1β,25-Dihydroxyvitamin D3: A new vitamin D metabolite in human serum

Steven Pauwels, Ivo Jans, Jaak Billen, Annemieke Heijboer, Annemieke Verstuyf, Geert Carmeliet, Chantal Mathieu, Miguel Maestro, Etienne Waelkens, Pieter Evenepoel, Roger Bouillon, Dirk Vanderschueren, Pieter Vermeersch

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Background The measurement of 1α,25(OH)2D3 in human serum poses a true challenge as concentrations are very low and structurally similar metabolites can interfere. Materials and methods During optimization of our in-house LC–MSMS method for serum 1α,25(OH)2D3 a previously co-eluting isobaric interference was separated. The isobar was identified as 1β,25(OH)2D3 by comparing retention time and fragmentation spectra to standards (other isobaric dihydroxylated vitamin D3 analogs). 1β,25(OH)2D3 showed specific cluster formation (water), not present in 1α,25(OH)2D3. 1β,25(OH)2D3 was measured in serum of apparently healthy human volunteers (n = 20), patients with high serum 25-hydroxyvitamin D [25(OH)D] concentrations (>50 ng/mL) (n = 33 among which 4 with very high levels (>150 ng/mL)) and patients with kidney failure (n = 68; 39 stage 1–3, 29 stage 4–5). Pearson's r was calculated for correlations and Mann-Whitney statistic to compare group medians. Results Median serum 1β,25(OH)2D3 was 11 pg/mL in apparently healthy volunteers and increased to 20 pg/mL for serum 25(OH)D concentrations above 80 ng/mL (n = 22) (p < 0.0001). 1β,25(OH)2D3 concentrations were significantly correlated to serum 25(OH)D concentrations (r = 0.85) for the combined results from healthy volunteers and patient sera (n = 53) (p < 0.0001). For patients with kidney failure, median serum 1β,25(OH)2D3 was 7 pg/mL and not different from the median level in healthy volunteers (p = 0.06). The median concentration did not vary with different stages. Conclusions We present evidence for the widespread presence of 1β,25(OH)2D3, a new vitamin D metabolite, in human serum. The level increases with rising serum 25(OH)D concentrations and is particularly high in patients with very high 25(OH)D levels. We previously demonstrated that 1β,25(OH)2D3 is a poor genomic agonist but a potent non-genomic antagonist of 1α,25(OH)2D3. The clinical implications of the presence of this analog therefore require further exploration.

Original languageEnglish
Pages (from-to)341-348
Number of pages8
JournalJournal of Steroid Biochemistry and Molecular Biology
Volume173
DOIs
Publication statusPublished - 1 Oct 2017

