Abstract
The biosynthesis of retinoic acid (RA) from retinol is controlled by several enzymes, e.g. dehydrogenases (RalDH2, RoDH-4) and retinol-esterifying enzyme (LRAT), whereas its degradation mainly involves CYP26 enzymes. In keratinocytes, RA activates the nuclear retinoid-receptors inducing the transcription of many genes. Here, we examined the effects of RA and the CYP26 inhibitors, liarozole and talarozole, on retinoid metabolism and RA-regulated genes in organotypic epidermis. RA induced the expression of CYP26 enzymes already after 8 h, whereas LRAT exhibited a later response and peaked at 48 h, indicating a feedback induction of retinol esterification. In line with a reduced biosynthesis of RA from retinol after exogenous RA, the expression of RDH16 reduced 80% in response to exogenous RA. The mRNA expression of RA-regulated genes (KRT2, KRT4, CRABPII and HBEGF) was altered within 24 h after RA exposure. In contrast, the CYP26 inhibitors caused only minor effects, except for a clear-cut induction of CYP26A1 only when combined with minute amounts of exogenous RA. Cellular accumulation of exogenous [3H]RA was higher after talarozole than after liarozole, probably indicating a greater CYP26-inhibitory potency of the former drug. The present study shows that CYP26A1 expression is extremely sensitive to both exogenous RA and increased endogenous RA levels, i.e. due to CYP26 inhibition, and thus an excellent biomarker for retinoid signalling in organotypic epidermis.
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Abu-Abed S, MacLean G, Fraulob V, Chambon P, Petkovich M, Dolle P (2002) Differential expression of the retinoic acid-metabolizing enzymes CYP26A1 and CYP26B1 during murine organogenesis. Mech Dev 110:173–177. doi:10.1016/S0925-4773(01)00572-X
Allenby G, Bocquel MT, Saunders M, Kazmer S, Speck J, Rosenberger M, Lovey A, Kastner P, Grippo JF, Chambon P, Levin AA (1993) Retinoic acid receptors and retinoid X-receptors: interactions with endogenous retinoic acids. Proc Natl Acad Sci USA 90:30–34. doi:10.1073/pnas.90.1.30
Åström A, Pettersson U, Chambon P, Voorhees J (1994) Retinoic acid induction of human cellular retinoic acid-binding protein-II gene transcription is mediated by retinoic acid receptor–retinoid X receptor heterodimers bound to one far upstream retinoic acid-response element with 5-base pair spacing. J Biol Chem 269:22334–22339
Bernard FX, Pedretti N, Rosdy M, Deguercy A (2002) Comparison of gene expression profiles in human keratinocyte mono-layer cultures, reconstituted epidermis and normal human skin: transcriptional effects of retinoid treatments in reconstituted human epidermis. Exp Dermatol 11:59–74. doi:10.1034/j.1600-0625.2002.110107.x
Blomhoff R, Blomhoff H (2007) From carotenoids and Vitamin A to retinoids. In: Vahlquist A, Duvic M (eds) Retinoids and carotenoids in dermatology. Informa Healthcare USA Inc., New York, pp 1–26
Bovenschen HJ, Otero ME, Langewouters AM, van Vlijmen-Willems IM, van Rens DW, Seyger MM, van de Kerkhof PC (2007) Oral retinoic acid metabolism blocking agent Rambazole for plaque psoriasis: an immunohistochemical study. Br J Dermatol 156:263–270. doi:10.1111/j.1365-2133.2006.07660.x
Duester G (2000) Families of retinoid dehydrogenases regulating vitamin A function production of visual pigment and retinoic acid. Eur J Biochem 267:4315–4324. doi:10.1046/j.1432-1327.2000.01497.x
Elder J, Cromie M, Griffiths C, Chambon P, Voorhees J (1993) Stimulus-selective induction of CRABP-II mRNA: a marker for retinoic acid action in human skin. J Invest Dermatol 100:356–359. doi:10.1111/1523-1747.ep12471816
Fisher GJ, Esmann J, Griffiths CEM, Talwar HS, Duell EA, Hammerberg C, Elder JT, Karabin GD, Nickoloff BJ, Cooper KD, Voorhees JJ (1991) Cellular, immunologic and biochemical characterization of topical retinoic acid-treated human skin. J Invest Dermatol 96:699–707. doi:10.1111/1523-1747.ep12470632
Garzon R, Pichiorri F, Palumbo T, Visentini M, Aqeilan R, Cimmino A, Wang H, Sun H, Volinia S, Alder H, Calin GA, Liu CG, Andreeff M, Croce CM (2007) MicroRNA gene expression during retinoic acid-induced differentiation of human acute promyelocytic leukemia. Oncogene 26:4148–4157. doi:10.1038/sj.onc.1210186
