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Vitamin D metabolites inhibit hepatitis C virus and modulate cellular gene expression

Vitamin D metabolites inhibit hepatitis C virus and modulate cellular gene expression
Vitamin D metabolites inhibit hepatitis C virus and modulate cellular gene expression
Background and Aims: Previous studies suggest that low serum 25-hydroxyvitamin D [25(OH) D] levels are associated with reduced responsiveness to interferon and ribavirin therapy. We investigated the impact of vitamin D metabolites on HCV and cellular gene expression in cultured hepatoma cells.

Methods: HCV Replicon cell lines stably expressing luciferase reporter constructs (genotype 1b and 2a replicon) or JC1-Luc2a were incubated in the presence of vitamin D2, vitamin D3 or 1,25-dihydroxyvitamin D3 (1,25(OH)2D3). Presence of HCV was quantified by a luciferase reporter assay and immunoblot of the Core protein. Synergy of interferon-alpha A/D (IFN-?) and 1,25(OH)2D3 was evaluated using the Chou-Talalay method. Cellular gene expression by microarray analysis using Illumina Bead Chips and real-time quantitative PCR.

Results: Vitamin D2, D3 and 1,25(OH)2D3 each demonstrated anti- HCV activity at low micro molar concentrations. In vitro conversion from D3 to 25(OH)D3 was shown by LC/MS/MS. Combination indices of 1,25(OH)2D3 and IFN-? demonstrated a synergistic effect (0.23-0.46) and significantly reduced core expression by immunoblot. Differentially expressed genes were identified between Huh7.5.1 cells in the presence and absence of 1,25(OH)2D3 and HCV. Genes involved with classical effects of vitamin D metabolism and excretion were activated, along with genes linked to autophagy such as G-protein coupled receptor 37 (GPR37) and Hypoxiainducible factor 1-alpha (HIF1a). Additionally, additive effects of 1,25(OH)2D3 and IFN-? were seen on mRNA expression of chemokine motif ligand 20 (CCL20).

Conclusions: This study shows that vitamin D reduces HCV protein production in cell culture synergistically with IFN-?. Vitamin D also activates gene expression independently and additively with IFN-? and this may explain its ability to aid in the clearance of HCV in vivo.
Gutierrez, J.A.
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Jones, K.A.
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Flores, R.
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Singhania, A.
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Woelk, C.H.
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Schooley, R.T.
a5fa4402-5e04-4f70-8a8d-92035524ff12
Wyles, D.L
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Gutierrez, J.A.
c8bea97c-504e-4baa-b9a1-73451f558165
Jones, K.A.
2333d02e-c72b-4420-9b36-a554915b8e3d
Flores, R.
580e477b-714c-45eb-93f8-f77bc3fe41ba
Singhania, A.
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Woelk, C.H.
4d3af0fd-658f-4626-b3b5-49a6192bcf7d
Schooley, R.T.
a5fa4402-5e04-4f70-8a8d-92035524ff12
Wyles, D.L
c777d43c-1f22-40e3-81e0-803b76a8583b

Gutierrez, J.A., Jones, K.A., Flores, R., Singhania, A., Woelk, C.H., Schooley, R.T. and Wyles, D.L (2014) Vitamin D metabolites inhibit hepatitis C virus and modulate cellular gene expression. Journal of Virology & Antiviral Research, 3 (3). (doi:10.4172/2324-8955.1000129).

Record type: Article

Abstract

Background and Aims: Previous studies suggest that low serum 25-hydroxyvitamin D [25(OH) D] levels are associated with reduced responsiveness to interferon and ribavirin therapy. We investigated the impact of vitamin D metabolites on HCV and cellular gene expression in cultured hepatoma cells.

Methods: HCV Replicon cell lines stably expressing luciferase reporter constructs (genotype 1b and 2a replicon) or JC1-Luc2a were incubated in the presence of vitamin D2, vitamin D3 or 1,25-dihydroxyvitamin D3 (1,25(OH)2D3). Presence of HCV was quantified by a luciferase reporter assay and immunoblot of the Core protein. Synergy of interferon-alpha A/D (IFN-?) and 1,25(OH)2D3 was evaluated using the Chou-Talalay method. Cellular gene expression by microarray analysis using Illumina Bead Chips and real-time quantitative PCR.

Results: Vitamin D2, D3 and 1,25(OH)2D3 each demonstrated anti- HCV activity at low micro molar concentrations. In vitro conversion from D3 to 25(OH)D3 was shown by LC/MS/MS. Combination indices of 1,25(OH)2D3 and IFN-? demonstrated a synergistic effect (0.23-0.46) and significantly reduced core expression by immunoblot. Differentially expressed genes were identified between Huh7.5.1 cells in the presence and absence of 1,25(OH)2D3 and HCV. Genes involved with classical effects of vitamin D metabolism and excretion were activated, along with genes linked to autophagy such as G-protein coupled receptor 37 (GPR37) and Hypoxiainducible factor 1-alpha (HIF1a). Additionally, additive effects of 1,25(OH)2D3 and IFN-? were seen on mRNA expression of chemokine motif ligand 20 (CCL20).

Conclusions: This study shows that vitamin D reduces HCV protein production in cell culture synergistically with IFN-?. Vitamin D also activates gene expression independently and additively with IFN-? and this may explain its ability to aid in the clearance of HCV in vivo.

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More information

Accepted/In Press date: 3 October 2014
e-pub ahead of print date: 2014
Published date: 6 October 2014
Organisations: Clinical & Experimental Sciences

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Local EPrints ID: 379212
URI: https://eprints.soton.ac.uk/id/eprint/379212
PURE UUID: 03b0c956-cc30-4271-b185-079b7b665c4a

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Date deposited: 18 Jul 2015 14:32
Last modified: 07 Jun 2018 16:30

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Contributors

Author: J.A. Gutierrez
Author: K.A. Jones
Author: R. Flores
Author: A. Singhania
Author: C.H. Woelk
Author: R.T. Schooley
Author: D.L Wyles

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