title: Skin retinoid concentrations are modulated by CYP26AI expression restricted to basal keratinocytes in normal human skin and differentiated 3D skin models.
abstract: Cellular levels of all-trans retinoic acid (RA) are meticulously regulated utilizing an array of systems to balance uptake, biosynthesis, catabolism, and efflux transport. Metabolic transformation of all-trans RA to 4-hydroxylated RA appears to be primarily catalyzed by the cytochrome P450 (CYP) 26AI. Analysis of monolayer cultures of normal human epidermal keratinocytes (NHEKs) and dermal fibroblasts by quantitative real-time PCR and reverse transcription-PCR revealed no basal levels of CYP26AI mRNA expression, whereas specific transcripts were detectable following addition of 10(-6) M all-trans RA. Immunofluorescence and Western blot analysis showed a weak expression of CYP26AI in NHEK, which was increased by stimulation with all-trans RA. Using a newly developed peptide antibody, we further examined the localization of CYP26AI expression in normal skin and three-dimensional (3D) skin models. In contrast to cell culture monolayers where CYP26AI was only weakly detectable, strong constitutive expression of CYP26AI in vivo and in organotypic culture was found to be restricted to basal epidermal keratinocytes, as well as eccrine sweat glands and sebaceous glands. These studies verify the capacity of human skin to metabolize RA, although substantial differences exist in CYP expression between normal skin and 3D skin models compared to monolayer cultures. Complex metabolic processes that maintain retinoid homeostasis may therefore be better studied in model systems more closely resembling in vivo skin. In light of our prior studies documenting the functional activity of RA metabolites, expression of CYP26 in the sebaceous gland epithelium supports the suggestion that altered RA metabolism may be involved in the pathogenesis of acne.
authors.affiliation: Department of Dermatology and Allergology, University Hospital of the RWTH Aachen, Aachen, Germany.
authors.forename: Ruth
authors.initials: R
authors.lastname: Heise
authors.forename: Jörg
authors.initials: J
authors.lastname: Mey
authors.forename: Mark M
authors.initials: MM
authors.lastname: Neis
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Marquardt
authors.forename: Sylvia
authors.initials: S
authors.lastname: Joussen
authors.forename: Hagen
authors.initials: H
authors.lastname: Ott
authors.forename: Tonio
authors.initials: T
authors.lastname: Wiederholt
authors.forename: Peter
authors.initials: P
authors.lastname: Kurschat
authors.forename: Mosaad
authors.initials: M
authors.lastname: Megahed
authors.forename: David R
authors.initials: DR
authors.lastname: Bickers
authors.forename: Hans F
authors.initials: HF
authors.lastname: Merk
authors.forename: Jens M
authors.initials: JM
authors.lastname: Baron
mesh_terms: D015153:Blotting, Western; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D004817:Epidermis / Q000201:enzymology; D005347:Fibroblasts / Q000201:enzymology; D005455:Fluorescent Antibody Technique; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects / Q000201:enzymology*; D009924:Organ Culture Techniques; D011565:Psoriasis / Q000201:enzymology; D012333:RNA, Messenger / Q000378:metabolism; D000072516:Retinoic Acid 4-Hydroxylase; D012176:Retinoids / Q000032:analysis / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012627:Sebaceous Glands / Q000201:enzymology; D012867:Skin / Q000187:drug effects / Q000201:enzymology*; D014801:Vitamin A / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D012333:RNA, Messenger; D012176:Retinoids; D014801:Vitamin A; D003577:Cytochrome P-450 Enzyme System; D000072516:Retinoic Acid 4-Hydroxylase