, The number of Galectin3 puncta within each cell was scored using the "Find Maxima" tool

, To assess lysosomal membrane permeabilization (LMP), co-localization study of Galectin3 puncta and LAMP1 was carried out by immunofluorescence analysis. Cells were grown on glass coverslips, fixed in ice-cold methanol at ?20 ? C for 15 min and then blocked/permeabilized with 3% BSA containing

, After washing, cells were incubated with the secondary antibody (anti-rabbit Alexa Fluor 488 antibody) for 1 h at room temperature in a dark chamber and stained with 2 µg/mL of Hoechst 33258 (H3569, Triton X-100 for 1 h. Cells were then incubated with the primary antibody overnight at 4 ? C

, For some experiments, cells were stained with 150 nM tetramethylrhodamine methyl ester (TMRM, from Molecular Probes-Life Technologies) and 5 µg/mL 4 ,6-diaminidino-2-phenylindole (DAPI, from Molecular Probes-Life Technologies) for 30 min at 37 ? C. Cytofluorometric determinations of early and late apoptotis were carried out on a MACSQuant cytometer, Cell Death Cell death was determined by measuring the plasma membrane permeability using propidium iodide dye (P4864, Sigma). Briefly, supernatant and attached cells were collected, vol.93482, 1862495.

, Cell viability Assay MTT assay was used to monitor cell viability in cells subjected to different treatments, Briefly, cells

M. Reader, To determine the IC50 of 20A derivative compounds, a non-linear regression curve was fit using GraphPad Prism software, Molecular Devices

J. L. and M. D. , A-V is supported by the Mexican National Council of Science and Technology (CONACYT, funding #757821), Ligue Régionale d'Aquitaine, (to M.D-M). K

, Agence National de la Recherche (ANR) -Projets blancs

, Flash-covid-19" LYSOMOTROP; the ERA-Net for Research on Rare Diseases

U. Ammica and . Cnrs-ums3655, Association pour la recherche sur le cancer (ARC)

, Association, Le Cancer du Sein, Parlons-en!

C. Ile-de-france,

, Fondation pour la Recherche Médicale (FRM); a donation by Elior

, European Research Area Network on Cardiovascular Diseases (ERA-CVD

, Gustave Roussy Odyssea, the European Union Horizon 2020 Project Oncobiome; Fondation Carrefour; High-end Foreign Expert Program in China (GDW20171100085)

, Institut National du Cancer

. Inserm, HTE

, LeDucq Foundation; the LabEx Immuno-Oncology (ANR-18-IDEX-0001); the RHU Torino Lumière; the Seerave Foundation; the SIRIC Stratified Oncology Cell DNA Repair and Tumor Immune Elimination (SOCRATE); and the SIRIC Cancer Research and Personalized Medicine (CARPEM)

H. Xu and D. Ren, Lysosomal physiology, Annu. Rev. Physiol, vol.77, pp.57-80, 2015.

H. Appelqvist, P. Wäster, K. Kågedal, and K. Öllinger, The lysosome: From waste bag to potential therapeutic target, J. Mol. Cell Biol, vol.5, pp.214-226, 2013.

G. Kroemer and M. Jäättelä, Lysosomes and autophagy in cell death control, Nat. Rev. Cancer, vol.5, pp.886-897, 2005.

N. Mizushima, Autophagy: Process and function, Genes Dev, vol.21, pp.2861-2873, 2007.

P. Boya, Lysosomal function and dysfunction: Mechanism and disease, Antioxid. Redox Signal, vol.17, pp.766-774, 2012.

T. Kallunki, O. D. Olsen, and M. Jäättelä, Cancer-associated lysosomal changes: Friends or foes? Oncogene, vol.32, 1995.

Y. Gong, M. Duvvuri, M. B. Duncan, J. Liu, and J. P. Krise, Niemann-Pick C1 protein facilitates the efflux of the anticancer drug daunorubicin from cells according to a novel vesicle-mediated pathway, J. Pharmacol. Exp. Ther, vol.316, pp.242-247, 2006.

M. Herlevsen, G. Oxford, C. R. Owens, M. Conaway, and D. Theodorescu, Depletion of major vault protein increases doxorubicin sensitivity and nuclear accumulation and disrupts its sequestration in lysosomes, Mol. Cancer Ther, vol.6, pp.1804-1813, 2007.

