PMID- 34740311 OWN - NLM STAT- MEDLINE DCOM- 20220420 LR - 20220716 IS - 1554-8635 (Electronic) IS - 1554-8627 (Print) IS - 1554-8627 (Linking) VI - 18 IP - 3 DP - 2022 Mar TI - GNS561, a clinical-stage PPT1 inhibitor, is efficient against hepatocellular carcinoma via modulation of lysosomal functions. PG - 678-694 LID - 10.1080/15548627.2021.1988357 [doi] AB - Hepatocellular carcinoma is the most frequent primary liver cancer. Macroautophagy/autophagy inhibitors have been extensively studied in cancer but, to date, none has reached efficacy in clinical trials. In this study, we demonstrated that GNS561, a new autophagy inhibitor, whose anticancer activity was previously linked to lysosomal cell death, displayed high liver tropism and potent antitumor activity against a panel of human cancer cell lines and in two hepatocellular carcinoma in vivo models. We showed that due to its lysosomotropic properties, GNS561 could reach and specifically inhibited its enzyme target, PPT1 (palmitoyl-protein thioesterase 1), resulting in lysosomal unbound Zn(2+) accumulation, impairment of cathepsin activity, blockage of autophagic flux, altered location of MTOR (mechanistic target of rapamycin kinase), lysosomal membrane permeabilization, caspase activation and cell death. Accordingly, GNS561, for which a global phase 1b clinical trial in liver cancers was just successfully achieved, represents a promising new drug candidate and a hopeful therapeutic strategy in cancer treatment.Abbreviations: ANXA5:annexin A5; ATCC: American type culture collection; BafA1: bafilomycin A(1); BSA: bovine serum albumin; CASP3: caspase 3; CASP7: caspase 7; CASP8: caspase 8; CCND1: cyclin D1; CTSB: cathepsin B; CTSD: cathepsin D; CTSL: cathepsin L; CQ: chloroquine; iCCA: intrahepatic cholangiocarcinoma; DEN: diethylnitrosamine; DMEM: Dulbelcco's modified Eagle medium; FBS: fetal bovine serum; FITC: fluorescein isothiocyanate; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; HCC: hepatocellular carcinoma; HCQ: hydroxychloroquine; HDSF: hexadecylsulfonylfluoride; IC(50): mean half-maximal inhibitory concentration; LAMP: lysosomal associated membrane protein; LC3-II: phosphatidylethanolamine-conjugated form of MAP1LC3; LMP: lysosomal membrane permeabilization; MALDI: matrix assisted laser desorption ionization; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MKI67: marker of proliferation Ki-67; MTOR: mechanistic target of rapamycin kinase; MRI: magnetic resonance imaging; NH(4)Cl: ammonium chloride; NtBuHA: N-tert-butylhydroxylamine; PARP: poly(ADP-ribose) polymerase; PBS: phosphate-buffered saline; PPT1: palmitoyl-protein thioesterase 1; SD: standard deviation; SEM: standard error mean; vs, versus; Zn(2+): zinc ion; Z-Phe: Z-Phe-Tyt(tBu)-diazomethylketone; Z-VAD-FMK: carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]- fluoromethylketone. FAU - Brun, Sonia AU - Brun S AUID- ORCID: 0000-0001-8981-8957 AD - Genoscience Pharma, Marseille, France. FAU - Bestion, Eloine AU - Bestion E AUID- ORCID: 0000-0001-5870-3428 AD - Genoscience Pharma, Marseille, France. AD - Aix-Marseille Univ, MEPHI, APHM, IRD, IHU Mediterranee Infection, Marseille, France. FAU - Raymond, Eric AU - Raymond E AUID- ORCID: 0000-0001-5491-2708 AD - Genoscience Pharma, Marseille, France. AD - Medical Oncology, Paris Saint-Joseph Hospital, Paris, France. FAU - Bassissi, Firas AU - Bassissi F AD - Genoscience Pharma, Marseille, France. FAU - Jilkova, Zuzana Macek AU - Jilkova ZM AUID- ORCID: 0000-0002-2553-5971 AD - Institute for Advanced Biosciences, Research Center UGA/Inserm U 1209/CNRS 5309, La Tronche, France. AD - University of Grenoble Alpes, Faculte De Medecine, France. AD - Clinique Universitaire d'Hepato-gastroenterologie, Pole Digidune, Chu Grenoble, France. FAU - Mezouar, Soraya AU - Mezouar S AUID- ORCID: 0000-0002-2285-7051 AD - Genoscience Pharma, Marseille, France. FAU - Rachid, Madani AU - Rachid M AD - Genoscience Pharma, Marseille, France. FAU - Novello, Marie