Search

Your search keyword '"Allan Sauvat"' showing total 81 results
Start Over Author "Allan Sauvat"
81 results on '"Allan Sauvat"'

1. Inhibition of acyl-CoA binding protein (ACBP) by means of a GABAARγ2-derived peptide

Author

Gerasimos Anagnostopoulos, Ester Saavedra, Flavia Lambertucci, Omar Motiño, Jordan Dimitrov, David Roiz-Valle, Victor Quesada, Karla Alvarez-Valadez, Hui Chen, Allan Sauvat, Yan Rong, Uxía Nogueira-Recalde, Sijing Li, Léa Montégut, Mojgan Djavaheri-Mergny, Maria Castedo, Carlos Lopez-Otin, Maria Chiara Maiuri, Isabelle Martins, and Guido Kroemer

Subjects
Cytology, QH573-671
Abstract

Abstract Acyl-CoA binding protein (ACBP) encoded by diazepam binding inhibitor (DBI) is an extracellular inhibitor of autophagy acting on the gamma-aminobutyric acid A receptor (GABAAR) γ2 subunit (GABAARγ2). Here, we show that lipoanabolic diets cause an upregulation of GABAARγ2 protein in liver hepatocytes but not in other major organs. ACBP/DBI inhibition by systemically injected antibodies has been demonstrated to mediate anorexigenic and organ-protective, autophagy-dependent effects. Here, we set out to develop a new strategy for developing ACBP/DBI antagonists. For this, we built a molecular model of the interaction of ACBP/DBI with peptides derived from GABAARγ2. We then validated the interaction between recombinant and native ACBP/DBI protein and a GABAARγ2-derived eicosapeptide (but not its F77I mutant) by pull down experiments or surface plasmon resonance. The GABAARγ2-derived eicosapeptide inhibited the metabolic activation of hepatocytes by recombinant ACBP/DBI protein in vitro. Moreover, the GABAARγ2-derived eicosapeptide (but not its F77I-mutated control) blocked appetite stimulation by recombinant ACBP/DBI in vivo, induced autophagy in the liver, and protected mice against the hepatotoxin concanavalin A. We conclude that peptidomimetics disrupting the interaction between ACBP/DBI and GABAARγ2 might be used as ACBP/DBI antagonists. This strategy might lead to the future development of clinically relevant small molecules of the ACBP/DBI system.

Published
2024
Full Text
View/download PDF

2. 3,4-dimethoxychalcone induces autophagy and reduces neointimal hyperplasia and aortic lesions in mouse models of atherosclerosis

Author

Giulia Cerrato, Carlota Alvarez-Lucena, Allan Sauvat, Yanhua Hu, Sabrina Forveille, Guo Chen, Sylvère Durand, Fanny Aprahamian, Marion Leduc, Omar Motiño, Lisardo Boscá, Qingbo Xu, Oliver Kepp, and Guido Kroemer

Subjects
Cytology, QH573-671
Abstract

Abstract Autophagy inducers can prevent cardiovascular aging and age-associated diseases including atherosclerosis. Therefore, we hypothesized that autophagy-inducing compounds that act on atherosclerosis-relevant cells might have a protective role in the development of atherosclerosis. Here we identified 3,4-dimethoxychalcone (3,4-DC) as an inducer of autophagy in several cell lines from endothelial, myocardial and myeloid/macrophagic origin, as demonstrated by the aggregation of the autophagosome marker GFP-LC3 in the cytoplasm of cells, as well as the downregulation of its nuclear pool indicative of autophagic flux. In this respect, 3,4-DC showed a broader autophagy-inducing activity than another chalcone (4,4- dimethoxychalcone), spermidine and triethylene tetramine. Thus, we characterized the potential antiatherogenic activity of 3,4-DC in two different mouse models, namely, (i) neointima formation with smooth muscle expansion of vein segments grafted to the carotid artery and (ii) genetically predisposed ApoE −/− mice fed an atherogenic diet. In the vein graft model, local application of 3,4-DC was able to maintain the lumen of vessels and to reduce neointima lesions. In the diet-induced model, intraperitoneal injections of 3,4-DC significantly reduced the number of atherosclerotic lesions in the aorta. In conclusion, 3,4-DC stands out as an autophagy inducer with potent antiatherogenic activity.

