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. 2021 Sep 4;22(17):9603.
doi: 10.3390/ijms22179603.

High-Throughput Gene and Protein Analysis Revealed the Response of Disc Cells to Vitamin D, Depending on the VDR FokI Variants

Alessandra Colombini  1 , Paola De Luca  1 , Davide Cangelosi  2 , Carlotta Perucca Orfei  1 , Enrico Ragni  1 , Marco Viganò  1 , Michela Malacarne  2 , Mauro Castagnetta  3 , Marco Brayda-Bruno  4 , Domenico Coviello  2 , Laura de Girolamo  1
Affiliations

High-Throughput Gene and Protein Analysis Revealed the Response of Disc Cells to Vitamin D, Depending on the VDR FokI Variants

Alessandra Colombini et al. Int J Mol Sci. .

Abstract

Vitamin D showed a protective effect on intervertebral disc degeneration (IDD) although conflicting evidence is reported. An explanation could be due to the presence of the FokI functional variant in the vitamin D receptor (VDR), observed as associated with spine pathologies. The present study was aimed at investigating-through high-throughput gene and protein analysis-the response of human disc cells to vitamin D, depending on the VDR FokI variants. The presence of FokI VDR polymorphism was determined in disc cells from patients with discopathy. 1,25(OH)2D3 was administered to the cells with or without interleukin 1 beta (IL-1β). Microarray, protein arrays, and multiplex protein analysis were performed. In both FokI genotypes (FF and Ff), vitamin D upregulated metabolic genes of collagen. In FF cells, the hormone promoted the matrix proteins synthesis and a downregulation of enzymes involved in matrix catabolism, whereas Ff cells behaved oppositely. In FF cells, inflammation seems to hamper the synthetic activity mediated by vitamin D. Angiogenic markers were upregulated in FF cells, along with hypertrophic markers, some of them upregulated also in Ff cells after vitamin D treatment. Higher inflammatory protein modulation after vitamin D treatment was observed in inflammatory condition. These findings would help to clarify the clinical potential of vitamin D supplementation in patients affected by IDD.

Keywords: gene profile; inflammation; intervertebral disc; protein profile; vitamin D; vitamin D receptor gene polymorphism.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Upregulated genes and related pathways in homozygous FF disc cells. Pie chart obtained by Panther.
Figure 2
Figure 2
Downregulated genes and related pathways in heterozygous Ff disc cells. Pie chart obtained by Panther.
Figure 3
Figure 3
Upregulated genes and related pathways in heterozygous Ff disc cells. Pie chart obtained by Panther.
Figure 4
Figure 4
Downregulated genes and related pathways in homozygous FF disc cells. Pie chart obtained by Panther.
Figure 5
Figure 5
Pathways associated to the genes modulated by inflammatory stimulus in homozygous FF disc cells. Relevant pathways are highlighted in black bold. Pie chart obtained by Panther.
Figure 6
Figure 6
Pathways associated to the genes modulated by inflammatory stimulus in heterozygous Ff disc cells. Relevant pathways are highlighted in black bold. Pie chart obtained by Panther.
Figure 7
Figure 7
Heat maps shows relevant group of genes differently modulated by vitamin D depending on FokI genotype in basal and inflamed conditions. Heat maps obtained by Morpheus, https://software.broadinstitute.org/morpheus (Date of access 6 April 2021).
Figure 8
Figure 8
Levels of matrix-metalloproteases and BMP-2 secreted by pooled, NP, AF, and EP cells of 7 donors in basal condition. Arrows indicate significant differences and tendencies in protein secretion of vitamin D treated cells in comparison with untreated cells. * p ≤ 0.05, ** p ≤ 0.01.
Figure 9
Figure 9
Levels of specific proteins secreted by pooled, NP, AF, and EP cells of seven donors in inflamed condition. Arrows indicate significant differences and tendencies in protein secretion of vitamin D treated cells in comparison with untreated cells. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.
See this image and copyright information in PMC

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