A simple technique for high-throughput screening of drugs that modulate normal and XXXX -like differentiation in cultured human keratinocytes.

@entity70950 @entity32 @entity13959 @entity1569 @entity66 @entity3875 @entity1441 @entity205 @entity7141 @entity619 @entity15626 @entity1435 @entity2116 @entity1499 @entity1968 @entity7434
Established treatments for @entity1569 act ei-ther on hyperproliferation, @entity32 , aberrant epidermal differentiation or a combination of these aspects of the disease. Potential new drugs for treatment of @entity1569 or other @entity66 in epidermal differentiation can be identified by high-throughput screening of large compound libraries using surrogate markers for the disease. Here we describe a screening model to detect pharmacologically active drugs in two keratinocyte-based, 96-well culture models that use expression of @entity7434 ( @entity7434 ) and @entity13959 ( @entity13959 )/elafin as markers for normal and psoriatic differentiation, respectively, and allow multiple parameters to be determined from a single well. In this model we tested a number of compounds in a pharmacological range from 10(-7) to 10(-5) M, including known antipsoriatic drugs, and experimental drugs that are potentially useful in the treatment of @entity1569 . @entity1968 , @entity7141 and the @entity1435 mitogen-activated protein (MAP) kinase inhibitor @entity70950 displayed a strong inhibitory effect on @entity13959 expression while @entity205 , @entity619 , the @entity3875 derivative @entity2116 and the @entity1435 MAP kinase inhibitor @entity1441 showed only moderate inhibition. @entity1499 and @entity15626 did not affect the expression of @entity13959 . With respect to @entity7434 expression, @entity1968 , @entity2116 , @entity1441 and @entity70950 exhibited strong inhibition while @entity7141 showed only moderate suppression of this normal differentiation marker. Expression levels of @entity7434 were not significantly affected by @entity619 , @entity1499 , @entity205 or @entity15626 . This model system parallels most, but not all, findings on the in vitro effect of known antipsoriatic drugs on keratinocytes. In addition, the model identifies @entity1435 MAP kinase inhibitors as potent suppressors of differentiation-associated gene expression. Although further delineation and validation of this model is required, we conclude that the system is amenable to down-scaling and application as a high-throughput screen for differentiation-modifying compounds.
