HIF-1α Stabilization Hampers the Chondrogenic Differentiation of Aged Bone Marrow Mesenchymal Stem Cells by Adenosine A <sub>2A</sub> /A <sub>2B</sub> Receptors Imbalance
Trauma and excessive motion are deleterious to cartilage by triggering HIF-1α stabilization, leading to changes in extracellular adenosine formation via CD73. How these changes affect adenosine A<sub>2A</sub>/A<sub>2B</sub> receptors activation balance in chondrogenesis is unclear. We used bone marrow mesenchymal stem cells (BM-MSCs) from aged women to investigate the impact of HIF-1α overactivation on adenosine formation from ATP breakdown and subsequent A<sub>2A</sub>/A<sub>2B</sub> receptor tone-regulating chondrogenesis. The chondrogenic differentiation of BM-MSCs from 27 postmenopausal (<i>Pm</i>) women was induced for 14 days with or without DMOG, a prolyl-4-hydroxylase (PHD) inhibitor that increases HIF-1α intracellular accumulation. Chondrogenesis was ascertained by the nuclear translocation of SOX9, type II/X collagen, and MMP13 production and by transcriptomic RNAseq analysis. Changes in the density of adenosine receptors, ecto-NTPDases, and CD73 were assessed by immunofluorescence confocal microscopy. The kinetics of ATP hydrolysis and adenosine formation was performed by HPLC. DMOG-induced HIF-1α transcriptional activity increased cartilage damage biomarkers (MMP13 and type X collagen), while decreasing cell viability, SOX9 nuclear translocation, and type II collagen production. DMOG upregulated pro-inflammatory genes and down-regulated chondrogenic gene transcripts determined by RNAseq. HIF-1α stabilization decreased CD39/CD73 amounts and activity, thus reducing adenosine formation. Hence, changes in the A<sub>2A</sub>/A<sub>2B</sub> receptor tone resulted in preferential activation of the SCH 442416-sensitive A<sub>2A</sub> receptor, which is deleterious to the cartilage. In conclusion, HIF-1α overactivation hampers the chondrogenic differentiation of aged BM-MSCs by decreasing adenosine formation from ATP hydrolysis via CD39/CD73, leading to preferential activation of the high-affinity A<sub>2A</sub> versus the chondroprotective A<sub>2B</sub> receptor.
Authors
- Adélio Vilaça (ORCID: https://orcid.org/0000-0001-7470-9168)
- Paulo Correia‐de‐Sá (ORCID: https://orcid.org/0000-0002-6114-9189)
- Jean Sévigny (ORCID: https://orcid.org/0000-0003-2922-1600)
- José Bernardo Noronha‐Matos (ORCID: https://orcid.org/0000-0001-6397-2104)
- Ricardo Martins-Ferreira (ORCID: https://orcid.org/0000-0001-9337-1860)
- Catarina Bessa-Andrês (ORCID: https://orcid.org/0000-0002-5414-2691)
- Rafael Neiva (ORCID: https://orcid.org/0000-0002-2271-7075)
- Rui Pinto-Cardoso (ORCID: https://orcid.org/0000-0001-8196-6162)
- Julie Pelletier (ORCID: https://orcid.org/0000-0003-0461-8126)
- Diogo Nóbrega Catelas (ORCID: https://orcid.org/0000-0002-0885-8947)
- António Oliveira (ORCID: https://orcid.org/0000-0001-6499-9552)
- Flávio Pereira-Costa (ORCID: https://orcid.org/0000-0003-1733-8983)
- Bárbara Guerra-Leal
Institutions
- Universidade do Porto (PT)
- Sociedade Brasileira de Ortopedia e Traumatologia (BR)
- Hospital de Santo António (PT)
- Research Network (United States) (US)
- Université Laval (CA)
Publication Details
- Journal
- ACS Pharmacology & Translational Science
- Published
- 2025-06-24
- DOI
- https://doi.org/10.1021/acsptsci.5c00190
- Citations
- 2
- Primary Topic
- Adenosine and Purinergic Signaling
- Type
- article
- Field-Weighted Citation Impact
- 2.24
Funders
- Universidade do Porto
- Fundação para a Ciência e a Tecnologia
- European Regional Development Fund