Cystatin M/E knockdown by lentiviral delivery of shRNA impairs epidermal morphogenesis of human skin equivalents

• Published in: Experimental dermatology 2012-11, Vol.21 (11), p.889-891
• Main Authors: Jansen, Patrick A. M., van den Bogaard, Ellen H., Kersten, Ferry F. J., Oostendorp, Corien, van Vlijmen-Willems, Ivonne M. J. J., Oji, Vinzenz, Traupe, Heiko, Hennies, Hans C., Schalkwijk, Joost, Zeeuwen, Patrick L. J. M.
• Format: Article
• Language: English
• Subjects: Animal models; Biological and medical sciences; Cell Proliferation - drug effects; Cells, Cultured; Cystatin M - genetics; Cystatin M - physiology; Dermatology; Dyskeratosis; Epidermis - cytology; Epidermis - drug effects; Epidermis - growth & development; Gene Knockdown Techniques; human skin equivalents; Humans; ichthyosis; Ichthyosis - pathology; lentivirus; Lentivirus - genetics; Medical sciences; Models, Biological; Morphogenesis - drug effects; Morphogenesis - genetics; Phenotype; protease inhibitor; RNA, Small Interfering - genetics; RNA, Small Interfering - pharmacology; short hairpin RNA; Skin, Artificial; Tissue Scaffolds
• ISSN: 0906-6705; 1600-0625
• DOI: 10.1111/exd.12022

The protease inhibitor cystatin M/E (CST6) regulates a biochemical pathway involved in stratum corneum homeostasis, and its deficiency in mice causes ichthyosis and neonatal lethality. Cystatin M/E deficiency has not been described in humans so far, and we did not detect disease‐causing mutations in the CST6 gene in a large number of patients with autosomal recessive congenital ichthyosis, who were negative for mutations in known ichthyosis‐associated genes. To investigate the phenotype of CST6 deficiency in human epidermis, we used lentiviral delivery of short hairpin RNAs that target CST6 in a 3D reconstructed skin model. Surprisingly, CST6 deficiency did not cause an ichthyosis‐like phenotype, but prevented the development of a multilayered epidermis. From this study, we conclude that CST6 deficiency may be incompatible with normal human foetal development.