Synergistic effects of CoCl2 and ROCK inhibition on mesenchymal stem cell differentiation into neuron-like cells

• DOI: 10.1242/jcs.03004
• Publication Date: 2006-06-14T03:44:09Z
• Authors: Emilie Pacary; Hélène Legros; Samuel Valable; Pascal Duchatelle; Myriam Lecocq; Edwige Petit; Olivier Nicole; Myriam Bernaudin
• Keywords: Male; 0301 basic medicine; Pyridines; Down-Regulation; Bone Marrow Cells; Echinomycin; Protein Serine-Threonine Kinases; Mice; 03 medical and health sciences; Animals; [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]; Cell Proliferation; Neurons; rho-Associated Kinases; 0303 health sciences; Cell Cycle; Intracellular Signaling Peptides and Proteins; Cell Differentiation; Drug Synergism; Mesenchymal Stem Cells; Cobalt; Hypoxia-Inducible Factor 1, alpha Subunit; Amides; 3. Good health; Gene Expression Regulation; [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]

Bone-marrow-derived mesenchymal stem cells (MSCs) constitute an interesting cellular source to promote brain regeneration after neurodegenerative diseases. Recently, several studies suggested that oxygen-dependent gene expression is of crucial importance in governing the essential steps of neurogenesis such as cell proliferation, survival and differentiation. In this context, we analysed the effect of the HIF-1 (hypoxia inducible factor-1) activation-mimicking agent CoCl2 on MSCs. CoCl2 treatment increased the expression of the anti-proliferative gene BTG2/PC3 and decreased cyclin D1 expression. Expression of HIF-1α and its target genes EPO, VEGF and p21 was also upregulated. These changes were followed by inhibition of cell proliferation and morphological changes resulting in neuron-like cells, which had increased neuronal marker expression and responded to neurotransmitters. Echinomycin, a molecule inhibiting HIF-1 DNA-binding activity, blocked the CoCl2 effect on MSCs. Additionally, by using Y-27632, we demonstrated that Rho kinase (ROCK) inhibition potentiated CoCl2-induced MSC differentiation in particular into dopaminergic neuron-like cells as attested by its effect on tyrosine hydroxylase expression. Altogether, these results support the ability of MSCs to differentiate into neuron-like cells in response to CoCl2, an effect that might act, in part, through HIF-1 activation and cell-cycle arrest, and which is potentiated by inhibition of ROCK.