Transcriptome-Wide Profile of 25-Hydroxyvitamin D₃ in Primary Immune Cells from Human Peripheral Blood

Vitamin D₃ is an essential micronutrient mediating pleiotropic effects in multiple tissues and cell types via its metabolite 1α,25-dihydroxyvitamin D₃ (1,25(OH)₂D₃), which activates the transcription factor vitamin D receptor. In this study, we used peripheral blood mononuclear cells (PBMCs) obtained from five healthy adults and investigated transcriptome-wide, whether the precursor of 1,25(OH)₂D₃, 25-hydroxyvitamin D₃ (25(OH)D₃), has gene regulatory potential on its own. Applying thresholds of >2 in fold change of gene expression and <0.05 as a false discovery rate, in this ex vivo approach the maximal physiological concentration of 25(OH)D₃ (250 nM (nmol/L)) none of the study participants had a significant effect on their PBMC transcriptome. In contrast, 1000 and 10,000 nM 25(OH)D₃ regulated 398 and 477 genes, respectively, which is comparable to the 625 genes responding to 10 nM 1,25(OH)₂D₃. The majority of these genes displayed specificity to the tested individuals, but not to the vitamin D metabolite. Interestingly, the genes MYLIP (myosin regulatory light chain interacting protein) and ABCG1 (ATP binding cassette subfamily G member 1) showed to be specific targets of 10,000 nM 25(OH)D₃. In conclusion, 100- and 1000-fold higher 25(OH)D₃ concentrations than the reference 10 nM 1,25(OH)₂D₃ are able to affect the transcriptome of PBMCs with a profile comparable to that of 1,25(OH)₂D₃.