| Title: |
Vitamin D and its in vitro therapeutic action mediated through VDR rather than PDIA3 |
| Authors: |
Pyburn, Jaeden S; Hagg, T.; Keasey, M. P. |
| Source: |
Appalachian Student Research Forum [2018-2024] |
| Publisher Information: |
Digital Commons @ East Tennessee State University |
| Publication Year: |
2022 |
| Collection: |
Digital Commons @ East Tennessee State University |
| Subject Terms: |
Vitamin D; calcification; PDIA3; VDR; Cell Biology |
| Description: |
Brain calcification is a common occurrence in the aging process, with over 20% of individuals over the age of 65 showing hardened plaques in the basal ganglia. Loss of the vitamin D receptor (VDR) in transgenic mice leads to formation of calcified plaques in the basal ganglia and thalamus within the mice. Vitamin D signals through two known vitamin D responsive proteins, protein disulfide isomerase A3 (PDIA3) and VDR. In vitro, vitamin D has been demonstrated to suppress calcification in osteoblast-like cells. Here, we aim to elucidate which of PDIA3 or VDR transduce vitamin D mediated suppression of calcification in vitro. PDIA3 or VDR were selectively knocked out in human osteosarcoma (SaOs) cells using CRISPR/CAS9 technology to generate PDIA3 -/- or VDR -/- cells. Knockout for PDIA3 or VDR was confirmed by RT-qPCR assay or western blot analysis. The calcification of SaOs-2 cells was induced with treatment of β-glycerophosphate along with ascorbic acid allowing for determination of whether loss of PDIA3 or VDR would lead to altered calcium deposition. Cells null for PDIA3 but not VDR grew at a significantly slower rate than wild-type (WT) cells. Intriguingly, PDIA3 and VDR -/- cells displayed significantly more calcification relative to WT control cells. Calcitriol or the synthetic analogue EB-1089 suppressed calcification in vitro in WT and PDIA3 -/- but not VDR -/- cells as measured by alizarin red staining. These data suggest VDR is critical for mediating vitamin D’s inhibition of calcification in vitro, and that PDIA3 has a role in suppressing calcification. This study provides novel insights into vitamin D signaling and provides a foundation for further study and understanding of vitamin D related pathologies. |
| Document Type: |
text |
| Language: |
unknown |
| Availability: |
https://dc.etsu.edu/asrf/2022-boland/schedule/13 |
| Accession Number: |
edsbas.C7BA42F1 |
| Database: |
BASE |