Cite this

Pauwels, Steven ; Jans, Ivo ; Billen, Jaak ; Heijboer, Annemieke ; Verstuyf, Annemieke ; Carmeliet, Geert ; Mathieu, Chantal ; Maestro, Miguel ; Waelkens, Etienne ; Evenepoel, Pieter ; Bouillon, Roger ; Vanderschueren, Dirk ; Vermeersch, Pieter. / 1β,25-Dihydroxyvitamin D3 : A new vitamin D metabolite in human serum. In: Journal of Steroid Biochemistry and Molecular Biology. 2017 ; Vol. 173. pp. 341-348.
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title = "1β,25-Dihydroxyvitamin D3: A new vitamin D metabolite in human serum",
abstract = "Background The measurement of 1α,25(OH)2D3 in human serum poses a true challenge as concentrations are very low and structurally similar metabolites can interfere. Materials and methods During optimization of our in-house LC–MSMS method for serum 1α,25(OH)2D3 a previously co-eluting isobaric interference was separated. The isobar was identified as 1β,25(OH)2D3 by comparing retention time and fragmentation spectra to standards (other isobaric dihydroxylated vitamin D3 analogs). 1β,25(OH)2D3 showed specific cluster formation (water), not present in 1α,25(OH)2D3. 1β,25(OH)2D3 was measured in serum of apparently healthy human volunteers (n = 20), patients with high serum 25-hydroxyvitamin D [25(OH)D] concentrations (>50 ng/mL) (n = 33 among which 4 with very high levels (>150 ng/mL)) and patients with kidney failure (n = 68; 39 stage 1–3, 29 stage 4–5). Pearson's r was calculated for correlations and Mann-Whitney statistic to compare group medians. Results Median serum 1β,25(OH)2D3 was 11 pg/mL in apparently healthy volunteers and increased to 20 pg/mL for serum 25(OH)D concentrations above 80 ng/mL (n = 22) (p < 0.0001). 1β,25(OH)2D3 concentrations were significantly correlated to serum 25(OH)D concentrations (r = 0.85) for the combined results from healthy volunteers and patient sera (n = 53) (p < 0.0001). For patients with kidney failure, median serum 1β,25(OH)2D3 was 7 pg/mL and not different from the median level in healthy volunteers (p = 0.06). The median concentration did not vary with different stages. Conclusions We present evidence for the widespread presence of 1β,25(OH)2D3, a new vitamin D metabolite, in human serum. The level increases with rising serum 25(OH)D concentrations and is particularly high in patients with very high 25(OH)D levels. We previously demonstrated that 1β,25(OH)2D3 is a poor genomic agonist but a potent non-genomic antagonist of 1α,25(OH)2D3. The clinical implications of the presence of this analog therefore require further exploration.",
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author = "Steven Pauwels and Ivo Jans and Jaak Billen and Annemieke Heijboer and Annemieke Verstuyf and Geert Carmeliet and Chantal Mathieu and Miguel Maestro and Etienne Waelkens and Pieter Evenepoel and Roger Bouillon and Dirk Vanderschueren and Pieter Vermeersch",
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Pauwels, S, Jans, I, Billen, J, Heijboer, A, Verstuyf, A, Carmeliet, G, Mathieu, C, Maestro, M, Waelkens, E, Evenepoel, P, Bouillon, R, Vanderschueren, D & Vermeersch, P 2017, '1β,25-Dihydroxyvitamin D3: A new vitamin D metabolite in human serum' Journal of Steroid Biochemistry and Molecular Biology, vol. 173, pp. 341-348. https://doi.org/10.1016/j.jsbmb.2017.02.004

1β,25-Dihydroxyvitamin D3 : A new vitamin D metabolite in human serum. / Pauwels, Steven; Jans, Ivo; Billen, Jaak; Heijboer, Annemieke; Verstuyf, Annemieke; Carmeliet, Geert; Mathieu, Chantal; Maestro, Miguel; Waelkens, Etienne; Evenepoel, Pieter; Bouillon, Roger; Vanderschueren, Dirk; Vermeersch, Pieter.

In: Journal of Steroid Biochemistry and Molecular Biology, Vol. 173, 01.10.2017, p. 341-348.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - 1β,25-Dihydroxyvitamin D3

