RORo: And ROR 7 are expressed in human skin and serve as receptors for endogenously produced noncalcemic 20-hydroxy- and 20,23-dihydroxyvitamin D

A.T. Słomiński, T. Kim, Y. Takeda, Z. Janjetovic, A.A. Brozyna, C. Skobowiat, J. Wang, A.E. Postlethwaite, W. Li, Robert Tuckey, A.M. Jetten

Research output: Contribution to journalArticle

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Abstract

RORa and RORy are expressed in human skin cells that produce the noncalcemic 20-hydroxyvi-tamin D3 [20(OH)D3] and 20,23-dihydroxyvitamin D3 [20,23(OH)2D3]. Chinese hamster ovary (CHO) cells stably expressing a Tet-on RORa or RORy expression vector and a ROR-responsive element (RORE)-LUC reporter, and a mammalian 2-hybrid model examining the interaction between the ligand binding domain (LBD) of RORa or RORy with an LBD-interacting LXXLL-peptide, were used to study ROR-antagonist activities. These assays revealed that 20(OH)D3 and 20,23(OH)2D3 function as antagonists of RORa and RORy. Moreover, 20(OH)D3 inhibited the activation of the promoter of the Bmal1 and G6pase genes, targets of RORa, and 20(OH)D3 and 20,23(OH)2D3 inhibited Il17 promoter activity in Jurkat cells overexpressing RORa or RORy. Molecular modeling using crystal structures of the LBDs of RORa and RORy revealed docking scores for 20(OH)D3, 20, 23(OH)2D3 and 1,25(OH)2D3 similar to those of the natural ligands, predicting good binding to the receptor. Notably, 20(OH)D3, 20,23(OH)2D3, and 1,25(OH)2D3 inhibited RORE-mediated activation of a reporter in keratinocytes and melanoma cells and inhibited IL-17 production by immune cells. Our study identifies a novel signaling pathway, in which 20(OH)D3 and 20,23(OH)2D3 act as antagonists or inverse agonists of RORa and RORy, that opens new possibilities for local (skin) or systemic regulation. © FASEB.
Original languageEnglish
Pages (from-to)2775-2789
JournalFASEB Journal
Volume28
Issue number7
DOIs
Publication statusPublished - 2014

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Dihydroxycholecalciferols
Skin
Ligands
Chemical activation
Jurkat Cells
Molecular modeling
Interleukin-17
Cricetulus
20-hydroxyvitamin D3
20,23-dihydroxyvitamin D3
Keratinocytes
Ovary
Melanoma
Assays
Genes
Crystal structure
Cells
Peptides

Cite this

Słomiński, A. T., Kim, T., Takeda, Y., Janjetovic, Z., Brozyna, A. A., Skobowiat, C., ... Jetten, A. M. (2014). RORo: And ROR 7 are expressed in human skin and serve as receptors for endogenously produced noncalcemic 20-hydroxy- and 20,23-dihydroxyvitamin D. FASEB Journal, 28(7), 2775-2789. https://doi.org/10.1096/fj.13-242040
Słomiński, A.T. ; Kim, T. ; Takeda, Y. ; Janjetovic, Z. ; Brozyna, A.A. ; Skobowiat, C. ; Wang, J. ; Postlethwaite, A.E. ; Li, W. ; Tuckey, Robert ; Jetten, A.M. / RORo: And ROR 7 are expressed in human skin and serve as receptors for endogenously produced noncalcemic 20-hydroxy- and 20,23-dihydroxyvitamin D. In: FASEB Journal. 2014 ; Vol. 28, No. 7. pp. 2775-2789.
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abstract = "RORa and RORy are expressed in human skin cells that produce the noncalcemic 20-hydroxyvi-tamin D3 [20(OH)D3] and 20,23-dihydroxyvitamin D3 [20,23(OH)2D3]. Chinese hamster ovary (CHO) cells stably expressing a Tet-on RORa or RORy expression vector and a ROR-responsive element (RORE)-LUC reporter, and a mammalian 2-hybrid model examining the interaction between the ligand binding domain (LBD) of RORa or RORy with an LBD-interacting LXXLL-peptide, were used to study ROR-antagonist activities. These assays revealed that 20(OH)D3 and 20,23(OH)2D3 function as antagonists of RORa and RORy. Moreover, 20(OH)D3 inhibited the activation of the promoter of the Bmal1 and G6pase genes, targets of RORa, and 20(OH)D3 and 20,23(OH)2D3 inhibited Il17 promoter activity in Jurkat cells overexpressing RORa or RORy. Molecular modeling using crystal structures of the LBDs of RORa and RORy revealed docking scores for 20(OH)D3, 20, 23(OH)2D3 and 1,25(OH)2D3 similar to those of the natural ligands, predicting good binding to the receptor. Notably, 20(OH)D3, 20,23(OH)2D3, and 1,25(OH)2D3 inhibited RORE-mediated activation of a reporter in keratinocytes and melanoma cells and inhibited IL-17 production by immune cells. Our study identifies a novel signaling pathway, in which 20(OH)D3 and 20,23(OH)2D3 act as antagonists or inverse agonists of RORa and RORy, that opens new possibilities for local (skin) or systemic regulation. {\circledC} FASEB.",
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Słomiński, AT, Kim, T, Takeda, Y, Janjetovic, Z, Brozyna, AA, Skobowiat, C, Wang, J, Postlethwaite, AE, Li, W, Tuckey, R & Jetten, AM 2014, 'RORo: And ROR 7 are expressed in human skin and serve as receptors for endogenously produced noncalcemic 20-hydroxy- and 20,23-dihydroxyvitamin D' FASEB Journal, vol. 28, no. 7, pp. 2775-2789. https://doi.org/10.1096/fj.13-242040

