TY - JOUR
T1 - Characterization of a new pathway that activates lumisterol in vivo to biologically active hydroxylumisterols
AU - Slominski, Andrzej T.
AU - Kim, Tae-Kang
AU - Hobrath, Judith V.
AU - Janjetovic, Zorica
AU - Oak, Allen S. W.
AU - Postlethwaite, Arnold
AU - Lin, Zongtao
AU - Li, Wei
AU - Takeda, Yukimasa
AU - Jetten, Anton M.
AU - Tuckey, Robert C.
PY - 2017/12/1
Y1 - 2017/12/1
N2 - Using LC/qTOF-MS we detected lumisterol, 20-hydroxylumisterol, 22-hydroxylumisterol, 24-hydroxylumisterol, 20,22-dihydroxylumisterol, pregnalumisterol, 17-hydroxypregnalumisterol and 17,20-dihydroxypregnalumisterol in human serum and epidermis, and the porcine adrenal gland. The hydroxylumisterols inhibited proliferation of human skin cells in a cell type-dependent fashion with predominant effects on epidermal keratinocytes. They also inhibited melanoma proliferation in both monolayer and soft agar. 20-Hydroxylumisterol stimulated the expression of several genes, including those associated with keratinocyte differentiation and antioxidative responses, while inhibiting the expression of others including RORA and RORC. Molecular modeling and studies on VDRE-transcriptional activity excludes action through the genomic site of the VDR. However, their favorable interactions with the A-pocket in conjunction with VDR translocation studies suggest they may act on this non-genomic VDR site. Inhibition of ROR alpha and ROR gamma transactivation activities in a Tet-on CHO cell reporter system, ROR alpha co-activator assays and inhibition of (RORE)-LUC reporter activity in skin cells, in conjunction with molecular modeling, identified ROR alpha and ROR gamma as excellent receptor candidates for the hydroxylumisterols. Thus, we have discovered a new biologically relevant, lumisterogenic pathway, the metabolites of which display biological activity. This opens a new area of endocrine research on the effects of the hydroxylumisterols on different pathways in different cells and the mechanisms involved.
AB - Using LC/qTOF-MS we detected lumisterol, 20-hydroxylumisterol, 22-hydroxylumisterol, 24-hydroxylumisterol, 20,22-dihydroxylumisterol, pregnalumisterol, 17-hydroxypregnalumisterol and 17,20-dihydroxypregnalumisterol in human serum and epidermis, and the porcine adrenal gland. The hydroxylumisterols inhibited proliferation of human skin cells in a cell type-dependent fashion with predominant effects on epidermal keratinocytes. They also inhibited melanoma proliferation in both monolayer and soft agar. 20-Hydroxylumisterol stimulated the expression of several genes, including those associated with keratinocyte differentiation and antioxidative responses, while inhibiting the expression of others including RORA and RORC. Molecular modeling and studies on VDRE-transcriptional activity excludes action through the genomic site of the VDR. However, their favorable interactions with the A-pocket in conjunction with VDR translocation studies suggest they may act on this non-genomic VDR site. Inhibition of ROR alpha and ROR gamma transactivation activities in a Tet-on CHO cell reporter system, ROR alpha co-activator assays and inhibition of (RORE)-LUC reporter activity in skin cells, in conjunction with molecular modeling, identified ROR alpha and ROR gamma as excellent receptor candidates for the hydroxylumisterols. Thus, we have discovered a new biologically relevant, lumisterogenic pathway, the metabolites of which display biological activity. This opens a new area of endocrine research on the effects of the hydroxylumisterols on different pathways in different cells and the mechanisms involved.
KW - CYTOCHROME P450SCC CYP11A1
KW - PITUITARY-ADRENAL AXIS
KW - BASAL-CELL CARCINOMA
KW - VITAMIN-D METABOLISM
KW - KAPPA-B ACTIVITY
KW - EPIDERMAL-KERATINOCYTES
KW - ULTRAVIOLET-RADIATION
KW - MELANIN PIGMENTATION
KW - MAMMALIAN SKIN
KW - D ANALOGS
U2 - 10.1038/s41598-017-10202-7
DO - 10.1038/s41598-017-10202-7
M3 - Article
C2 - 28900196
SN - 2045-2322
VL - 7
JO - Scientific Reports
JF - Scientific Reports
IS - 1
M1 - 11434
ER -