Hedgehog Signaling Inhibition by Smoothened Antagonist BMS-833923 Reduces Osteoblast Differentiation and Ectopic Bone Formation of Human Skeletal (Mesenchymal) Stem Cells.
Journal
Stem cells international
ISSN: 1687-966X
Titre abrégé: Stem Cells Int
Pays: United States
ID NLM: 101535822
Informations de publication
Date de publication:
2019
2019
Historique:
received:
24
06
2019
revised:
29
10
2019
accepted:
01
11
2019
entrez:
25
12
2019
pubmed:
25
12
2019
medline:
25
12
2019
Statut:
epublish
Résumé
Hedgehog (Hh) signaling is essential for osteoblast differentiation of mesenchymal progenitors during endochondral bone formation. However, the critical role of Hh signaling during adult bone remodeling remains to be elucidated. A Smoothened (SMO) antagonist/Hedgehog inhibitor, BMS-833923, identified during a functional screening of a stem cell signaling small molecule library, was investigated for its effects on the osteoblast differentiation of human skeletal (mesenchymal) stem cells (hMSC). Alkaline phosphatase (ALP) activity and Alizarin red staining were employed as markers for osteoblast differentiation and in vitro mineralization capacity, respectively. Global gene expression profiling was performed using the Agilent® microarray platform. Effects on in vivo ectopic bone formation were assessed by implanting hMSC mixed with hydroxyapatite-tricalcium phosphate granules subcutaneously in 8-week-old female nude mice, and the amount of bone formed was assessed using quantitative histology. BMS-833923, a SMO antagonist/Hedgehog inhibitor, exhibited significant inhibitory effects on osteoblast differentiation of hMSCs reflected by decreased ALP activity, in vitro mineralization, and downregulation of osteoblast-related gene expression. Similarly, we observed decreased in vivo ectopic bone formation. Global gene expression profiling of BMS-833923-treated compared to vehicle-treated control cells, identified 348 upregulated and 540 downregulated genes with significant effects on multiple signaling pathways, including GPCR, endochondral ossification, RANK-RANKL, insulin, TNF alpha, IL6, and inflammatory response. Further bioinformatic analysis employing Ingenuity Pathway Analysis revealed significant enrichment in BMS-833923-treated cells for a number of functional categories and networks involved in connective and skeletal tissue development and disorders, e.g., NF We identified SMO/Hedgehog antagonist (BMS-833923) as a powerful inhibitor of osteoblastic differentiation of hMSC that may be useful as a therapeutic option for treating conditions associated with high heterotopic bone formation and mineralization.
Sections du résumé
BACKGROUND
BACKGROUND
Hedgehog (Hh) signaling is essential for osteoblast differentiation of mesenchymal progenitors during endochondral bone formation. However, the critical role of Hh signaling during adult bone remodeling remains to be elucidated.
METHODS
METHODS
A Smoothened (SMO) antagonist/Hedgehog inhibitor, BMS-833923, identified during a functional screening of a stem cell signaling small molecule library, was investigated for its effects on the osteoblast differentiation of human skeletal (mesenchymal) stem cells (hMSC). Alkaline phosphatase (ALP) activity and Alizarin red staining were employed as markers for osteoblast differentiation and in vitro mineralization capacity, respectively. Global gene expression profiling was performed using the Agilent® microarray platform. Effects on in vivo ectopic bone formation were assessed by implanting hMSC mixed with hydroxyapatite-tricalcium phosphate granules subcutaneously in 8-week-old female nude mice, and the amount of bone formed was assessed using quantitative histology.
RESULTS
RESULTS
BMS-833923, a SMO antagonist/Hedgehog inhibitor, exhibited significant inhibitory effects on osteoblast differentiation of hMSCs reflected by decreased ALP activity, in vitro mineralization, and downregulation of osteoblast-related gene expression. Similarly, we observed decreased in vivo ectopic bone formation. Global gene expression profiling of BMS-833923-treated compared to vehicle-treated control cells, identified 348 upregulated and 540 downregulated genes with significant effects on multiple signaling pathways, including GPCR, endochondral ossification, RANK-RANKL, insulin, TNF alpha, IL6, and inflammatory response. Further bioinformatic analysis employing Ingenuity Pathway Analysis revealed significant enrichment in BMS-833923-treated cells for a number of functional categories and networks involved in connective and skeletal tissue development and disorders, e.g., NF
CONCLUSIONS
CONCLUSIONS
We identified SMO/Hedgehog antagonist (BMS-833923) as a powerful inhibitor of osteoblastic differentiation of hMSC that may be useful as a therapeutic option for treating conditions associated with high heterotopic bone formation and mineralization.
