Synergistic effects of CoCl(2) and ROCK inhibition on mesenchymal stem cell differentiation into neuron-like cells.
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 CoCl(2) on MSCs. CoCl(2) treatment increased the expression of the anti-proliferative gene BTG2/PC3 and decreased cyclin D1 expression. Expression of HIF-1alpha 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 CoCl(2) effect on MSCs. Additionally, by using Y-27632, we demonstrated that Rho kinase (ROCK) inhibition potentiated CoCl(2)-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 CoCl(2), an effect that might act, in part, through HIF-1 activation and cell-cycle arrest, and which is potentiated by inhibition of ROCK.
Pacary
Emilie
E
UMR-CNRS 6185, Neurodegenerescence: models and therapeutic strategies, University of Caen, CYCERON, Bd Henri Becquerel, BP 5229, 14074 Caen CEDEX, France.
Legros
Hélène
H
Valable
Samuel
S
Duchatelle
Pascal
P
Lecocq
Myriam
M
Petit
Edwige
E
Nicole
Olivier
O
Bernaudin
Myriam
M
D000577:Amides / Q000494:pharmacology; D000818:Animals; D001854:Bone Marrow Cells / Q000187:drug effects; D002453:Cell Cycle / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D003035:Cobalt / Q000494:pharmacology*; D015536:Down-Regulation; D004357:Drug Synergism; D004448:Echinomycin / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D008297:Male; D059630:Mesenchymal Stem Cells / Q000187:drug effects*; D051379:Mice; D009474:Neurons / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D011725:Pyridines / Q000494:pharmacology; D054460:rho-Associated Kinases
D000577:Amides; C497443:Hif1a protein, mouse; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D047908:Intracellular Signaling Peptides and Proteins; D011725:Pyridines; C108830:Y 27632; D003035:Cobalt; D004448:Echinomycin; D017346:Protein Serine-Threonine Kinases; D054460:rho-Associated Kinases; C018021:cobaltous chloride