In silico identification of novel transcription factors associates with CYP27B1 transcriptional regulation in mononuclear phagocytes exposed to LPS

MARTINELLI, R.; RODRIGUEZ, M.J.; DAURELIO, L.D.; ESTEBAN, L.

New York

Workshop; THE 22ND VITAMIN D WORKSHOP; 2019

Workshop Executive Committee (WEC)

Local production of 1,25-dihydroxyvitamin D (1,25(OH)(2)D), which is regulated by the CYP27B1 enzyme in mononuclear phagocytes, contributes to the immunomodulatory effects of vitamin D. The aim of this work was to characterize the transcriptional regulation of CYP27B1 gene in mononuclear phagocytes exposed to LPS, due to the relevance to identify the binding motifs of transcription factors to better understand the regulatory networks associated with inflammation,. First, the CYP27B1 human (AF500480) and mouse (AF041256) promoter sequences were analyzed using MatInspector and Pro-Coffee. Then, using microarray experiments were obtained from GEO, a meta-analysis was performed to obtain a global view of the gene expression before the stimulation of LPS in dendritic cells, monocytes, and macrophages. Finally, two experiments, GSE40885 and a time series (GSE19765), were analyzed in depth by differential expression analysis. Out of the two hundred putative TFBS found, an over expression of NFkB, CEBPB, and ATF3 in all the analyzes was observed, which also present putative TFBS and check according to the literature. However, consistently in all the analyzes, it was found over expression of two TF (PLAGL2 and STAT4) which present putative TFBS on the functional rSNPs (-1260 (rs10877012) and -925 (rs37822130). This data suggest that PLAGL2 and STAT4 are novel transcription factor which would participate in the transcriptional regulation of CYP27B1 in cells exposed to LPS. These FT, in turn, would be interacting with regions that present polymorphisms in the population and that could explain pathological phenotypes associated with the metabolism of vitamin D.