DLK1 and DLK2, two non-canonical ligands of NOTCH receptors, differentially modulate the osteogenic differentiation of mesenchymal C3H10T1/2 cells.


Journal

Biological research
ISSN: 0717-6287
Titre abrégé: Biol Res
Pays: England
ID NLM: 9308271

Informations de publication

Date de publication:
30 Oct 2024
Historique:
received: 27 08 2024
accepted: 25 10 2024
medline: 30 10 2024
pubmed: 30 10 2024
entrez: 30 10 2024
Statut: epublish

Résumé

C3H10T1/2 is a mesenchymal cell line capable of differentiating into osteoblasts, adipocytes and chondrocytes. The differentiation of these cells into osteoblasts is modulated by various transcription factors, such as RUNX2. Additionally, several interconnected signaling pathways, including the NOTCH pathway, play a crucial role in modulating their differentiation into mature bone cells. We have investigated the roles of DLK1 and DLK2, two non-canonical inhibitory ligands of NOTCH receptors, in the osteogenic differentiation of C3H10T1/2 cells. Our results corroborate existing evidence that DLK1 acts as an inhibitor of osteogenesis. In contrast, we demonstrate for the first time that DLK2 enhances this differentiation process. Additionally, our data suggest that NOTCH2, 3 and 4 receptors may promote osteogenesis, as indicated by their increased expression during this process, whereas NOTCH1 expression, which decreases during cell differentiation, might inhibit osteogenesis. Moreover, treatment with DAPT, a NOTCH signaling inhibitor, impeded osteogenic differentiation. We have confirmed the increase in ERK1/2 MAPK and p38 MAPK phosphorylation in C3H10T1/2 cells induced to differentiate to osteoblasts. Our new findings reveal increased ERK1/2 MAPK phosphorylation in differentiated C3H10T1/2 cells with a decrease in DLK1 expression or an overexpression of DLK2, which is coincident with the behavior of those transfectants where we have detected an increase in osteogenic differentiation. Additionally, p38 MAPK phosphorylation increases in differentiated C3H10T1/2 cells with reduced DLK1 levels. Our results suggest that DLK1 may inhibit osteogenesis, while DLK2 may promote it, by modulating NOTCH signaling and the phosphorylation of ERK1/2 and p38 MAPK pathways. Given the established inhibitory effect of DLK proteins on NOTCH signaling, these new insights could pave the way for developing future therapeutic strategies aimed at treating bone diseases.

Sections du résumé

BACKGROUND BACKGROUND
C3H10T1/2 is a mesenchymal cell line capable of differentiating into osteoblasts, adipocytes and chondrocytes. The differentiation of these cells into osteoblasts is modulated by various transcription factors, such as RUNX2. Additionally, several interconnected signaling pathways, including the NOTCH pathway, play a crucial role in modulating their differentiation into mature bone cells. We have investigated the roles of DLK1 and DLK2, two non-canonical inhibitory ligands of NOTCH receptors, in the osteogenic differentiation of C3H10T1/2 cells.
RESULTS RESULTS
Our results corroborate existing evidence that DLK1 acts as an inhibitor of osteogenesis. In contrast, we demonstrate for the first time that DLK2 enhances this differentiation process. Additionally, our data suggest that NOTCH2, 3 and 4 receptors may promote osteogenesis, as indicated by their increased expression during this process, whereas NOTCH1 expression, which decreases during cell differentiation, might inhibit osteogenesis. Moreover, treatment with DAPT, a NOTCH signaling inhibitor, impeded osteogenic differentiation. We have confirmed the increase in ERK1/2 MAPK and p38 MAPK phosphorylation in C3H10T1/2 cells induced to differentiate to osteoblasts. Our new findings reveal increased ERK1/2 MAPK phosphorylation in differentiated C3H10T1/2 cells with a decrease in DLK1 expression or an overexpression of DLK2, which is coincident with the behavior of those transfectants where we have detected an increase in osteogenic differentiation. Additionally, p38 MAPK phosphorylation increases in differentiated C3H10T1/2 cells with reduced DLK1 levels.
CONCLUSIONS CONCLUSIONS
Our results suggest that DLK1 may inhibit osteogenesis, while DLK2 may promote it, by modulating NOTCH signaling and the phosphorylation of ERK1/2 and p38 MAPK pathways. Given the established inhibitory effect of DLK proteins on NOTCH signaling, these new insights could pave the way for developing future therapeutic strategies aimed at treating bone diseases.

Identifiants

pubmed: 39473022
doi: 10.1186/s40659-024-00561-7
pii: 10.1186/s40659-024-00561-7
doi:

Substances chimiques

Intercellular Signaling Peptides and Proteins 0
Calcium-Binding Proteins 0
Receptors, Notch 0
Dlk1 protein, mouse 0
Dlk2 protein, mouse 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

77

Subventions

Organisme : The Health Council of the Regional Government of Castilla-La Mancha (Spain), supported by the Fondo Europeo de Desarrollo Regional (FEDER).
ID : ISBPLY/17/180501/000316

Informations de copyright

© 2024. The Author(s).

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Auteurs

María-Milagros Rodríguez-Cano (MM)

Biochemistry and Molecular Biology Branch, Department of Inorganic, Organic Chemistry and Biochemistry, Pharmacy School/IB-UCLM/Biomedicine Unit, University of Castilla-La Mancha/CSIC, Albacete, Spain.

María-Julia González-Gómez (MJ)

Biochemistry and Molecular Biology Branch, Department of Inorganic, Organic Chemistry and Biochemistry, ETSIAMB/IB-UCLM/Biomedicine Unit, University of Castilla-La Mancha/CSIC, Albacete, Spain.

Eva-María Monsalve (EM)

Biochemistry and Molecular Biology Branch, Department of Inorganic, Organic Chemistry and Biochemistry, Medical School/IB-UCLM/Biomedicine Unit, University of Castilla-La Mancha/CSIC, Albacete, Spain.

María-José M Díaz-Guerra (MM)

Biochemistry and Molecular Biology Branch, Department of Inorganic, Organic Chemistry and Biochemistry, Medical School/IB-UCLM/Biomedicine Unit, University of Castilla-La Mancha/CSIC, Albacete, Spain.

Moustapha Kassem (M)

Molecular Endocrinology Unit (KMEB), Department of Endocrinology and Metabolism, University Hospital of Odense and Danish Institute for Advanced Study, University of Southern Denmark, Odense, Denmark.

Jorge Laborda (J)

Biochemistry and Molecular Biology Branch, Department of Inorganic, Organic Chemistry and Biochemistry, Pharmacy School/IB-UCLM/Biomedicine Unit, University of Castilla-La Mancha/CSIC, Albacete, Spain.

María-Luisa Nueda (ML)

Biochemistry and Molecular Biology Branch, Department of Inorganic, Organic Chemistry and Biochemistry, Pharmacy School/IB-UCLM/Biomedicine Unit, University of Castilla-La Mancha/CSIC, Albacete, Spain. MariaLuisa.Nueda@uclm.es.

Victoriano Baladrón (V)

Biochemistry and Molecular Biology Branch, Department of Inorganic, Organic Chemistry and Biochemistry, Medical School/IB-UCLM/Biomedicine Unit, University of Castilla-La Mancha/CSIC, Albacete, Spain. Victoriano.Baladron@uclm.es.

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