Hedgehog Signaling Inhibition by Smoothened Antagonist BMS-833923 Reduces Osteoblast Differentiation and Ectopic Bone Formation of Human Skeletal (Mesenchymal) Stem Cells
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. A Smoothened (SMO) antagonist/Hedgehog inhibitor,...
Saved in:
| Main Authors: | , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2019-01-01
|
| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2019/3435901 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849307642390904832 |
|---|---|
| author | Nihal AlMuraikhi Nuha Almasoud Sarah Binhamdan Ghaydaa Younis Dalia Ali Muthurangan Manikandan Radhakrishnan Vishnubalaji Muhammad Atteya Abdulaziz Siyal Musaad Alfayez Abdullah Aldahmash Moustapha Kassem Nehad M. Alajez |
| author_facet | Nihal AlMuraikhi Nuha Almasoud Sarah Binhamdan Ghaydaa Younis Dalia Ali Muthurangan Manikandan Radhakrishnan Vishnubalaji Muhammad Atteya Abdulaziz Siyal Musaad Alfayez Abdullah Aldahmash Moustapha Kassem Nehad M. Alajez |
| author_sort | Nihal AlMuraikhi |
| collection | DOAJ |
| description | 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. 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. 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., NFκB and STAT signaling. 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. |
| format | Article |
| id | doaj-art-1f652ce2bc424c3288a264907e42aba6 |
| institution | Kabale University |
| issn | 1687-966X 1687-9678 |
| language | English |
| publishDate | 2019-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj-art-1f652ce2bc424c3288a264907e42aba62025-08-20T03:54:42ZengWileyStem Cells International1687-966X1687-96782019-01-01201910.1155/2019/34359013435901Hedgehog Signaling Inhibition by Smoothened Antagonist BMS-833923 Reduces Osteoblast Differentiation and Ectopic Bone Formation of Human Skeletal (Mesenchymal) Stem CellsNihal AlMuraikhi0Nuha Almasoud1Sarah Binhamdan2Ghaydaa Younis3Dalia Ali4Muthurangan Manikandan5Radhakrishnan Vishnubalaji6Muhammad Atteya7Abdulaziz Siyal8Musaad Alfayez9Abdullah Aldahmash10Moustapha Kassem11Nehad M. Alajez12Stem Cell Unit, Department of Anatomy, College of Medicine, King Saud University, Riyadh 11461, Saudi ArabiaStem Cell Unit, Department of Anatomy, College of Medicine, King Saud University, Riyadh 11461, Saudi ArabiaStem Cell Unit, Department of Anatomy, College of Medicine, King Saud University, Riyadh 11461, Saudi ArabiaStem Cell Unit, Department of Anatomy, College of Medicine, King Saud University, Riyadh 11461, Saudi ArabiaMolecular Endocrinology Unit (KMEB), Department of Endocrinology, University Hospital of Odense and University of Southern Denmark, Odense, DenmarkStem Cell Unit, Department of Anatomy, College of Medicine, King Saud University, Riyadh 11461, Saudi ArabiaCancer Research Center, Qatar Biomedical Research Institute (QBRI), Hamad Bin Khalifa University (HBKU), Qatar Foundation (QF), PO Box 34110, Doha, QatarStem Cell Unit, Department of Anatomy, College of Medicine, King Saud University, Riyadh 11461, Saudi ArabiaStem Cell Unit, Department of Anatomy, College of Medicine, King Saud University, Riyadh 11461, Saudi ArabiaStem Cell Unit, Department of Anatomy, College of Medicine, King Saud University, Riyadh 11461, Saudi ArabiaStem Cell Unit, Department of Anatomy, College of Medicine, King Saud University, Riyadh 11461, Saudi ArabiaStem Cell Unit, Department of Anatomy, College of Medicine, King Saud University, Riyadh 11461, Saudi ArabiaCancer Research Center, Qatar Biomedical Research Institute (QBRI), Hamad Bin Khalifa University (HBKU), Qatar Foundation (QF), PO Box 34110, Doha, QatarBackground. 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. 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. 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., NFκB and STAT signaling. 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.http://dx.doi.org/10.1155/2019/3435901 |