P. J. Smith, H. R. Sykes, M. E. Fox, and I. J. Furlong, Subcellular distribution of the anticancer drug mitoxantrone in human and drug-resistant murine cells analyzed by flow cytometry and confocal microscopy and its relationship to the induction of DNA damage, Cancer Res, vol.52, pp.4000-4008, 1992.

B. Zhitomirsky and Y. G. Assaraf, Lysosomal sequestration of hydrophobic weak base chemotherapeutics triggers lysosomal biogenesis and lysosome-dependent cancer multidrug resistance, Oncotarget, vol.6, pp.1143-1156, 2015.

S. D. Goldman, R. S. Funk, R. A. Rajewski, and J. P. Krise, Mechanisms of amine accumulation in, and egress from, lysosomes, Bioanalysis, vol.1, pp.1445-1459, 2009.

P. Ferrao, P. Sincock, S. Cole, and L. Ashman, Intracellular P-gp contributes to functional drug efflux and resistance in acute myeloid leukaemia, Leuk. Res, vol.25, pp.395-405, 2001.

U. Repnik, M. Hafner?esen, and B. Turk, Lysosomal membrane permeabilization in cell death: Concepts and challenges. Mitochondrion, vol.19, pp.49-57, 2014.

N. Fehrenbacher, M. Gyrd-hansen, B. Poulsen, U. Felbor, T. Kallunki et al., Sensitization to the lysosomal cell death pathway upon immortalization and transformation, Cancer Res, vol.64, pp.5301-5310, 2004.

N. H. Petersen, O. D. Olsen, L. Groth-pedersen, A. Ellegaard, M. Bilgin et al., Transformation-associated changes in sphingolipid metabolism sensitize cells to lysosomal cell death induced by inhibitors of acid sphingomyelinase, Cancer Cell, vol.24, pp.379-393, 2013.

P. Boya, K. Andreau, D. Poncet, N. Zamzami, J. Perfettini et al., Lysosomal membrane permeabilization induces cell death in a mitochondrion-dependent fashion, J. Exp. Med, vol.197, pp.1323-1334, 2003.

S. Aits and M. Jäättelä, Lysosomal cell death at a glance, J. Cell Sci, vol.126, 1905.

S. Piao and R. K. Amaravadi, Targeting the lysosome in cancer, Ann. N. Y. Acad. Sci, p.45, 1371.

C. K. Kwok and C. J. Merrick, Quadruplexes: Prediction, characterization, and biological application, Trends Biotechnol, vol.35, pp.997-1013, 2017.

I. M. Dixon, F. Lopez, A. M. Tejera, J. Estève, M. A. Balasco et al., A G-Quadruplex ligand with 10000-fold selectivity over duplex DNA, J. Am. Chem. Soc, vol.129, pp.1502-1503, 2007.

J. M. Nicoludis, S. P. Barrett, J. L. Mergny, and L. A. Yatsunyk, Interaction of human telomeric DNA with N-methyl mesoporphyrin IX, Nucleic Acids Res, vol.40, pp.5432-5447, 2012.

A. D'urso, R. Randazzo, V. Rizzo, C. Gangemi, V. Romanucci et al., Stabilization vs. destabilization of G-quadruplex superstructures: The role of the porphyrin derivative having spermine arms, Phys. Chem. Chem. Phys, vol.19, pp.17404-17410, 2017.

G. Cimino-reale, N. Zaffaroni, and M. Folini, Emerging role of G-quadruplex DNA as target in anticancer therapy, Curr. Pharm. Des, vol.22, pp.6612-6624, 2016.

J. Beauvarlet, P. Bensadoun, E. Darbo, G. Labrunie, B. Rousseau et al., Modulation of the ATM/autophagy pathway by a G-quadruplex ligand tips the balance between senescence and apoptosis in cancer cells, Nucleic Acids Res, vol.47, pp.2739-2756, 2019.
URL : https://hal.archives-ouvertes.fr/hal-02038474

, Cancers, vol.12, 2020.

N. M. Smith, G. Labrunie, B. Corry, P. L. Tran, M. Norret et al., Unraveling the relationship between structure and stabilization of triarylpyridines as G-quadruplex binding ligands, Org. Biomol. Chem, vol.9, pp.6154-6162, 2011.
URL : https://hal.archives-ouvertes.fr/mnhn-02619199

F. Kazmi, T. Hensley, C. Pope, R. S. Funk, G. J. Loewen et al., Lysosomal Sequestration (trapping) of lipophilic amine (cationic amphiphilic) drugs in immortalized human hepatocytes (Fa2N-4 Cells), Drug Metab. Dispos, vol.41, pp.897-905, 2013.