Gaub M-P, Lutz Y, Ghyselinck NB, Scheuer I, Pfister V, Chambon P, Rochette-Egly C (1998) Nuclear detection of cellular retinoic acid-binding proteins I and II with new antibodies. J Histochem Cytochem 46:1103–1111
Karlsson T, Virtanen M, Sirsjö A, Rollman O, Vahlquist A, Törmä H (2002) Topical retinoic acid alters the expression of cellular retinoic acid-binding protein-I and cellular retinoic acid-binding protein-II in non-lesional but not lesional psoriatic skin. Exp Dermatol 11:143–152. doi:10.1034/j.1600-0625.2002.110206.x
Kuijpers AL, Van Pelt JP, Bergers M, Boegheim PJ, Den Bakker JE, Siegenthaler G, Van de Kerkhof PC, Schalkwijk J (1998) The effects of oral liarozole on epidermal proliferation and differentiation in severe plaque psoriasis are comparable with those of acitretin. Br J Dermatol 139:380–389. doi:10.1046/j.1365-2133.1998.02399.x
Kurlandsky SB, Duell EA, Kang S, Voorhees JJ, Fisher GJ (1996) Auto-regulation of retinoic acid biosynthesis through regulation of retinol esterification in human keratinocytes. J Biol Chem 271:15346–15352. doi:10.1074/jbc.271.26.15346
Loudig O, Babichuk C, White J, Abu-Abed S, Mueller C, Petkovich M (2000) Cytochrome P450RAI(CYP26) promoter: a distinct composite retinoic acid response element underlies the complex regulation of retinoic acid metabolism. Mol Endocrinol 14:1483–1497. doi:10.1210/me.14.9.1483
Loudig O, Maclean GA, Dore NL, Luu L, Petkovich M (2005) Transcriptional co-operativity between distant retinoic acid response elements in regulation of Cyp26A1 inducibility. Biochem J 392:241–248. doi:10.1042/BJ20050874
Lucker GP, Heremans AM, Boegheim PJ, van de Kerkhof PC, Steijlen PM (1997) Oral treatment of ichthyosis by the cytochrome P-450 inhibitor liarozole. Br J Dermatol 136:71–75. doi:10.1111/j.1365-2133.1997.tb08749.x
Lucker GP, Verfaille CJ, Heremans AM, Vanhoutte FP, Boegheim JP, Steijlen PP (2005) Topical liarozole in ichthyosis: a double-blind, left–right comparative study followed by a long-term open maintenance study. Br J Dermatol 152:566–569. doi:10.1111/j.1365-2133.2005.06399.x
Napoli JL (1999) Interactions of retinoid-binding proteins and enzymes in retinoid metabolism. Biochim Biophys Acta 1440:139–162
Noy N (2000) Retinoid-binding proteins: mediators of retinoid action. Biochem J 348(Pt 3):481–495. doi:10.1042/0264-6021:3480481
Pavez Loriè E, Cools M, Borgers M, Wouters L, Shroot B, Hagforsen E, Törmä H, Vahlquist A (2009) Topical treatment with CYP26 inhibitor talarozole (R115866) dose dependently alters the expression of retinoid-regulated genes in normal human epidermis. Br J Dermatol 160:26–36. doi:10.1111/j.1365-2133.2008.08895.x
Pavez Loriè E, Gånemo A, Borgers M, Wouters L, Blockhuys S, vandePlassche L, Törmä H, Vahlquist A (2009) Expression of retinoid-regulated genes in lamellar ichthyosis (LI) vs. healthy control epidermis: changes after oral treatment with liarozole. Acta Derm Venereol 89:12–20
Poumay Y, Dupont F, Marcoux S, Leclercq-Smekens M, Herin M, Coquette A (2004) A simple reconstructed human epidermis: preparation of the culture model and utilization in in vitro studies. Arch Dermatol Res 296:203–211. doi:10.1007/s00403-004-0507-y
Roos TC, Jugert FK, Merk HF, Bickers DR (1998) Retinoid metabolism in the skin. Pharmacol Rev 50:315–333
Rosdy M, Bertino B, Butet V, Gibbs S, Ponec M, Darmon M (1997) Retinoic acid inhibits epidermal differentiation when applied topically on the stratum corneum of epidermis formed in vitro by human keratinocytes grown on defined medium. In Vitro Toxicol 10:39–47
Saurat JH, Didierjean L, Masgrau E, Piletta PA, Jaconi S, Chatellardgruaz D, Gumowski D, Masouye I, Salomon D, Siegenthaler G (1994) Topical retinaldehyde on human skin: biologic effects and tolerance. J Invest Dermatol 103:770–774. doi:10.1111/1523-1747.ep12412861
Stoll SW, Elder JT (1998) Retinoid regulation of heparin-binding EGF-like growth factor gene expression in human keratinocytes and skin. Exp Dermatol 7:391–397. doi:10.1111/j.1600-0625.1998.tb00339.x
Taimi M, Helvig C, Wisniewski J, Ramshaw H, White J, Amad M, Korczak B, Petkovich M (2004) A novel human cytochrome P450, CYP26C1, involved in metabolism of 9-cis and all-trans isomers of retinoic acid. J Biol Chem 279:77–85. doi:10.1074/jbc.M308337200
