Cited by 100Vitamin D Research StudiesPharmacogenetics and Drug MetabolismCholesterol and Lipid Metabolism

In vivo production of novel vitamin D2 hydroxy-derivatives by human placentas, epidermal keratinocytes Caco-2 colon cells and the adrenal gland

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Slominski, Andrzej T., Kim, Tae-Kang, Shehabi, Haleem Z., Tang, Edith K.Y., Benson, Heather A.E., Semak, Igor, Lin, Zongtao, Yates, Charles R. +3 more·2014·Molecular and Cellular Endocrinology

Abstract

We investigated the metabolism of vitamin D2 to hydroxyvitamin D2 metabolites ((OH)D2) by human placentas ex-utero, adrenal glands ex-vivo and cultured human epidermal keratinocytes and colonic Caco-2 cells, and identified 20(OH)D2, 17,20(OH)₂D2, 1,20(OH)₂D2, 25(OH)D2 and 1,25(OH)₂D2 as products. Inhibition of product formation by 22R-hydroxycholesterol indicated involvement of CYP11A1 in 20- and 17-hydroxylation of vitamin D2, while use of ketoconazole indicated involvement of CYP27B1 in 1α-hydroxylation of products. Studies with purified human CYP11A1 confirmed the ability of this enzyme to convert vitamin D2 to 20(OH)D2 and 17,20(OH)₂D2. In placentas and Caco-2 cells, production of 20(OH)D2 was higher than 25(OH)D2 while in human keratinocytes the production of 20(OH)D2 and 25(OH)D2 were comparable. HaCaT keratinocytes showed high accumulation of 1,20(OH)₂D2 relative to 20(OH)D2 indicating substantial CYP27B1 activity. This is the first in vivo evidence for a novel pathway of vitamin D2 metabolism initiated by CYP11A1 and modified by CYP27B1, with the product profile showing tissue- and cell-type specificity.

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Crossref · 10.1016/j.mce.2013.12.012

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  • Molecular modeling using the crystal structure of CYP11A1 predicts that other intermediates of cholesterol synthesis could also serve as substrates for CYP11A1.

    “Molecular modeling using the crystal structure of CYP11A1 predicts that other intermediates of cholesterol synthesis could also serve as substrates for CYP11A1.”
  • Recombinant human CYP11A1 shows a similar sequence of reactions for vitamin D2, with 20(OH)D2 as the major product and lower production of 17,20(OH)2D2 and 17,20,24(OH)3D2 and another unidentified D2 metabolite.

    “A similar sequence of reactions was seen for recombinant human CYP11A1 [9]. The major product was 20(OH)D2 with lower production of 17,20(OH)2D2 and 17,20,24 (OH)3D2 and another unidentified D2 metabolite [28,29].”
  • Recombinant human CYP11A1 hydroxylates the prodrug 1α(OH)D2 to 1,20(OH)2D2.

    “In addition, the prodrug, 1α (OH)D2, could be hydroxylated by recombinant human CYP11A1 to 1,20(OH)2D2 [9].”
  • Caco-2 cells produce more 20(OH)D2 and 17,20(OH)2D2 than HaCaT keratinocytes.

    “Also, production of 20(OH)D2 and 17,20(OH)2D2 was higher in Caco-2 cells than in HaCaT keratinocytes [9]”
  • HaCaT keratinocytes produce more 1,20(OH)2D2 than Caco-2 cells.

    “but higher production of 1,20(OH)2D2 was seen in HaCaT keratinocytes [9].”

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Research works printed but not linked: 2

  1. Holick MF Holick SA Tavela T Gallagher B Schnoes HK DeLuca HF 1975 Synthesis of (6–3H)-1alpha-hydroxyvitamin D3 and its metabolism in vivo to (3H)-1alpha,25-dihydroxyvitamin D3 Science 190 576 8 1188356
  2. Slominski A Janjetovic Z Tuckey RC Nguyen MN Bhattacharya KG Wang J Li W Jiao Y Gu W Brown M Postlethwaite AE 2013a 20S-Hydroxyvitamin D3, noncalcemic product of CYP11A1 action on vitamin D3, exhibits potent antifibrogenic activity in vivo J Clin Endocrinol Metab 98 E298 303 23295467 PMC3565109

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Authors
Slominski, A. T., Kim, T.-K., Shehabi, H. Z., Tang, E. K. Y., Benson, H. A. E., Semak, I., Lin, Z., Yates, C. R., Wang, J., Li, W., & Tuckey, R. C.
Affiliations
  • University of Tennessee Health Science Center (US)
  • Curtin University (AU)
  • The University of Western Australia (AU)
  • Belarusian State University (BY)
Corresponding author
  • Slominski, Andrzej T.
  • Tuckey, Robert C.
Venue
Molecular and Cellular Endocrinology
Publisher
Elsevier BV
Published
2014-03
Volume
383
Issue
1-2
Pages
181-192
ISSN
0303-7207 (ISSN-L)
OpenAlex
W1968274590
Cited by
100
References
54
Citations by year
2026: 1 · 2025: 3 · 2024: 4 · 2023: 9 · 2022: 10 · 2021: 15 · 2020: 14 · 2019: 6 · 2018: 2 · 2017: 7 · 2016: 11 · 2015: 8 · 2014: 10
FWCI
8.91
Type
journal article
Language
en
Open access
Locations
  • Molecular and Cellular Endocrinology (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
  • UWA Profiles and Research Repository (UWA) (submittedVersion)
  • eSpace (Curtin University) (submittedVersion)
Funders
  • National Institutes of Health (1s10od010678-01)
  • National Institute of Arthritis and Musculoskeletal and Skin Diseases (R01AR052190, R21AR063242, R01AR056666)
Topics
  • Health Sciences › Medicine › Pathology and Forensic Medicine › Vitamin D Research Studies
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Pharmacogenetics and Drug Metabolism
  • Health Sciences › Medicine › Surgery › Cholesterol and Lipid Metabolism
Keywords
25-hydroxyvitamin D2, CYP11A1, CYP27B1, 22R-hydroxycholesterol, ketoconazole, placenta, epidermal keratinocytes, Caco-2 cells, adrenal gland
MeSH terms
Epidermal Cells, Adrenal Glands, Animals, Cholesterol Side-Chain Cleavage Enzyme, Epidermis, Ergocalciferols, Female, Gene Expression Regulation, Humans, Hydroxycholesterols, Hydroxylation, Ketoconazole, Mitochondria, Organ Specificity, Placenta, Pregnancy, Recombinant Proteins, 25-Hydroxyvitamin D3 1-alpha-Hydroxylase, Signal Transduction, Keratinocytes, Rats, Wistar, Caco-2 Cells, Tissue Culture Techniques, Rats

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