The CREB1 inhibitor 666-15 maintains cartilage homeostasis and mitigates osteoarthritis progression
Aims: cAMP response element binding protein (CREB1) is involved in the progression of osteoarthritis (OA). However, available findings about the role of CREB1 in OA are inconsistent. 666-15 is a potent and selective CREB1 inhibitor, but its role in OA is unclear. This study aimed to investigate the...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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The British Editorial Society of Bone & Joint Surgery
2024-01-01
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| Series: | Bone & Joint Research |
| Subjects: | |
| Online Access: | https://online.boneandjoint.org.uk/doi/epdf/10.1302/2046-3758.131.BJR-2023-0016.R2 |
| _version_ | 1797354194727862272 |
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| author | Ying Wang Zhimin Wu Guoqiang Yan Shan Li Yanzhuo Zhang Guangping Li Chengai Wu |
| author_facet | Ying Wang Zhimin Wu Guoqiang Yan Shan Li Yanzhuo Zhang Guangping Li Chengai Wu |
| author_sort | Ying Wang |
| collection | DOAJ |
| description | Aims: cAMP response element binding protein (CREB1) is involved in the progression of osteoarthritis (OA). However, available findings about the role of CREB1 in OA are inconsistent. 666-15 is a potent and selective CREB1 inhibitor, but its role in OA is unclear. This study aimed to investigate the precise role of CREB1 in OA, and whether 666-15 exerts an anti-OA effect. Methods: CREB1 activity and expression of a disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS4) in cells and tissues were measured by immunoblotting and immunohistochemical (IHC) staining. The effect of 666-15 on chondrocyte viability and apoptosis was examined by cell counting kit-8 (CCK-8) assay, JC-10, and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling (TUNEL) staining. The effect of 666-15 on the microstructure of subchondral bone, and the synthesis and catabolism of cartilage, in anterior cruciate ligament transection mice were detected by micro-CT, safranin O and fast green (S/F), immunohistochemical staining, and enzyme-linked immunosorbent assay (ELISA). Results: CREB1 was hyperactive in osteoarthritic articular cartilage, interleukin (IL)-1β-treated cartilage explants, and IL-1β- or carbonyl cyanide 3-chlorophenylhydrazone (CCCP)-treated chondrocytes. 666-15 enhanced cell viability of OA-like chondrocytes and alleviated IL-1β- or CCCP-induced chondrocyte injury through inhibition of mitochondrial dysfunction-associated apoptosis. Moreover, inhibition of CREB1 by 666-15 suppressed expression of ADAMTS4. Additionally, 666-15 alleviated joint degeneration in an ACLT mouse model. Conclusion: Hyperactive CREB1 played a critical role in OA development, and 666-15 exerted anti-IL-1β or anti-CCCP effects in vitro as well as joint-protective effects in vivo. 666-15 may therefore be used as a promising anti-OA drug. Cite this article: Bone Joint Res 2024;13(1):4–18. |
| first_indexed | 2024-03-08T13:45:58Z |
| format | Article |
| id | doaj.art-028c3b560d3140efbab9e8b1725b543c |
| institution | Directory Open Access Journal |
| issn | 2046-3758 |
| language | English |
| last_indexed | 2024-03-08T13:45:58Z |
| publishDate | 2024-01-01 |
| publisher | The British Editorial Society of Bone & Joint Surgery |
| record_format | Article |
| series | Bone & Joint Research |
| spelling | doaj.art-028c3b560d3140efbab9e8b1725b543c2024-01-16T08:40:31ZengThe British Editorial Society of Bone & Joint SurgeryBone & Joint Research2046-37582024-01-0113141810.1302/2046-3758.131.BJR-2023-0016.R2The CREB1 inhibitor 666-15 maintains cartilage homeostasis and mitigates osteoarthritis progressionYing Wang0https://orcid.org/0000-0003-2873-7197Zhimin Wu1https://orcid.org/0000-0003-2555-6403Guoqiang Yan2Shan Li3https://orcid.org/0000-0002-0272-6193Yanzhuo Zhang4Guangping Li5Chengai Wu6https://orcid.org/0000-0002-6366-4758Department of Molecular Orthopedics, National Center for Orthopaedics, Beijing Research Institute of Traumatology and Orthopaedics, Beijing Jishuitan Hospital, Capital Medical University, Beijing, ChinaDepartment of Molecular Orthopedics, National Center for Orthopaedics, Beijing Research Institute of Traumatology and Orthopaedics, Beijing Jishuitan Hospital, Capital Medical University, Beijing, ChinaNational Center for Orthopaedics, Animal Laboratory, Beijing Research Institute of Traumatology and Orthopaedics, Beijing Jishuitan Hospital, Capital Medical University, Beijing, ChinaDepartment of