P. Boya, R. Gonzalez-polo, D. Poncet, K. Andreau, H. L. Vieira et al., Mitochondrial membrane permeabilization is a critical step of lysosome-initiated apoptosis induced by hydroxychloroquine, Oncogene, vol.22, pp.3927-3936, 2003.

S. Aits, J. Kricker, B. Liu, A. Ellegaard, S. Hämälistö et al., Sensitive detection of lysosomal membrane permeabilization by lysosomal galectin puncta assay, Autophagy, vol.11, pp.1408-1424, 2015.

C. Ambrogio, G. Gómez-lópez, M. Falcone, A. Vidal, E. Nadal et al., Combined inhibition of DDR1 and Notch signaling is a therapeutic strategy for KRAS-driven lung adenocarcinoma, Nat. Med, vol.22, pp.270-277, 2016.

Q. Cao, X. Z. Zhong, Y. Zou, R. Murrell-lagnado, M. X. Zhu et al., Calcium release through P2X4 activates calmodulin to promote endolysosomal membrane fusion, J. Cell Biol, vol.209, pp.879-894, 2015.

Q. Cao, Y. Yang, X. Z. Zhong, and X. Dong, The lysosomal Ca 2+ release channel TRPML1 regulates lysosome size by activating calmodulin, J. Biol. Chem, vol.292, pp.8424-8435, 2017.

J. A. Martina, Y. Chen, M. Gucek, and R. Puertollano, MTORC1 functions as a transcriptional regulator of autophagy by preventing nuclear transport of TFEB, Autophagy, vol.8, pp.903-914, 2012.

C. Settembre, C. Di-malta, V. A. Polito, M. Garcia-arencibia, F. Vetrini et al., TFEB links autophagy to lysosomal biogenesis, vol.332, pp.1429-1433, 2011.

J. Beauvarlet, J. Mergny, and M. Djavaheri-mergny, Activation of the Ataxia Telangiectasia Mutated/Autophagy pathway by a G-quadruplex ligand links senescence with apoptosis, Mol. Cell Oncol, vol.6, 2019.

P. Boya, F. Reggiori, and P. Codogno, Emerging regulation and functions of autophagy, Nat. Cell Biol, vol.15, pp.713-720, 2013.

P. Boya and G. Kroemer, Lysosomal membrane permeabilization in cell death, Oncogene, vol.27, pp.6434-6451, 2008.

C. Kang, W. Huang, C. Kouh, Z. Wang, C. Cho et al., Chemical principles for the design of a novel fluorescent probe with high cancer-targeting selectivity and sensitivity, Integr. Biol, vol.5, pp.1217-1228, 2013.

A. Shivalingam, M. A. Izquierdo, A. Le-marois, A. Vy?niauskas, K. Suhling et al., The interactions between a small molecule and G-quadruplexes are visualized by fluorescence lifetime imaging microscopy, Nat. Commun, vol.6, 2015.

D. Drygin, A. Siddiqui-jain, S. O'brien, M. Schwaebe, A. Lin et al., Anticancer activity of CX-3543: A direct inhibitor of rRNA biogenesis, Cancer Res, vol.69, pp.7653-7661, 2009.

M. Faudale, S. Cogoi, and L. E. Xodo, Photoactivated cationic alkyl-substituted porphyrin binding to G4-RNA in the 5'-UTR of KRAS oncogene represses translation, Chem. Commun, vol.48, pp.874-876, 2012.

C. Montespan, S. A. Marvin, S. Austin, A. M. Burrage, B. Roger et al., Multi-layered control of Galectin-8 mediated autophagy during adenovirus cell entry through a conserved PPxY motif in the viral capsid, PLoS Pathog, vol.13, 2017.

N. M. Smith, C. Raston, C. B. Smith, and A. N. Sobolev, PEG mediated synthesis of amino-functionalised 2,4,6-triarylpyridines, Green Chem, vol.9, pp.1185-1190, 2007.

S. Zahmatkesh, Ionic liquid catalyzed synthesis and characterization of heterocyclic and optically active poly (amide-imide)s incorporating L-amino acids, Amino Acids, vol.40, pp.533-542, 2011.

M. Weiss, Acetic acid-Ammonium acetate reactions. An improved chichibabin pyridine synthesis, J. Am. Chem. Soc, vol.74, pp.200-202, 1952.