title: The use of rat lens explant cultures to study the mechanism of drug-induced cataractogenesis.
abstract: Lens explant cultures were used to assess the mechanism of drug-induced cataractogenic potential of NVS001, a peroxisome proliferator-activated receptor delta (PPARδ) agonist, which resulted in cataract in all treated animals during a 13-week rat study. Ciglitazone, a PPARγ agonist and cataractogenic compound, was used as a positive control to validate this model. Rat lenses were extracted and cultured in medium supplemented with antibiotics for 24-h preincubation pretreatment. Lenses showing no signs of damage at the end of the preincubation pretreatment period were randomized into five experimental groups, (1) untreated control, (2) 0.1% dimethyl sulphoxide control, (3) 10μM NVS001, (4) 10μM ciglitazone, and (5) 10μM acetaminophen (negative control). Lenses were treated every 24 h after preincubation pretreatment for up to 48 h. Samples for viability, histology, and gene expression profiling were collected at 4, 24, and 48 h. There was a time-dependent increase in opacity, which correlated to a decrease in viability measured by adenosine triphosphate levels in NVS001 and ciglitazone-treated lenses compared with controls. NVS001 and ciglitazone had comparable cataractogenic effects after 48 h with histology showing rupture of the lens capsule, lens fiber degeneration, cortical lens vacuolation, and lens epithelial degeneration. Furthermore, no changes were seen when lenses were treated with acetaminophen. Gene expression analysis supported oxidative and osmotic stress, along with decreases in membrane and epithelial cell integrity as key factors in NVS001-induced cataracts. This study suggests that in vitro lens cultures can be used to assess cataractogenic potential of PPAR agonists and to study/understand the underlying molecular mechanism of cataractogenesis in rat.
authors.affiliation: Investigative Toxicology, Novartis Institutes of Biomedical Research, East Hanover, New Jersey 07936, USA.
authors.forename: Shruthi
authors.initials: S
authors.lastname: Sampath
authors.forename: Lee Anne
authors.initials: LA
authors.lastname: McLean
authors.forename: Chiara
authors.initials: C
authors.lastname: Buono
authors.forename: Pierre
authors.initials: P
authors.lastname: Moulin
authors.forename: Armin
authors.initials: A
authors.lastname: Wolf
authors.forename: Salah-Dine
authors.initials: SD
authors.lastname: Chibout
authors.forename: Francois
authors.initials: F
authors.lastname: Pognan
authors.forename: Steve
authors.initials: S
authors.lastname: Busch
authors.forename: Nandita
authors.initials: N
authors.lastname: Shangari
authors.forename: Elba
authors.initials: E
authors.lastname: Cruz
authors.forename: Maya
authors.initials: M
authors.lastname: Gurnani
authors.forename: Parul
authors.initials: P
authors.lastname: Patel
authors.forename: Albert
authors.initials: A
authors.lastname: Reising
mesh_terms: D000085:Acetates / Q000494:pharmacology / Q000633:toxicity*; D000818:Animals; D002386:Cataract / Q000139:chemically induced* / Q000473:pathology; D002470:Cell Survival / Q000187:drug effects; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D007908:Lens, Crystalline / Q000187:drug effects* / Q000473:pathology; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis; D018384:Oxidative Stress / Q000187:drug effects; D047494:PPAR delta / Q000819:agonists*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013844:Thiazoles / Q000494:pharmacology / Q000633:toxicity*; D045162:Thiazolidinediones / Q000494:pharmacology / Q000633:toxicity*; D046509:Tissue Culture Techniques
chemical_list: C573693:2-(4-((4-(6-methoxy-3-pyridinyl)-5-(4-(trifluoromethoxy)phenyl)-2-thiazolyl)methoxy)-2-methylphenoxy)acetic acid; D000085:Acetates; D005136:Eye Proteins; D047494:PPAR delta; D012333:RNA, Messenger; D013844:Thiazoles; D045162:Thiazolidinediones; C039671:ciglitazone