AU - Novello M AUID- ORCID: 0000-0003-3250-1072 AD - Genoscience Pharma, Marseille, France. FAU - Tracz, Jennifer AU - Tracz J AD - Genoscience Pharma, Marseille, France. FAU - Hamai, Ahmed AU - Hamai A AD - Institut Necker-Enfants Malades, Inserm U1151-CNRS UMR, Paris, France. AD - University of Paris Descartes-Sorbonne Paris Cite, Paris, France. FAU - Lalmanach, Gilles AU - Lalmanach G AUID- ORCID: 0000-0001-8562-4821 AD - Inserm, UMR1100, Centre d'Etude Des Pathologies Respiratoires, Equipe "Mecanismes Proteolytiques Dans l'Inflammation", Tours, France. AD - University of Tours, Tours, France. FAU - Vanderlynden, Lise AU - Vanderlynden L AD - Inserm, UMR1100, Centre d'Etude Des Pathologies Respiratoires, Equipe "Mecanismes Proteolytiques Dans l'Inflammation", Tours, France. AD - University of Tours, Tours, France. FAU - Legouffe, Raphael AU - Legouffe R AD - ImaBiotech, Loos, France. FAU - Stauber, Jonathan AU - Stauber J AD - ImaBiotech, Billerica, USA. FAU - Schubert, Thomas AU - Schubert T AD - 2Bind GmbH, Regensburg, Germany. FAU - Plach, Maximilian G AU - Plach MG AD - 2Bind GmbH, Regensburg, Germany. FAU - Courcambeck, Jerome AU - Courcambeck J AD - Genoscience Pharma, Marseille, France. FAU - Drouot, Cyrille AU - Drouot C AD - Genoscience Pharma, Marseille, France. FAU - Jacquemot, Guillaume AU - Jacquemot G AD - Genoscience Pharma, Marseille, France. FAU - Serdjebi, Cindy AU - Serdjebi C AD - Genoscience Pharma, Marseille, France. FAU - Roth, Gael AU - Roth G AUID- ORCID: 0000-0001-5822-4320 AD - Institute for Advanced Biosciences, Research Center UGA/Inserm U 1209/CNRS 5309, La Tronche, France. AD - University of Grenoble Alpes, Faculte De Medecine, France. AD - Clinique Universitaire d'Hepato-gastroenterologie, Pole Digidune, Chu Grenoble, France. FAU - Baudoin, Jean-Pierre AU - Baudoin JP AUID- ORCID: 0000-0001-7361-5653 AD - Aix-Marseille Univ, MEPHI, APHM, IRD, IHU Mediterranee Infection, Marseille, France. FAU - Ansaldi, Christelle AU - Ansaldi C AD - Genoscience Pharma, Marseille, France. FAU - Decaens, Thomas AU - Decaens T AUID- ORCID: 0000-0003-0928-0048 AD - Institute for Advanced Biosciences, Research Center UGA/Inserm U 1209/CNRS 5309, La Tronche, France. AD - University of Grenoble Alpes, Faculte De Medecine, France. AD - Clinique Universitaire d'Hepato-gastroenterologie, Pole Digidune, Chu Grenoble, France. FAU - Halfon, Philippe AU - Halfon P AD - Genoscience Pharma, Marseille, France. LA - eng PT - Journal Article DEP - 20211105 PL - United States TA - Autophagy JT - Autophagy JID - 101265188 RN - 0 (Antineoplastic Agents) RN - 0 (Membrane Proteins) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) RN - EC 3.1.2.- (Thiolester Hydrolases) RN - EC 3.1.2.22 (PPT1 protein, human) RN - W36ZG6FT64 (Sirolimus) SB - IM MH - *Antineoplastic Agents/pharmacology MH - Autophagosomes/metabolism MH - Autophagy/physiology MH - *Carcinoma, Hepatocellular/drug therapy/metabolism MH - Humans MH - *Liver Neoplasms/drug therapy/metabolism MH - Lysosomes/metabolism MH - Membrane Proteins/metabolism MH - Sirolimus/pharmacology MH - TOR Serine-Threonine Kinases/metabolism MH - Thiolester Hydrolases/metabolism/pharmacology PMC - PMC9037544 OTO - NOTNLM OT - Antitumor OT - PPT1 OT - autophagy OT - liver cancer OT - lysosome OT - mtor COIS- SB, EB, FB, ER, SM, MR, MN, JT, JC, CD, GJ, CS, CA and PH are employees of Genoscience Pharma. SB, ER, FB, CD, CS, CA and PH are shareholders of Genoscience Pharma. SB, FB, JC and PH are co-inventors of a pending patent. The other authors declare that they have no conflicts of interest to report. EDAT- 2021/11/07 06:00 MHDA- 2022/04/21 06:00 PMCR- 2021/11/05 CRDT- 2021/11/06 05:19 PHST- 2021/11/07 06:00 [pubmed] PHST- 2022/04/21 06:00 [medline] PHST- 2021/11/06 05:19 [entrez] PHST- 2021/11/05 00:00 [pmc-release] AID - 1988357 [pii] AID - 10.1080/15548627.2021.1988357 [doi] PST - ppublish SO - Autophagy. 2022 Mar;18(3):678-694. doi: 10.1080/15548627.2021.1988357. Epub 2021 Nov 5.