Published
2023
Full Text
View/download PDF

3. Formyl peptide receptor-1 (FPR1) represses intestinal oncogenesis

Author

Julie Le Naour, Léa Montégut, Yuhong Pan, Sarah Adriana Scuderi, Pierre Cordier, Adrien Joseph, Allan Sauvat, Valerio Iebba, Juliette Paillet, Gladys Ferrere, Ludivine Brechard, Claire Mulot, Grégory Dubourg, Laurence Zitvogel, Jonathan G. Pol, Erika Vacchelli, Pierre-Laurent Puig, and Guido Kroemer

Subjects
Bone marrow-derived dendritic cells, chemotaxis, immunogenic chemotherapy, immunosurveillance, tumor microenvironment, Immunologic diseases. Allergy, RC581-607, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282
Abstract

ABSTRACTFormyl peptide receptor-1 (FPR1) is a pattern recognition receptor that is mostly expressed by myeloid cells. In patients with colorectal cancer (CRC), a loss-of-function polymorphism (rs867228) in the gene coding for FPR1 has been associated with reduced responses to chemotherapy or chemoradiotherapy. Moreover, rs867228 is associated with accelerated esophageal and colorectal carcinogenesis. Here, we show that dendritic cells from Fpr1−/− mice exhibit reduced migration in response to chemotherapy-treated CRC cells. Moreover, Fpr1−/− mice are particularly susceptible to chronic ulcerative colitis and colorectal oncogenesis induced by the mutagen azoxymethane followed by oral dextran sodium sulfate, a detergent that induces colitis. These experiments were performed after initial co-housing of Fpr1−/− mice and wild-type controls, precluding major Fpr1-driven differences in the microbiota. Pharmacological inhibition of Fpr1 by cyclosporin H also tended to increase intestinal oncogenesis in mice bearing the ApcMin mutation, and this effect was reversed by the anti-inflammatory drug sulindac. We conclude that defective FPR1 signaling favors intestinal tumorigenesis through the modulation of the innate inflammatory/immune response.

Published
2023
Full Text
View/download PDF

4. Correction: Metabolomic analyses of COVID-19 patients unravel stage-dependent and prognostic biomarkers

Author

François-Xavier Danlos, Claudia Grajeda-Iglesias, Sylvère Durand, Allan Sauvat, Mathilde Roumier, Delphine Cantin, Emeline Colomba, Julien Rohmer, Fanny Pommeret, Giulia Baciarello, Christophe Willekens, Marc Vasse, Frank Griscelli, Jean-Eudes Fahrner, Anne-Gaëlle Goubet, Agathe Dubuisson, Lisa Derosa, Nitharsshini Nirmalathasan, Delphine Bredel, Séverine Mouraud, Caroline Pradon, Annabelle Stoclin, Flore Rozenberg, Jérôme Duchemin, Georges Jourdi, Syrine Ellouze, Françoise Levavasseur, Laurence Albigès, Jean-Charles Soria, Fabrice Barlesi, Eric Solary, Fabrice André, Frédéric Pène, Félix Ackerman, Luc Mouthon, Laurence Zitvogel, Aurélien Marabelle, Jean-Marie Michot, Michaela Fontenay, and Guido Kroemer

Subjects
Cytology, QH573-671
Published
2024
Full Text
View/download PDF

5. Everolimus and plicamycin specifically target chemoresistant colorectal cancer cells of the CMS4 subtype

Author

Jiayin Deng, Ai-Ling Tian, Hui Pan, Allan Sauvat, Marion Leduc, Peng Liu, Liwei Zhao, Shuai Zhang, Hui Chen, Valérie Taly, Pierre Laurent-Puig, Laura Senovilla, Yingqiu Li, Guido Kroemer, and Oliver Kepp

