1. Université Côte d'Azur, INSERM, CNRS, IPMC, team labeled “Laboratory of Excellence (LABEX) Distalz”, 660 route des Lucioles, 06560, Sophia-Antipolis, Valbonne, France.
2. Department of Biology, The University of North Carolina Greensboro, Greensboro, North Carolina, United States of America.
3. Department of Life Sciences, University of Trieste, 34127 Trieste, Italy.
4. AP-HP, Service de Neurologie 2 - Mazarin, Groupe Hospitalier Pitié-Salpêtrière, Université Pierre & Marie Curie Paris VI, Centre de Recherche de l'Institut du Cerveau et de la Moelle Epinière, UMRS 975, Paris, France.
*Co-first authors with equal contributions to this work.
#Co-second authors with equal contributions to this work.
Parkin (PK) is an E3-ligase harboring tumor suppressor properties that has been associated to various cancer types including glioblastoma (GBM). However, PK is also a transcription factor (TF), the contribution of which to GBM etiology remains to be established.
Methods: The impact of PK on GBM cells proliferation was analyzed by real-time impedance measurement and flow cytometry. Cyclins A and B proteins, promoter activities and mRNA levels were measured by western blot, luciferase assay and quantitative real-time PCR. Protein-protein and protein-promoter interactions were performed by co-immunoprecipitation and by ChIP approaches. The contribution of endogenous PK to tumor progression in vivo was performed by allografts of GL261 GBM cells in wild-type and PK knockout mice.
Results: We show that overexpressed and endogenous PK control GBM cells proliferation by modulating the S and G2/M phases of the cell cycle via the trans-repression of cyclin A and cyclin B genes. We establish that cyclin B is regulated by both E3-ligase and TF PK functions while cyclin A is exclusively regulated by PK TF function. PK invalidation leads to enhanced tumor progression in immunocompetent mice suggesting an impact of PK-dependent tumor environment to tumor development. We show that PK is secreted by neuronal cells and recaptured by tumor cells. Recaptured PK lowered cyclins levels and decreased GBM cells proliferation. Further, PK expression is decreased in human GBM biopsies and its expression is inversely correlated to both cyclins A and B expressions.
Conclusion: Our work demonstrates that PK tumor suppressor function contributes to the control of tumor by its cellular environment. It also shows a key role of PK TF function in GBM development via the control of cyclins in vitro and in vivo. It suggests that therapeutic strategies aimed at controlling PK shuttling to the nucleus may prove useful to treat GBM.
Keywords: glioblastoma, parkin, cyclins, proliferation, transcription factor