Rapid genomic changes induced by PTH, SIK-inhibition, FGF23, and 1,25(OH)2D3 in the mouse kidney

In vivo ChIP-seq in mouse kidney: we examine the genomic recruitment of VDR, RXR, pCREB, CBP, CRTC2 (TORC2), SRC1, SRC3, BRD4, RNApolII, H3K9ac, H3K27ac, and H3K36me3 after intraperitoneal injection of 10 mg/kg body weight (bw) 1,25(OH)2D3 (in propylene glycol), 230 mg/kg bw PTH (1–84) (in phosphate buffered saline (PBS)), 50 mg/kg bw FGF23 (in PBS + 0.1% BSA), 30 mg/kg YKL-05-099 (PBS + 25mM HCl), 40 mg/kg SK-124 (15% HPBCD (hydroxypropyl β-cyclodextrin)), or vehicle (EtOH, PBS, or HPBCD). YKL-05-099 and SK-124 are salt-inducible kinase inhibitors that liberate CRTC2 from sequestration in the cytoplasm and allow CRTC2 translocation to the nucleus. Our findings show that these TFs and coactivators contribute to transcription of genes in the kidney involved in vitamin D metabolism, demonstrates SIK-inhibition as a key modulator of vitamin D metabolism, and provides molecular insight into the coordinated mechanistic actions of PTH, FGF23, and 1,25(OH)2D3 in the mouse kidney that regulate mineral homeostasis. All samples included were analyzed in biological triplicate (3 individual mice). ChIP assay was performed on mouse kidneys (2 per mouse) after intraperitoneal injection of 10 mg/kg body weight (bw) 1,25(OH)2D3 (in propylene glycol), 230 mg/kg bw PTH (1–84) (in phosphate buffered saline (PBS)), 50 mg/kg bw FGF23 (in PBS + 0.1% BSA), 30 mg/kg YKL-05-099 (PBS + 25mM HCl), 40 mg/kg SK-124 (15% HPBCD (hydroxypropyl β-cyclodextrin)), or vehicle (EtOH, PBS, or HPBCD). All treatements were 1 h unless otherwise indicated in treatment column. ChIP-seq libraries were then prepared for all samples and sequenced paired-end on the Illumina NovaSeq instrument. Input files, pCREB, histones with PTH treatment at 60 min included in GSE133025.