G-quadruplex ligands exert their antiproliferative effects through telomere-dependent and telomere-independent mechanisms, but the inter-relationships among autophagy, cell growth arrest and cell death induced by these ligands remain largely unexplored. Here, we demonstrate that the G-quadruplex ligand 20A causes growth arrest of cancer cells in culture and in a HeLa cell xenografted mouse model. This response is associated with the induction of senescence and apoptosis. Transcriptomic analysis of 20A treated cells reveals a significant functional enrichment of biological pathways related to growth arrest, DNA damage response and the lysosomal pathway. 20A elicits global DNA damage but not telomeric damage and activates the ATM and autophagy pathways. Loss of ATM following 20A treatment inhibits both autophagy and senescence and sensitizes cells to death. Moreover, disruption of autophagy by deletion of two essential autophagy genes ATG5 and ATG7 leads to failure of CHK1 activation by 20A and subsequently increased cell death. Our results, therefore, identify the activation of ATM by 20A as a critical player in the balance between senescence and apoptosis and autophagy as one of the key mediators of such regulation. Thus, targeting the ATM/autophagy pathway might be a promising strategy to achieve the maximal anticancer effect of this compound.

ARNA Laboratory Université de Bordeaux INSERM U1212 CNRS UMR 5320 IECB F 33600 Pessac France

Cancer Research UK Beatson Institute Glasgow G611BD UK and Institute of Cancer Sciences University of Glasgow Glasgow G61 1QH UK

Centre de Bioinformatique de Bordeaux université de Bordeaux F 33000 Bordeaux France

Department of Biopathology Institut Bergonié F 33076 Bordeaux France

Inserm U1065 C3M Team Myeloid Malignancies and Multiple Myeloma Université Côte d'Azur F 06204 Nice France

Institut Bergonié Université de Bordeaux INSERM U1218 F 33076 Bordeaux France

Institut Curie PSL Research University CNRS UMR3244 F 75005 Paris France

Institute of Biophysics of the Czech Academy of Sciences Královopolská 135 612 65 Brno Czech Republic

Service commun des animaleries Université de Bordeaux F 33000 Bordeaux France

Sorbonne Universités UPMC Univ Paris 06 CNRS UMR3244 F 75005 Paris France

Université Côte d'Azur Centre Commun de Microscopie Appliquée 06108 Nice France

Université de Bordeaux Centre de Génomique Fonctionnelle Plateforme Protéome F 33000 Bordeaux France

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G-quadruplex ligands as potent regulators of lysosomes

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1

Activation of the Ataxia Telangiectasia Mutated/Autophagy pathway by a G-quadruplex ligand links senescence with apoptosis