Abstract
We sought to address the fundamental question of how stem cell microenvironments can regulate self-renewal. We found that Notch was active in astroglia-like neural stem cells (NSCs), but not in transit-amplifying progenitors of the murine subependymal zone, and that the level of Notch transcriptional activity correlated with self-renewal and multipotency. Moreover, dividing NSCs appeared to balance renewal with commitment via controlled segregation of Notch activity, leading to biased expression of known (Hes1) and previously unknown (Egfr) Notch target genes in daughter cells. Pigment epithelium–derived factor (PEDF) enhanced Notch-dependent transcription in cells with low Notch signaling, thereby subverting the output of an asymmetrical division to the production of two highly self-renewing cells. Mechanistically, PEDF induced a non-canonical activation of the NF-κB pathway, leading to the dismissal of the transcriptional co-repressor N-CoR from specific Notch-responsive promoters. Our data provide a basis for stemness regulation in vascular niches and indicate that Notch and PEDF cooperate to regulate self-renewal.
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Acknowledgements
We are grateful to A. Bigas, J.L. de la Pompa, N. Gaiano, W.C. Greene, O. Hermanson, S. Hitoshi, A. Israel, K. Jepsen, M. Karin, R. Kopan, G. Leclercq, M.G. Rosenfeld, S. Sun, D. van der Kooy, Y. Zhan and V. Meni for kindly providing constructs and reagents. We also thank E. Porlan and H. Mira for technical advice, M.P. Rubio for excellent technical assistance, and E. Porlan and S.R. Ferrón for critical reading of the manuscript and valuable discussions. We gratefully acknowledge the help of M.A. Marqués-Torrejón with the infusion experiments and of A. Martínez and D. Gil with cytometry. We are also grateful to E. Lai for comments on the manuscript. This work was supported by grants from Ministerio de Ciencia e Innovación (SAF Program), Ministerio de Sanidad (Red Tercel, Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas, CIBERNED), Generalitat Valenciana (Programa Prometeo) and Fundación 'la Caixa'. C.A.-A. was a recipient of a Formación del Profesorado Universitario predoctoral fellowship. J.M.M.-R. and A.C.D. were funded by CIBERNED.
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All of the authors designed and discussed the experiments. C.A.-A. conducted most of the cell culture and biochemistry experiments on the relationship between PEDF and Notch, Notch/EGFR asymmetry and the role of N-CoR and p65, as well as the infusion experiments and in vivo analyses. C.A.-A. and J.M.M.-R. performed the FACS experiments and analyses with TNR cells, ChIP studies and luciferase assays. A.C.D. contributed to immunohistochemistry analyses and multipotency assays. I.F. supervised the project and wrote the manuscript.
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The authors have an active patent on the use of PEDF for stem-cell renewal.
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Andreu-Agulló, C., Morante-Redolat, J., Delgado, A. et al. Vascular niche factor PEDF modulates Notch-dependent stemness in the adult subependymal zone. Nat Neurosci 12, 1514–1523 (2009). https://doi.org/10.1038/nn.2437
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DOI: https://doi.org/10.1038/nn.2437
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