Subjects
Cytology, QH573-671
Abstract

Abstract Colorectal cancers (CRC) can be classified into four consensus molecular subtypes (CMS), among which CMS1 has the best prognosis, contrasting with CMS4 that has the worst outcome. CMS4 CRC is notoriously resistant against therapeutic interventions, as demonstrated by preclinical studies and retrospective clinical observations. Here, we report the finding that two clinically employed agents, everolimus (EVE) and plicamycin (PLI), efficiently target the prototypic CMS4 cell line MDST8. As compared to the prototypic CMS1 cell line LoVo, MDST8 cells treated with EVE or PLI demonstrated stronger cytostatic and cytotoxic effects, increased signs of apoptosis and autophagy, as well as a more pronounced inhibition of DNA-to-RNA transcription and RNA-to-protein translation. Moreover, nontoxic doses of EVE and PLI induced the shrinkage of MDST8 tumors in mice, yet had only minor tumor growth-reducing effects on LoVo tumors. Altogether, these results suggest that EVE and PLI should be evaluated for their clinical activity against CMS4 CRC.

Published
2021
Full Text
View/download PDF

6. Antibody–drug conjugates harboring a kinesin spindle protein inhibitor with immunostimulatory properties

Author

Anette Sommer, Sandra Berndt, Hans-Georg Lerchen, Sabrina Forveille, Allan Sauvat, Dominik Mumberg, Guido Kroemer, and Oliver Kepp

Subjects
immunogenic cell death, calreticulin, type i interferon response, adaptive immunity, ksp inhibitors, adcs, Immunologic diseases. Allergy, RC581-607, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282
Abstract

Antibody–drug conjugates (ADCs) are used to target cancer cells by means of antibodies directed to tumor-associated antigens, causing the incorporation of a cytotoxic payload into target cells. Here, we characterized the mode of action of ADC costing of a TWEAKR-specific monoclonal antibody conjugated to a small molecule kinesin spindle protein (KSP) inhibitor (KSPi). These TWEAKR-KSPi-ADCs showed strong efficacy in a TWEAKR expressing CT26 colon cancer model in mice. TWEAKR-KSPi-ADCs controlled the growth of CT26 colon cancers in immunodeficient as well as in immunocompetent mice. However, when treated with suboptimal doses, TWEAKR-KSPi-ADCs were still active in immunocompetent but not in immunodeficient mice, indicating that TWEAKR-KSPi-ADCs act – in addition to the cytotoxic mode of action – through an immunological mechanism. Indeed, in vitro experiments performed with a cell-permeable small molecule KSPi closely related to the active payload released from the TWEAKR-KSPi-ADCs revealed that KSPi was capable of stimulating several hallmarks of immunogenic cell death (ICD) on three different human cancer cell lines: cellular release of adenosine triphosphate (ATP) and high mobility group B1 protein (HMGB1), exposure of calreticulin on the cell surface as well as a transcriptional type-I interferon response. Further, in vivo experiments confirmed that treatment with TWEAKR-KSPi-ADCs activated immune responses via enhancing the infiltration of CD4+ and CD8+ T lymphocytes in tumors and the local production of interferon-γ, interleukin-2, and tumor necrosis factor-α. In conclusion, the antineoplastic effects of TWEAKR-KSPi-ADCs can partly be attributed to its ICD-stimulatory properties.

