RESULTS
Topical SAHA application in both treatment groups successfully decreased α-smooth muscle actin expression when compared to the TGFβ1 only treatment group (P < 0.05). Tested SAHA did not affect CCF phenotype or cellular viability and did not cause significant cell death.


CONCLUSIONS
Suberoylanilide hydroxamic acid safely and effectively inhibits TGFβ1-induced CCFs transformation to myofibroblast in vitro.
Harry S. Truman Veterans Memorial Hospital, Columbia, MO, USA.
Ann P
AP
Bosiack
Elizabeth A
EA
Giuliano
Rangan
R
Gupta
Rajiv R
RR
Mohan
D000818:Animals; D016923:Cell Death; D002478:Cells, Cultured; D003316:Corneal Diseases / Q000188:drug therapy / Q000662:veterinary*; D004285:Dogs; D005347:Fibroblasts / Q000187:drug effects*; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology / Q000592:standards / Q000627:therapeutic use; D006877:Hydroxamic Acids / Q000494:pharmacology* / Q000592:standards / Q000627:therapeutic use; D058628:Myofibroblasts / Q000187:drug effects; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology; D000077337:Vorinostat
D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D053773:Transforming Growth Factor beta1; D000077337:Vorinostat