Identifiants
pubmed: 31871467
doi: 10.1155/2019/3435901
pmc: PMC6907053
doi:
Types de publication
Journal Article
Langues
eng
Pagination
3435901Informations de copyright
Copyright © 2019 Nihal AlMuraikhi et al.
Déclaration de conflit d'intérêts
The authors declare no conflict of interest.
Références
Onco Targets Ther. 2012;5:47-58
pubmed: 22500124
Oncotarget. 2018 Feb 5;9(17):13551-13564
pubmed: 29568377
Bone. 2015 Jan;70:28-36
pubmed: 25138551
Dev Cell. 2008 Apr;14(4):624-32
pubmed: 18410737
Front Oncol. 2019 Sep 18;9:910
pubmed: 31620367
Nature. 2005 Oct 13;437(7061):1032-7
pubmed: 16136080
Wiley Interdiscip Rev Dev Biol. 2014 Nov-Dec;3(6):489-500
pubmed: 25270716
Eur J Clin Nutr. 1994 Feb;48 Suppl 1:S190-7; discussion S198
pubmed: 8005086
Development. 2014 Sep;141(18):3445-57
pubmed: 25183867
Int J Oral Sci. 2015 Jun 26;7(2):73-9
pubmed: 26023726
Int J Immunopathol Pharmacol. 2008 Jul-Sep;21(3):485-91
pubmed: 18831915
J Endocrinol. 2002 Aug;174(2):R1-6
pubmed: 12176676
Cell. 2000 Feb 18;100(4):423-34
pubmed: 10693759
Stem Cells Int. 2018 Feb 15;2018:6913594
pubmed: 29535777
Gene. 2005 Aug 29;357(1):1-8
pubmed: 16125875
Stem Cell Res Ther. 2018 Jan 22;9(1):13
pubmed: 29357923
Nature. 2003 May 15;423(6937):332-6
pubmed: 12748651
Biochem Biophys Res Commun. 2005 Jan 21;326(3):527-38
pubmed: 15596132
Drug Saf. 2019 Feb;42(2):263-279
pubmed: 30649745
Endocr Res. 2014;39(4):144-51
pubmed: 24679227
Nat Biotechnol. 2002 Jun;20(6):592-6
pubmed: 12042863
Science. 2012 Oct 12;338(6104):210-2
pubmed: 23066071
Development. 2007 Sep;134(17):3133-44
pubmed: 17670790
Cell Chem Biol. 2017 Mar 16;24(3):252-280
pubmed: 28286127
Front Biosci. 2007 May 01;12:3068-92
pubmed: 17485283
Genes Dev. 1999 Aug 15;13(16):2072-86
pubmed: 10465785
Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):4048-53
pubmed: 17360475
Methods. 2001 Dec;25(4):402-8
pubmed: 11846609
Regen Ther. 2015 Jan 28;1:57-62
pubmed: 31245441
Curr Biol. 2009 Feb 10;19(3):R125-7
pubmed: 19211052
Arch Biochem Biophys. 2008 May 15;473(2):139-46
pubmed: 18395508
Drug Discov Today. 2014 Jun;19(6):801-8
pubmed: 24252867
Immune Netw. 2018 Feb 07;18(1):e8
pubmed: 29503739
Cell Biol Int. 2012 Apr 1;36(4):349-55
pubmed: 22149964
Stem Cells Int. 2019 Aug 22;2019:3041262
pubmed: 31534459
Ann Saudi Med. 2012 Jan-Feb;32(1):68-77
pubmed: 22156642
PLoS One. 2013 Oct 04;8(10):e76785
pubmed: 24124594
Prog Mol Biol Transl Sci. 2015;133:47-75
pubmed: 26123302
Cancers (Basel). 2016 Feb 15;8(2):null
pubmed: 26891329
Stem Cell Res Ther. 2018 Nov 21;9(1):319
pubmed: 30463599
Curr Osteoporos Rep. 2017 Jun;15(3):126-134
pubmed: 28477234