Published
2022
Full Text
View/download PDF

7. Metabolomic analyses of COVID-19 patients unravel stage-dependent and prognostic biomarkers

Author

François-Xavier Danlos, Claudia Grajeda-Iglesias, Sylvère Durand, Allan Sauvat, Mathilde Roumier, Delphine Cantin, Emeline Colomba, Julien Rohmer, Fanny Pommeret, Giulia Baciarello, Christophe Willekens, Marc Vasse, Frank Griscelli, Jean-Eudes Fahrner, Anne-Gaëlle Goubet, Agathe Dubuisson, Lisa Derosa, Nitharsshini Nirmalathasan, Delphine Bredel, Séverine Mouraud, Caroline Pradon, Annabelle Stoclin, Flore Rozenberg, Jérôme Duchemin, Georges Jourdi, Syrine Ellouze, Françoise Levavasseur, Laurence Albigès, Jean-Charles Soria, Fabrice Barlesi, Eric Solary, Fabrice André, Frédéric Pène, Félix Ackerman, Luc Mouthon, Laurence Zitvogel, Aurélien Marabelle, Jean-Marie Michot, Michaela Fontenay, and Guido Kroemer

Subjects
Cytology, QH573-671
Abstract

Abstract The circulating metabolome provides a snapshot of the physiological state of the organism responding to pathogenic challenges. Here we report alterations in the plasma metabolome reflecting the clinical presentation of COVID-19 patients with mild (ambulatory) diseases, moderate disease (radiologically confirmed pneumonitis, hospitalization and oxygen therapy), and critical disease (in intensive care). This analysis revealed major disease- and stage-associated shifts in the metabolome, meaning that at least 77 metabolites including amino acids, lipids, polyamines and sugars, as well as their derivatives, were altered in critical COVID-19 patient’s plasma as compared to mild COVID-19 patients. Among a uniformly moderate cohort of patients who received tocilizumab, only 10 metabolites were different among individuals with a favorable evolution as compared to those who required transfer into the intensive care unit. The elevation of one single metabolite, anthranilic acid, had a poor prognostic value, correlating with the maintenance of high interleukin-10 and -18 levels. Given that products of the kynurenine pathway including anthranilic acid have immunosuppressive properties, we speculate on the therapeutic utility to inhibit the rate-limiting enzymes of this pathway including indoleamine 2,3-dioxygenase and tryptophan 2,3-dioxygenase.

Published
2021
Full Text
View/download PDF

8. Cancer cell-autonomous overactivation of PARP1 compromises immunosurveillance in non-small cell lung cancer

Author

Adrien Joseph, Guido Kroemer, Antoine Lafarge, Oliver Kepp, Jonathan Pol, Sylvere Durand, Marion Leduc, Pan Juncheng, Florine Obrist, Judith Michels, Maria Castedo, Allan Sauvat, Giulia Cerrato, Isabelle Martins, Ester Saavedra, Sijing Li, Sarah Lévesque, Fanny Aprahamian, and Nitharsshini Nirmalathansan

Subjects
Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282
Published
2022
Full Text
View/download PDF

9. Local anesthetics elicit immune-dependent anticancer effects

Author

Shuai Zhang, Peng Liu, Wei Xie, Fabrice Barlesi, Guido Kroemer, Oliver Kepp, Juliette Humeau, Jonathan Pol, Liwei Zhao, Lucillia Bezu, Alejandra Wu Chuang, Allan Sauvat, Giulia Cerrato, Julie Le Naour, and Peter van Endert

Subjects
Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282
Published
2022
Full Text
View/download PDF

10. Autophagy-mediated metabolic effects of aspirin

Author

Francesca Castoldi, Juliette Humeau, Isabelle Martins, Sylvie Lachkar, Damarys Loew, Florent Dingli, Sylvère Durand, David Enot, Noëlie Bossut, Alexis Chery, Fanny Aprahamian, Yohann Demont, Paule Opolon, Nicolas Signolle, Allan Sauvat, Michaela Semeraro, Lucillia Bezu, Elisa Elena Baracco, Erika Vacchelli, Jonathan G. Pol, Sarah Lévesque, Norma Bloy, Valentina Sica, Maria Chiara Maiuri, Guido Kroemer, and Federico Pietrocola

Subjects
Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282, Cytology, QH573-671
Abstract