Molecular Orthopedics, National Center for Orthopaedics, Beijing Research Institute of Traumatology and Orthopaedics, Beijing Jishuitan Hospital, Capital Medical University, Beijing, ChinaDepartment of Molecular Orthopedics, National Center for Orthopaedics, Beijing Research Institute of Traumatology and Orthopaedics, Beijing Jishuitan Hospital, Capital Medical University, Beijing, ChinaNational Center for Orthopaedics, Laboratory of Bone Tissue Engineering, Beijing Research Institute of Traumatology and Orthopaedics, Beijing Jishuitan Hospital, Capital Medical University, Beijing, ChinaDepartment of Molecular Orthopedics, National Center for Orthopaedics, Beijing Research Institute of Traumatology and Orthopaedics, Beijing Jishuitan Hospital, Capital Medical University, Beijing, ChinaAims: cAMP response element binding protein (CREB1) is involved in the progression of osteoarthritis (OA). However, available findings about the role of CREB1 in OA are inconsistent. 666-15 is a potent and selective CREB1 inhibitor, but its role in OA is unclear. This study aimed to investigate the precise role of CREB1 in OA, and whether 666-15 exerts an anti-OA effect. Methods: CREB1 activity and expression of a disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS4) in cells and tissues were measured by immunoblotting and immunohistochemical (IHC) staining. The effect of 666-15 on chondrocyte viability and apoptosis was examined by cell counting kit-8 (CCK-8) assay, JC-10, and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling (TUNEL) staining. The effect of 666-15 on the microstructure of subchondral bone, and the synthesis and catabolism of cartilage, in anterior cruciate ligament transection mice were detected by micro-CT, safranin O and fast green (S/F), immunohistochemical staining, and enzyme-linked immunosorbent assay (ELISA). Results: CREB1 was hyperactive in osteoarthritic articular cartilage, interleukin (IL)-1β-treated cartilage explants, and IL-1β- or carbonyl cyanide 3-chlorophenylhydrazone (CCCP)-treated chondrocytes. 666-15 enhanced cell viability of OA-like chondrocytes and alleviated IL-1β- or CCCP-induced chondrocyte injury through inhibition of mitochondrial dysfunction-associated apoptosis. Moreover, inhibition of CREB1 by 666-15 suppressed expression of ADAMTS4. Additionally, 666-15 alleviated joint degeneration in an ACLT mouse model. Conclusion: Hyperactive CREB1 played a critical role in OA development, and 666-15 exerted anti-IL-1β or anti-CCCP effects in vitro as well as joint-protective effects in vivo. 666-15 may therefore be used as a promising anti-OA drug. Cite this article: Bone Joint Res 2024;13(1):4–18.https://online.boneandjoint.org.uk/doi/epdf/10.1302/2046-3758.131.BJR-2023-0016.R2creb1666-15cartilage homeostasisosteoarthritisosteoarthritis (oa)cartilage homeostasischondrocytescartilage tissuesanterior cruciate ligament transection (aclt)stainingarticular cartilage tissueapoptosissubchondral bonemouse model |
| spellingShingle | Ying Wang Zhimin Wu Guoqiang Yan Shan Li Yanzhuo Zhang Guangping Li Chengai Wu The CREB1 inhibitor 666-15 maintains cartilage homeostasis and mitigates osteoarthritis progression Bone & Joint Research creb1 666-15 cartilage homeostasis osteoarthritis osteoarthritis (oa) cartilage homeostasis chondrocytes cartilage tissues anterior cruciate ligament transection (aclt) staining articular cartilage tissue apoptosis subchondral bone mouse model |
| title | The CREB1 inhibitor 666-15 maintains cartilage homeostasis and mitigates osteoarthritis progression |
| title_full | The CREB1 inhibitor 666-15 maintains cartilage homeostasis and mitigates osteoarthritis progression |
| title_fullStr | The CREB1 inhibitor 666-15 maintains cartilage homeostasis and mitigates osteoarthritis progression |
| title_full_unstemmed | The CREB1 inhibitor 666-15 maintains cartilage homeostasis and mitigates osteoarthritis progression |
| title_short | The CREB1 inhibitor 666-15 maintains cartilage homeostasis and mitigates osteoarthritis progression |
| title_sort | creb1 inhibitor 666 15 maintains cartilage homeostasis and mitigates osteoarthritis progression |
| topic | creb1 666-15 cartilage homeostasis osteoarthritis osteoarthritis (oa) cartilage homeostasis chondrocytes cartilage tissues anterior cruciate ligament transection (aclt) staining articular cartilage tissue apoptosis subchondral bone mouse model |
| url | https://online.boneandjoint.org.uk/doi/epdf/10.1302/2046-3758.131.BJR-2023-0016.R2 |
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