| spellingShingle | Nihal AlMuraikhi Nuha Almasoud Sarah Binhamdan Ghaydaa Younis Dalia Ali Muthurangan Manikandan Radhakrishnan Vishnubalaji Muhammad Atteya Abdulaziz Siyal Musaad Alfayez Abdullah Aldahmash Moustapha Kassem Nehad M. Alajez Hedgehog Signaling Inhibition by Smoothened Antagonist BMS-833923 Reduces Osteoblast Differentiation and Ectopic Bone Formation of Human Skeletal (Mesenchymal) Stem Cells Stem Cells International |
| title | Hedgehog Signaling Inhibition by Smoothened Antagonist BMS-833923 Reduces Osteoblast Differentiation and Ectopic Bone Formation of Human Skeletal (Mesenchymal) Stem Cells |
| title_full | Hedgehog Signaling Inhibition by Smoothened Antagonist BMS-833923 Reduces Osteoblast Differentiation and Ectopic Bone Formation of Human Skeletal (Mesenchymal) Stem Cells |
| title_fullStr | Hedgehog Signaling Inhibition by Smoothened Antagonist BMS-833923 Reduces Osteoblast Differentiation and Ectopic Bone Formation of Human Skeletal (Mesenchymal) Stem Cells |
| title_full_unstemmed | Hedgehog Signaling Inhibition by Smoothened Antagonist BMS-833923 Reduces Osteoblast Differentiation and Ectopic Bone Formation of Human Skeletal (Mesenchymal) Stem Cells |
| title_short | Hedgehog Signaling Inhibition by Smoothened Antagonist BMS-833923 Reduces Osteoblast Differentiation and Ectopic Bone Formation of Human Skeletal (Mesenchymal) Stem Cells |
| title_sort | hedgehog signaling inhibition by smoothened antagonist bms 833923 reduces osteoblast differentiation and ectopic bone formation of human skeletal mesenchymal stem cells |
| url | http://dx.doi.org/10.1155/2019/3435901 |
| work_keys_str_mv | AT nihalalmuraikhi hedgehogsignalinginhibitionbysmoothenedantagonistbms833923reducesosteoblastdifferentiationandectopicboneformationofhumanskeletalmesenchymalstemcells AT nuhaalmasoud hedgehogsignalinginhibitionbysmoothenedantagonistbms833923reducesosteoblastdifferentiationandectopicboneformationofhumanskeletalmesenchymalstemcells AT sarahbinhamdan hedgehogsignalinginhibitionbysmoothenedantagonistbms833923reducesosteoblastdifferentiationandectopicboneformationofhumanskeletalmesenchymalstemcells AT ghaydaayounis hedgehogsignalinginhibitionbysmoothenedantagonistbms833923reducesosteoblastdifferentiationandectopicboneformationofhumanskeletalmesenchymalstemcells AT daliaali hedgehogsignalinginhibitionbysmoothenedantagonistbms833923reducesosteoblastdifferentiationandectopicboneformationofhumanskeletalmesenchymalstemcells AT muthuranganmanikandan hedgehogsignalinginhibitionbysmoothenedantagonistbms833923reducesosteoblastdifferentiationandectopicboneformationofhumanskeletalmesenchymalstemcells AT radhakrishnanvishnubalaji hedgehogsignalinginhibitionbysmoothenedantagonistbms833923reducesosteoblastdifferentiationandectopicboneformationofhumanskeletalmesenchymalstemcells AT muhammadatteya hedgehogsignalinginhibitionbysmoothenedantagonistbms833923reducesosteoblastdifferentiationandectopicboneformationofhumanskeletalmesenchymalstemcells AT abdulazizsiyal hedgehogsignalinginhibitionbysmoothenedantagonistbms833923reducesosteoblastdifferentiationandectopicboneformationofhumanskeletalmesenchymalstemcells AT musaadalfayez hedgehogsignalinginhibitionbysmoothenedantagonistbms833923reducesosteoblastdifferentiationandectopicboneformationofhumanskeletalmesenchymalstemcells AT abdullahaldahmash hedgehogsignalinginhibitionbysmoothenedantagonistbms833923reducesosteoblastdifferentiationandectopicboneformationofhumanskeletalmesenchymalstemcells AT moustaphakassem hedgehogsignalinginhibitionbysmoothenedantagonistbms833923reducesosteoblastdifferentiationandectopicboneformationofhumanskeletalmesenchymalstemcells AT nehadmalajez hedgehogsignalinginhibitionbysmoothenedantagonistbms833923reducesosteoblastdifferentiationandectopicboneformationofhumanskeletalmesenchymalstemcells |