10.5061/DRYAD.41NS1RNBP
Janjetovic, Zorica
University of Alabama at Birmingham
Postlethwaite, Arnold Postlethwaite
0000-0002-5952-0161
University of Tennessee Health Science Center
Kang, Hong Soon
0000-0003-2230-8797
National Institutes of Health
Kim, Tae Kang
0000-0003-1394-4134
University of Alabama at Birmingham
Tuckey, Robert
0000-0002-5022-4729
University of Western Australia
Crossman, David
University of Alabama at Birmingham
Jetten, Anton
National Institutes of Health
Alominski, Andrzej
0000-0001-8963-3995
University of Alabama at Birmingham
Supplemental tables from: Antifibrogenic activities of CYP11A1-derived
vitamin D3-hydroxyderivatives are dependent on RORγ
Dryad
dataset
2020
National Cancer Institute
https://ror.org/040gcmg81
1RO1 AR052190 and 1R21 AR0665051
United States Department of Veterans Affairs
https://ror.org/05rsv9s98
1I01BX004293-01A1
National Cancer Institute
https://ror.org/040gcmg81
1R01AR073004-01A1 and R01AR071189-01A1
2020-08-18T00:00:00Z
2020-08-18T00:00:00Z
en
102404 bytes
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CC0 1.0 Universal (CC0 1.0) Public Domain Dedication
Previous studies showed that non-calcemic 20(OH)D3, a product of CYP11A1
action on vitamin D3, has antifibrotic activity in human dermal
fibroblasts and in a bleomycin mouse model of scleroderma. In this study
we tested the role of RORγ, which is expressed in skin, in the action of
CYP11A1-derived secosteroids using murine fibroblasts isolated from the
skin of wild type (RORg+/+), knock out (RORg-/-) and heterozygote
(RORg+/-) mice. CYP11A1-derived 20(OH)D3, 20,23(OH)2D3, 1,20(OH)2D3, and
1,20,23(OH)3D3 inhibited proliferation of RORγ+/+ fibroblasts in a
dose-dependent manner with a similar potency to 1,25(OH)2D3. Surprisingly,
this effect was reversed in RORγ+/- and RORγ-/- fibroblasts with the most
pronounced stimulatory effect seen in RORγ-/- fibroblasts. All of the
analogs tested inhibited TGF-β1-induced collagen synthesis in RORγ+/+
fibroblasts and the expression of other fibrosis-related genes. This
effect was curtailed or reversed in RORγ-/- fibroblasts. These results
show that the antiproliferative and antifibrotic activities of the vitamin
D hydroxy-derivatives are dependent on a functional RORγ. The dramatic
changes in the transcriptomes of fibroblasts of RORg-/- versus wild type
mice following treatment with 20(OH)D3 or 1,20(OH)2D3 provide a molecular
basis to explain, at least in part, the observed phenotypic differences
Data were analyzed with Ingenuity Pathway Analysis (Ingenuity® Systems,
www.ingenuity.com). For generating networks, a data set containing gene
identifiers and corresponding expression values was uploaded into the
application. Each identifier was mapped to its corresponding object in
Ingenuity’s Knowledge Base. A fold change cutoff of +/-2 was set to
identify molecules whose expression was significantly differentially
regulated. These molecules, called Network Eligible molecules, were
overlaid onto a global molecular network developed from information
contained in Ingenuity’s Knowledge Base. Networks of Network Eligible
Molecules were then algorithmically generated based on their connectivity.
The Functional Analysis identified the biological functions and/or
diseases that were most significant to the entire data set. Molecules from
the dataset that met the fold change cutoff of +/-2 and were associated
with biological functions and/or diseases in Ingenuity’s Knowledge Base
were considered for the analysis. Right-tailed Fisher’s exact test was
used to calculate the p-value determining the probability that each
biological function and/or disease assigned to that data set is due to
chance alone.