Tay YM, Tam WL, Ang YS, Gaughwin PM, Yang H, Wang W, Liu R, George J, Ng HH, Perera RJ, Lufkin T, Rigoutsos I, Thomson AM, Lim B (2008) MicroRNA-134 modulates the differentiation of mouse embryonic stem cells, where it causes post-transcriptional attenuation of Nanog and LRH1. Stem Cell 26:17–29. doi:10.1634/stemcells.2007-0295
Törmä H, Rollman O, Vahlquist A (1999) The vitamin A metabolism and expression of retinoid-binding proteins differ in HaCaT cells and normal human keratinocytes. Arch Dermatol Res 291:339–345. doi:10.1007/s004030050419
Törmä H, Geijer S, Gester T, Alpholm K, Berne B, Lindberg M (2006) Variations in the mRNA expression of inflammatory mediators, markers of differentiation and lipid-metabolizing enzymes caused by sodium lauryl sulphate in cultured human keratinocytes. Toxicol In Vitro 20:472–479. doi:10.1016/j.tiv.2005.09.011
van de Kerkhof PC (2006) Update on retinoid therapy of psoriasis in: an update on the use of retinoids in dermatology. Dermatol Ther 19:252–263. doi:10.1111/j.1529-8019.2006.00082.x
Verfaille CJ, Coel M, Boersma IH, Mertens J, Borgers M, Roseeuw D (2007) Oral R115866 in the treatment of moderate to severe facial acne vulgaris: an exploratory study. Br J Dermatol 157:122–126. doi:10.1111/j.1365-2133.2007.07896.x
Verfaille CJ, Vanhoutte FP, Blanchet-Bardon C, van Steensel MA, Steijlen PM (2007) Oral liarozole vs. acitretin in the treatment of ichthyosis: a phase II/III multicentre, double-blind, randomized, active-controlled study. Br J Dermatol 156:965–973. doi:10.1111/j.1365-2133.2006.07745.x
Verfaille CJ, Borgers M, van Steensel MA (2008) Retinoic acid metabolism blocking agents (RAMBAs): a new paradigm in the treatment of hyperkeratotic disorders. J Dtsch Dermatol Ges 6:355–364. doi:10.1111/j.1610-0387.2007.06541.x
Virtanen M (2001) On keratin mutations in epidermolytic hyperkeratosis and the regulation of keratin expression by retinoids. Comprehensive summaries of Uppsala dissertations from the Faculty of Medicine 1051, Uppsala University, Uppsala
Virtanen M, Törmä H, Vahlquist A (2000) Keratin 4 upregulation by retinoic acid in vivo: a sensitive marker for retinoid bioactivity in human epidermis. J Invest Dermatol 114:487–493. doi:10.1046/j.1523-1747.2000.00901.x
Welch C, Chen Y, Stallings RL (2007) MicroRNA-34a functions as a potential tumor suppressor by inducing apoptosis in neuroblastoma cells. Oncogene 26:5017–5022. doi:10.1038/sj.onc.1210293
White JA, Guo YD, Baetz K, Beckett-Jones B, Bonasoro J, Hsu KE, Dilworth FJ, Jones G, Petkovich M (1996) Identification of the retinoic acid-inducible all-trans-retinoic acid 4-hydroxylase. J Biol Chem 271:29922–29927. doi:10.1074/jbc.271.47.29922
White JA, Ramshaw H, Taimi M, Stangle W, Zhang A, Everingham S, Creighton S, Tam SP, Jones G, Petkovich M (2000) Identification of the human cytochrome P450, P450RAI-2, which is predominantly expressed in the adult cerebellum and is responsible for all-trans-retinoic acid metabolism. Proc Natl Acad Sci USA 97:6403–6408. doi:10.1073/pnas.120161397
White RJ, Nie Q, Lander AD, Schilling TF (2007) Complex regulation of cyp26a1 creates a robust retinoic acid gradient in the zebrafish embryo. PLoS Biol 5:e304. doi:10.1371/journal.pbio.0050304
Yoshimura K, Uchida G, Okazaki M, Kitano Y, Harii K (2003) Differential expression of heparin-binding EGF-like growth factor (HB-EGF) mRNA in normal human keratinocytes induced by a variety of natural and synthetic retinoids. Exp Dermatol 12(Suppl 2):28–34. doi:10.1034/j.1600-0625.12.s2.5.x
Acknowledgments
The valuable technical assistance by Inger Pihl-Lundin is gratefully acknowledged. This work was funded by grants from the Swedish Research Council (74X-07133), the Erik, Karin och Gösta Selanders Foundation, the Edvard Welander and Finsen Foundations and The Swedish Psoriasis Foundation.
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Pavez Loriè, E., Chamcheu, J.C., Vahlquist, A. et al. Both all-trans retinoic acid and cytochrome P450 (CYP26) inhibitors affect the expression of vitamin A metabolizing enzymes and retinoid biomarkers in organotypic epidermis. Arch Dermatol Res 301, 475–485 (2009). https://doi.org/10.1007/s00403-009-0937-7
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DOI: https://doi.org/10.1007/s00403-009-0937-7