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Updated: Sep 29, 2026
Characterization of In Vitro Differentiation of Human Primary Keratinocytes by RNA-Seq Analysis
Published on: May 16, 2020
A simple technique for high-throughput screening of drugs that modulate normal and psoriasis-like differentiation in
Arno Pol1, Mieke Bergers, Fred van Ruissen
1Department of Dermatology, University Hospital, Nijmegen, The Netherlands. a.pol@derma.azn.nl
Abstract:
Established treatments for psoriasis act ei-ther on hyperproliferation, inflammation, aberrant epidermal differentiation or a combination of these aspects of the disease. Potential new drugs for treatment of psoriasis or other disorders with abnormalities 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 cytokeratin 10 (CK10) and skin-derived antileucoprotease (SKALP)/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 psoriasis. All-trans-retinoic acid, dithranol and the p38 mitogen-activated protein (MAP) kinase inhibitor SB220025 displayed a strong inhibitory effect on SKALP expression while cyclosporin A, dexamethasone, the vitamin D(3) derivative calcipotriol and the p38 MAP kinase inhibitor SB203580 showed only moderate inhibition. Methotrexate and dimethylfumarate did not affect the expression of SKALP. With respect to CK10 expression, all-trans-retinoic acid, calcipotriol, SB203580 and SB220025 exhibited strong inhibition while dithranol showed only moderate suppression of this normal differentiation marker. Expression levels of CK10 were not significantly affected by dexamethasone, methotrexate, cyclosporin A or dimethylfumarate. 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 p38 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.
Insights
A new high-throughput screening model uses keratinocyte cultures to identify psoriasis drugs. This model effectively detects compounds that modify epidermal differentiation, including potent p38 MAP kinase inhibitors.
Area of Science:
- Dermatology and Pharmaceutical Sciences
- Cell Biology and Molecular Medicine
Background:
- Psoriasis treatments target hyperproliferation, inflammation, and aberrant epidermal differentiation.
- Identifying novel drugs for psoriasis and epidermal differentiation disorders requires efficient screening methods.
- Surrogate markers in keratinocyte models can facilitate high-throughput screening of compound libraries.
Purpose of the Study:
- To develop and validate a 96-well keratinocyte-based screening model for identifying pharmacologically active drugs.
- To utilize cytokeratin 10 (CK10) and skin-derived antileucoprotease (SKALP)/elafin as markers for normal and psoriatic epidermal differentiation.
- To assess the efficacy of known and experimental antipsoriatic drugs within this novel screening system.
Main Methods:
- Established two keratinocyte culture models in 96-well plates.
- Used CK10 and SKALP/elafin expression as markers for normal and psoriatic differentiation, respectively.
- Screened various compounds, including known antipsoriatic drugs and p38 MAP kinase inhibitors, at concentrations from 10(-7) to 10(-5) M.
Main Results:
- All-trans-retinoic acid, dithranol, and p38 MAP kinase inhibitor SB220025 strongly inhibited SKALP expression.
- All-trans-retinoic acid, calcipotriol, SB203580, and SB220025 strongly inhibited CK10 expression.
- The model identified p38 MAP kinase inhibitors as potent suppressors of differentiation-associated gene expression.
Conclusions:
- The developed screening model effectively parallels in vitro effects of known antipsoriatic drugs on keratinocytes.
- The system is suitable for high-throughput screening of compounds that modify epidermal differentiation.
- Further validation is needed, but the model shows promise for drug discovery in differentiation-related disorders.