T2 - A new vitamin D metabolite in human serum

AU - Pauwels, Steven

AU - Jans, Ivo

AU - Billen, Jaak

AU - Heijboer, Annemieke

AU - Verstuyf, Annemieke

AU - Carmeliet, Geert

AU - Mathieu, Chantal

AU - Maestro, Miguel

AU - Waelkens, Etienne

AU - Evenepoel, Pieter

AU - Bouillon, Roger

AU - Vanderschueren, Dirk

AU - Vermeersch, Pieter

PY - 2017/10/1

Y1 - 2017/10/1

N2 - Background The measurement of 1α,25(OH)2D3 in human serum poses a true challenge as concentrations are very low and structurally similar metabolites can interfere. Materials and methods During optimization of our in-house LC–MSMS method for serum 1α,25(OH)2D3 a previously co-eluting isobaric interference was separated. The isobar was identified as 1β,25(OH)2D3 by comparing retention time and fragmentation spectra to standards (other isobaric dihydroxylated vitamin D3 analogs). 1β,25(OH)2D3 showed specific cluster formation (water), not present in 1α,25(OH)2D3. 1β,25(OH)2D3 was measured in serum of apparently healthy human volunteers (n = 20), patients with high serum 25-hydroxyvitamin D [25(OH)D] concentrations (>50 ng/mL) (n = 33 among which 4 with very high levels (>150 ng/mL)) and patients with kidney failure (n = 68; 39 stage 1–3, 29 stage 4–5). Pearson's r was calculated for correlations and Mann-Whitney statistic to compare group medians. Results Median serum 1β,25(OH)2D3 was 11 pg/mL in apparently healthy volunteers and increased to 20 pg/mL for serum 25(OH)D concentrations above 80 ng/mL (n = 22) (p < 0.0001). 1β,25(OH)2D3 concentrations were significantly correlated to serum 25(OH)D concentrations (r = 0.85) for the combined results from healthy volunteers and patient sera (n = 53) (p < 0.0001). For patients with kidney failure, median serum 1β,25(OH)2D3 was 7 pg/mL and not different from the median level in healthy volunteers (p = 0.06). The median concentration did not vary with different stages. Conclusions We present evidence for the widespread presence of 1β,25(OH)2D3, a new vitamin D metabolite, in human serum. The level increases with rising serum 25(OH)D concentrations and is particularly high in patients with very high 25(OH)D levels. We previously demonstrated that 1β,25(OH)2D3 is a poor genomic agonist but a potent non-genomic antagonist of 1α,25(OH)2D3. The clinical implications of the presence of this analog therefore require further exploration.

AB - Background The measurement of 1α,25(OH)2D3 in human serum poses a true challenge as concentrations are very low and structurally similar metabolites can interfere. Materials and methods During optimization of our in-house LC–MSMS method for serum 1α,25(OH)2D3 a previously co-eluting isobaric interference was separated. The isobar was identified as 1β,25(OH)2D3 by comparing retention time and fragmentation spectra to standards (other isobaric dihydroxylated vitamin D3 analogs). 1β,25(OH)2D3 showed specific cluster formation (water), not present in 1α,25(OH)2D3. 1β,25(OH)2D3 was measured in serum of apparently healthy human volunteers (n = 20), patients with high serum 25-hydroxyvitamin D [25(OH)D] concentrations (>50 ng/mL) (n = 33 among which 4 with very high levels (>150 ng/mL)) and patients with kidney failure (n = 68; 39 stage 1–3, 29 stage 4–5). Pearson's r was calculated for correlations and Mann-Whitney statistic to compare group medians. Results Median serum 1β,25(OH)2D3 was 11 pg/mL in apparently healthy volunteers and increased to 20 pg/mL for serum 25(OH)D concentrations above 80 ng/mL (n = 22) (p < 0.0001). 1β,25(OH)2D3 concentrations were significantly correlated to serum 25(OH)D concentrations (r = 0.85) for the combined results from healthy volunteers and patient sera (n = 53) (p < 0.0001). For patients with kidney failure, median serum 1β,25(OH)2D3 was 7 pg/mL and not different from the median level in healthy volunteers (p = 0.06). The median concentration did not vary with different stages. Conclusions We present evidence for the widespread presence of 1β,25(OH)2D3, a new vitamin D metabolite, in human serum. The level increases with rising serum 25(OH)D concentrations and is particularly high in patients with very high 25(OH)D levels. We previously demonstrated that 1β,25(OH)2D3 is a poor genomic agonist but a potent non-genomic antagonist of 1α,25(OH)2D3. The clinical implications of the presence of this analog therefore require further exploration.

KW - 1,25-dihydroxyvitamin D

KW - Liquid chromatography

KW - Mass Spectrometry

KW - Metabolism

KW - Vitamin D

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