Vitamin D receptor cross-talk with p63 signaling promotes epidermal cell fate - PubMed (original) (raw)

Vitamin D receptor cross-talk with p63 signaling promotes epidermal cell fate

Yuko Oda et al. J Steroid Biochem Mol Biol. 2023 Sep.

Abstract

The vitamin D receptor with its ligand 1,25 dihydroxy vitamin D3 (1,25D3) regulates epidermal stem cell fate, such that VDR removal from Krt14 expressing keratinocytes delays re-epithelialization of epidermis after wound injury in mice. In this study we deleted Vdr from Lrig1 expressing stem cells in the isthmus of the hair follicle then used lineage tracing to evaluate the impact on re-epithelialization following injury. We showed that Vdr deletion from these cells prevents their migration to and regeneration of the interfollicular epidermis without impairing their ability to repopulate the sebaceous gland. To pursue the molecular basis for these effects of VDR, we performed genome wide transcriptional analysis of keratinocytes from Vdr cKO and control littermate mice. Ingenuity Pathway analysis (IPA) pointed us to the TP53 family including p63 as a partner with VDR, a transcriptional factor that is essential for proliferation and differentiation of epidermal keratinocytes. Epigenetic studies on epidermal keratinocytes derived from interfollicular epidermis showed that VDR is colocalized with p63 within the specific regulatory region of MED1 containing super-enhancers of epidermal fate driven transcription factor genes such as Fos and Jun. Gene ontology analysis further implicated that Vdr and p63 associated genomic regions regulate genes involving stem cell fate and epidermal differentiation. To demonstrate the functional interaction between VDR and p63, we evaluated the response to 1,25(OH)2D3 of keratinocytes lacking p63 and noted a reduction in epidermal cell fate determining transcription factors such as Fos, Jun. We conclude that VDR is required for the epidermal stem cell fate orientation towards interfollicular epidermis. We propose that this role of VDR involves cross-talk with the epidermal master regulator p63 through super-enhancer mediated epigenetic dynamics.

Keywords: Cell fate; Epigenetics; Keratinocytes, wounding; P63; Vitamin D; Vitamin D receptor.

Copyright © 2023. Published by Elsevier Ltd.

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Figures

Fig. 1.

Fig. 1.

Ablation of Vdr impairs iSC movement to interfollicular epidermis but not to SG during wound re-epithelialization. a Representative images of 2d (upper panels) and 30 or 35 days after skin wounding (lower panel). TdT labeled iSCs after 2 d migrated to the IFE in CON (white triangles, upper panels) but less so in cKO (bottom panels). TdT iSC regenerate SG similarly in CON and cKO. (b) Schematic localization of Lrig1 expressing isthmus stem cells, SG, IFE epidermis in skin. c Quantification of the TdT labeled cells in the IFE epidermis and SG was determined in 8–10 different fields for control and cKO mice (2 each). Statistical significance was calculated by t-test (* P < 0.05).

Fig. 2.

Fig. 2.

a Venn diagram shows the number of VDR and p63 peaks in the absence (vehicle control, left panel) and presence of 1,25D3 (right) in primary keratinocytes. The increased number of 799 VDR/p63 overlap sites is shown by a pink box. b Gene Ontology (GO) analysis performed on the annotated set of genes from the 799 VDR/p63 overlapped peaks. Top biological processes related to skin wound healing are pink-highlighted, in which GO terms and p-values are shown. The associated genes are listed in Supplemental Table S1.

Fig. 3.

Fig. 3.

a ChIP-seq profiles for keratinocytes treated with vehicle (yellow) or 1,25D3 (blue) and their overlap are shown by green. The VDR, p63, CTCF, and Med1 peaks for the FOS gene, in which the colocalization of VDR/p63 peaks was marked by yellow highlights, and location of Med1 enriched super-enhancers identified by Rose software was marked by a red bar. b The locations of binding motifs for VDR, p63 and TCF4 are shown at – 24 kb upstream region of the FOS gene locus.

Fig. 4.

Fig. 4.

Decreased p63 expression affects the 1,25D3 response in fate transcription factors of FOS and JUN in keratinocytes. a Decreased protein expression of p63 in the shp63 cell line compared to the parent line TERT, in which actin was used as a control. b. mRNA expression of FOS, JUN, SOX7 and CAMP in shp63 and TERT keratinocytes treated with vehicle and 1 × 10−8 M 1,25D3 overnight. Gene expression is evaluated by qPCR, and fold induction compared to control TERT cells was calculated. Averages of three batches of 1,25D3 induction were calculated t-test * <0.05.

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