Vitamin D3 transactivates the zinc and manganese transporter SLC30A10 via the Vitamin D receptor.
Vitamin D3 regulates genes critical for human health and its deficiency is associated with an increased risk for osteoporosis, cancer, diabetes, multiple sclerosis, hypertension, inflammatory and immunological diseases. To study the impact of vitamin D3 on genes relevant for the transport and metabolism of nutrients and drugs, we employed next-generation sequencing (NGS) and analyzed global gene expression of the human-derived Caco-2 cell line treated with 500nM vitamin D3. Genes involved in neuropeptide signaling, inflammation, cell adhesion and morphogenesis were differentially expressed. Notably, genes implicated in zinc, manganese and iron homeostasis were largely increased by vitamin D3 treatment. An ∼10-fold increase in ceruloplasmin and ∼4-fold increase in haptoglobin gene expression suggested a possible association between vitamin D and iron homeostasis. SLC30A10, the gene encoding the zinc and manganese transporter ZnT10, was the chiefly affected transporter, with ∼15-fold increase in expression. SLC30A10 is critical for zinc and manganese homeostasis and mutations in this gene, resulting in impaired ZnT10 function or expression, cause manganese intoxication, with Parkinson-like symptoms. Our NGS results were validated by real-time PCR in Caco-2 cells, as well as in duodenal biopsies taken from healthy human subjects treated with 0.5μg vitamin D3 daily for 10 days. In addition to increasing gene expression of SLC30A10 and the positive control TRPV6, vitamin D3 also increased ZnT10 protein expression, as indicated by Western blot and cytofluorescence. In silico identification of potential vitamin D responsive elements (VDREs) in the 5'-flanking region of the SLC30A10 promoter and dual-luciferase reporter assay showed enhanced promoter activity in the presence of vitamin D receptor (VDR) and retinoid X receptor (RXR) constructs, as well as vitamin D3, but not when one of these factors was absent. Electrophoretic mobility shift assay (EMSA) and competition EMSA revealed binding of select sequences, namely, nt -1623/-1588 and nt -1758/-1723 relative to the transcription start site, to VDR-containing nuclear extracts. In conclusion, we have shown that vitamin D3 transactivates the SLC30A10 gene in a VDR-dependent manner, resulting in increased ZnT10 protein expression. Because SLC30A10 is highly expressed in the small intestine, it is possible that the control of zinc and manganese systemic levels is regulated by vitamin D3 in the intestine. Zinc, manganese and vitamin D are important for bone metabolism and brain health. Future examination of a possible role for supplementation or chelation of zinc and manganese, alongside vitamin D3 administration, will further our understanding of its potential benefit in the treatment of specific illnesses, such as osteoporosis and Parkinson's disease.
Department of Clinical Pharmacology and Toxicology, University Hospital Zurich, University of Zurich, Switzerland. Electronic address: taticlaro@yahoo.com.
Tatiana
T
Claro da Silva
Department of Clinical Pharmacology and Toxicology, University Hospital Zurich, University of Zurich, Switzerland. Electronic address: christian.hiller@usz.ch.
Christian
C
Hiller
Department of Clinical Pharmacology and Toxicology, University Hospital Zurich, University of Zurich, Switzerland. Electronic address: zhibo.gai@usz.ch.
Zhibo
Z
Gai
Department of Clinical Pharmacology and Toxicology, University Hospital Zurich, University of Zurich, Switzerland. Electronic address: gerd.kullak@usz.ch.
Gerd A
GA
Kullak-Ublick
D000328:Adult; D001665:Binding Sites; D018938:Caco-2 Cells; D002117:Calcitriol / Q000008:administration & dosage*; D015220:Calcium Channels / Q000235:genetics / Q000378:metabolism; D027682:Cation Transport Proteins / Q000819:agonists / Q000235:genetics* / Q000378:metabolism; D002570:Ceruloplasmin / Q000235:genetics / Q000378:metabolism; D004386:Duodenum / Q000187:drug effects* / Q000378:metabolism; D005260:Female; D063990:Gene Ontology; D017930:Genes, Reporter; D006242:Haptoglobins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007501:Iron / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008297:Male; D008345:Manganese / Q000378:metabolism; D058977:Molecular Sequence Annotation; D011446:Prospective Studies; D011485:Protein Binding; D018167:Receptors, Calcitriol / Q000819:agonists / Q000235:genetics* / Q000378:metabolism; D047488:Retinoid X Receptors / Q000235:genetics / Q000378:metabolism; D050916:TRPV Cation Channels / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation / Q000187:drug effects*; D015032:Zinc / Q000378:metabolism; D000075703:Zinc Transporter 8
D015220:Calcium Channels; D027682:Cation Transport Proteins; C485359:HP protein, human; D006242:Haptoglobins; D018167:Receptors, Calcitriol; D047488:Retinoid X Receptors; C492522:SLC30A8 protein, human; D050916:TRPV Cation Channels; C422827:TRPV6 protein, human; C579457:VDR protein, human; D000075703:Zinc Transporter 8; D008345:Manganese; D007501:Iron; D008156:Luciferases; D002570:Ceruloplasmin; D002117:Calcitriol; D015032:Zinc
Iron; Manganese; SLC30A10; VDRE; Vitamin D; Zinc