THE RETINOID-X RECEPTOR AGONIST 9-CIS-RETINOIC ACID AND THE 24-HYDROXYLASE INHIBITOR KETOCONAZOLE INCREASE ACTIVITY OF 1,25-DIHYDROXYVITAMIN D-3 IN HUMAN SKIN IN-VIVO

1,25-Dihydroxyvitamin D-3 [1,25(OH)(2)D-3] transactivates its target g enes via the vitamin D receptor (VDR), VDR functions in physiology as a dimer complexed with retinoid X receptor (RXR), whose natural ligand is 9-cis-retinoic acid (9-c-RA), Inactivation of 1,25(OH)(2)D-3 occur s through a cytochrome P-450 24-hydroxylase (OHase). The promoter of t he human 24-OHase gene contains a 1,25(OH)(2)D-3-responsive enhancer e lement (VDRE). We have used this VDRE containing gene as an endogenous reporter for vitamin D-3-mediated gene activation in vivo, Normal adu lt human skin was keratomed after a 2-d exposure to 1,25(OH)(2)D-3, 9- c-RA, all-trans-RA, and ketoconazole, 1,25(OH)(2)D-3 caused a concentr ation-dependent increase in 24-OHase mRNA expression as determined by northern blot analysis, The activity of epidermal 24-OHase was also in duced by 1,25(OH)(2)D-3. Compared with vehicle, neither of the RA isom ers nor ketoconazole alone induced 24-OHase mRNA. Addition of 9-c-RA o r t-RA to 1,25(OH)(2)D-3, however, caused a synergistic increase in 24 -OHase mRNA, Similarly, 1,25(OH)(2)D-3 plus ketoconazole increased 24- OHase mRNA synergistically. Ketoconazole inhibited ex vivo 1,25(OH)(2) D-3-induced epidermal 24-OHase activity, Thus, 24-OHase mRNA induction is a sensitive reporter of 1,25(OH)(2)D-3 activity in vivo; RXR bound to VDR is not a silent partner in vivo, because 9-c-RA enhances 1,25( OH)(2)D-3-liganded RXRNDR stimulation of the VDRE containing 24-OHase gene; ketoconazole inhibition of 24-OHase enhances 1,25(OH)(2)D-3 acti vity by impeding its breakdown, Thus, the synergistic response of huma n skin to topical 1,25(OH)(2)D-3 and/or 1,25(OH),D, analogs plus RXR r etinoids and/or ketoconazole may be exploited to give a desired biolog ic/therapeutic response with less 1,25(OH)(2)D-3, minimizing the poten tial calcemic risk from systemic absorption of 1,25(OH)(2)D-3.