RORo: And ROR 7 are expressed in human skin and serve as receptors for endogenously produced noncalcemic 20-hydroxy- and 20,23-dihydroxyvitamin D. / Słomiński, A.T.; Kim, T.; Takeda, Y.; Janjetovic, Z.; Brozyna, A.A.; Skobowiat, C.; Wang, J.; Postlethwaite, A.E.; Li, W.; Tuckey, Robert; Jetten, A.M.

In: FASEB Journal, Vol. 28, No. 7, 2014, p. 2775-2789.

Research output: Contribution to journalArticle

TY - JOUR

T1 - RORo: And ROR 7 are expressed in human skin and serve as receptors for endogenously produced noncalcemic 20-hydroxy- and 20,23-dihydroxyvitamin D

AU - Słomiński, A.T.

AU - Kim, T.

AU - Takeda, Y.

AU - Janjetovic, Z.

AU - Brozyna, A.A.

AU - Skobowiat, C.

AU - Wang, J.

AU - Postlethwaite, A.E.

AU - Li, W.

AU - Tuckey, Robert

AU - Jetten, A.M.

PY - 2014

Y1 - 2014

N2 - RORa and RORy are expressed in human skin cells that produce the noncalcemic 20-hydroxyvi-tamin D3 [20(OH)D3] and 20,23-dihydroxyvitamin D3 [20,23(OH)2D3]. Chinese hamster ovary (CHO) cells stably expressing a Tet-on RORa or RORy expression vector and a ROR-responsive element (RORE)-LUC reporter, and a mammalian 2-hybrid model examining the interaction between the ligand binding domain (LBD) of RORa or RORy with an LBD-interacting LXXLL-peptide, were used to study ROR-antagonist activities. These assays revealed that 20(OH)D3 and 20,23(OH)2D3 function as antagonists of RORa and RORy. Moreover, 20(OH)D3 inhibited the activation of the promoter of the Bmal1 and G6pase genes, targets of RORa, and 20(OH)D3 and 20,23(OH)2D3 inhibited Il17 promoter activity in Jurkat cells overexpressing RORa or RORy. Molecular modeling using crystal structures of the LBDs of RORa and RORy revealed docking scores for 20(OH)D3, 20, 23(OH)2D3 and 1,25(OH)2D3 similar to those of the natural ligands, predicting good binding to the receptor. Notably, 20(OH)D3, 20,23(OH)2D3, and 1,25(OH)2D3 inhibited RORE-mediated activation of a reporter in keratinocytes and melanoma cells and inhibited IL-17 production by immune cells. Our study identifies a novel signaling pathway, in which 20(OH)D3 and 20,23(OH)2D3 act as antagonists or inverse agonists of RORa and RORy, that opens new possibilities for local (skin) or systemic regulation. © FASEB.

AB - RORa and RORy are expressed in human skin cells that produce the noncalcemic 20-hydroxyvi-tamin D3 [20(OH)D3] and 20,23-dihydroxyvitamin D3 [20,23(OH)2D3]. Chinese hamster ovary (CHO) cells stably expressing a Tet-on RORa or RORy expression vector and a ROR-responsive element (RORE)-LUC reporter, and a mammalian 2-hybrid model examining the interaction between the ligand binding domain (LBD) of RORa or RORy with an LBD-interacting LXXLL-peptide, were used to study ROR-antagonist activities. These assays revealed that 20(OH)D3 and 20,23(OH)2D3 function as antagonists of RORa and RORy. Moreover, 20(OH)D3 inhibited the activation of the promoter of the Bmal1 and G6pase genes, targets of RORa, and 20(OH)D3 and 20,23(OH)2D3 inhibited Il17 promoter activity in Jurkat cells overexpressing RORa or RORy. Molecular modeling using crystal structures of the LBDs of RORa and RORy revealed docking scores for 20(OH)D3, 20, 23(OH)2D3 and 1,25(OH)2D3 similar to those of the natural ligands, predicting good binding to the receptor. Notably, 20(OH)D3, 20,23(OH)2D3, and 1,25(OH)2D3 inhibited RORE-mediated activation of a reporter in keratinocytes and melanoma cells and inhibited IL-17 production by immune cells. Our study identifies a novel signaling pathway, in which 20(OH)D3 and 20,23(OH)2D3 act as antagonists or inverse agonists of RORa and RORy, that opens new possibilities for local (skin) or systemic regulation. © FASEB.

U2 - 10.1096/fj.13-242040

DO - 10.1096/fj.13-242040

M3 - Article

VL - 28

SP - 2775

EP - 2789

JO - FASEB Journal

JF - FASEB Journal

SN - 0892-6638

IS - 7

ER -