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Identifying targets for topical RNAi therapeutics in psoriasis: assessment of a new in vitro psoriasis model

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Abstract

Diseases of the skin are amenable to RNAi-based therapies and targeting key components in the pathophysiology of psoriasis using RNAi may represent a successful new therapeutic strategy. We aimed to develop a straightforward and highly reproducible in vitro psoriasis model useful to study the effects of gene knockdown by RNAi and to identify new targets for topical RNAi therapeutics. We evaluated the use of keratinocytes derived from psoriatic plaques and normal human keratinocytes (NHKs). To induce a psoriatic phenotype in NHKs, combinations of pro-inflammatory cytokines (IL-1α, IL-17A, IL-6 and TNF-α) were tested. The model based on NHK met our needs of a reliable and predictive preclinical model, and this model was further selected for gene expression analyses, comprising a panel of 55 psoriasis-associated genes and five micro-RNAs (miRNAs). Gene silencing studies were conducted by using small interfering RNAs (siRNAs) and miRNA inhibitors directed against potential target genes such as CAMP and DEFB4 and miRNAs such as miR-203. We describe a robust and highly reproducible in vitro psoriasis model that recapitulates expression of a large panel of genes and miRNAs relevant to the pathogenesis of psoriasis. Furthermore, we show that our model is a powerful first step model system for testing and screening RNAi-based therapeutics.

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Acknowledgments

We thank Martine De Mil for help with cell culture, and Marie-Chantal Herteleer and Els Van Maelsaeke for technical assistance. Dr. S. Bracke is funded by an IWT grant (091208) (‘Flemish government agency for Innovation by Science and Technology’).

Conflict of interest

None declared.

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Authors and Affiliations

  1. Department of Dermatology 2K4, Ghent University Hospital, De Pintelaan 185, 9000, Ghent, Belgium

    S. Bracke, E. Desmet, M. Van Gele & J. Lambert

  2. Regenerative Medicine Unit and the Cutaneous Diseases Modeling Unit, Medioambientales y Tecnológicas (CIEMAT) and Centre for Biomedical Research on Rare Diseases (CIBERER), Madrid, Spain

    S. Guerrero-Aspizua

  3. Epithelial Biomedicine Division, Basic Research Department, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT) and Centre for Biomedical Research on Rare Diseases (CIBERER), Madrid, Spain

    M. Carretero

  4. Department of Dermatology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands

    S. G. Tjabringa & J. Schalkwijk

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  1. S. Bracke
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  2. E. Desmet
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Correspondence to S. Bracke.

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Bracke, S., Desmet, E., Guerrero-Aspizua, S. et al. Identifying targets for topical RNAi therapeutics in psoriasis: assessment of a new in vitro psoriasis model. Arch Dermatol Res 305, 501–512 (2013). https://doi.org/10.1007/s00403-013-1379-9

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  • DOI: https://doi.org/10.1007/s00403-013-1379-9

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