Abstract Salicylate, the active derivative of aspirin (acetylsalicylate), recapitulates the mode of action of caloric restriction inasmuch as it stimulates autophagy through the inhibition of the acetyltransferase activity of EP300. Here, we directly compared the metabolic effects of aspirin medication with those elicited by 48 h fasting in mice, revealing convergent alterations in the plasma and the heart metabolome. Aspirin caused a transient reduction of general protein acetylation in blood leukocytes, accompanied by the induction of autophagy. However, these effects on global protein acetylation could not be attributed to the mere inhibition of EP300, as determined by epistatic experiments and exploration of the acetyl-proteome from salicylate-treated EP300-deficient cells. Aspirin reduced high-fat diet-induced obesity, diabetes, and hepatosteatosis. These aspirin effects were observed in autophagy-competent mice but not in two different models of genetic (Atg4b −/− or Bcln1 +/−) autophagy-deficiency. Aspirin also improved tumor control by immunogenic chemotherapeutics, and this effect was lost in T cell-deficient mice, as well as upon knockdown of an essential autophagy gene (Atg5) in cancer cells. Hence, the health-improving effects of aspirin depend on autophagy.

Published
2020
Full Text
View/download PDF

11. Inhibition of transcription by dactinomycin reveals a new characteristic of immunogenic cell stress

Author

Juliette Humeau, Allan Sauvat, Giulia Cerrato, Wei Xie, Friedemann Loos, Francesca Iannantuoni, Lucillia Bezu, Sarah Lévesque, Juliette Paillet, Jonathan Pol, Marion Leduc, Laurence Zitvogel, Hugues de Thé, Oliver Kepp, and Guido Kroemer

Subjects
dactinomycin, eIF2α phosphorylation, immunogenic cell death, transcription, translation, Medicine (General), R5-920, Genetics, QH426-470
Abstract

Abstract Chemotherapy still constitutes the standard of care for the treatment of most neoplastic diseases. Certain chemotherapeutics from the oncological armamentarium are able to trigger pre‐mortem stress signals that lead to immunogenic cell death (ICD), thus inducing an antitumor immune response and mediating long‐term tumor growth reduction. Here, we used an established model, built on artificial intelligence to identify, among a library of 50,000 compounds, anticancer agents that, based on their molecular descriptors, were predicted to induce ICD. This algorithm led us to the identification of dactinomycin (DACT, best known as actinomycin D), a highly potent cytotoxicant and ICD inducer that mediates immune‐dependent anticancer effects in vivo. Since DACT is commonly used as an inhibitor of DNA to RNA transcription, we investigated whether other experimentally established or algorithm‐selected, clinically employed ICD inducers would share this characteristic. As a common leitmotif, a panel of pharmacological ICD stimulators inhibited transcription and secondarily translation. These results establish the inhibition of RNA synthesis as an initial event for ICD induction.

Published
2020
Full Text
View/download PDF

12. 3,4‐Dimethoxychalcone induces autophagy through activation of the transcription factors TFE3 and TFEB

Author

Guo Chen, Wei Xie, Jihoon Nah, Allan Sauvat, Peng Liu, Federico Pietrocola, Valentina Sica, Didac Carmona‐Gutierrez, Andreas Zimmermann, Tobias Pendl, Jelena Tadic, Martina Bergmann, Sebastian J Hofer, Lana Domuz, Sylvie Lachkar, Maria Markaki, Nektarios Tavernarakis, Junichi Sadoshima, Frank Madeo, Oliver Kepp, and Guido Kroemer

Subjects
caloric restriction, caloric restriction mimetic, cardioprotection, TFE3, TFEB, Medicine (General), R5-920, Genetics, QH426-470
Abstract

Abstract Caloric restriction mimetics (CRMs) are natural or synthetic compounds that mimic the health‐promoting and longevity‐extending effects of caloric restriction. CRMs provoke the deacetylation of cellular proteins coupled to an increase in autophagic flux in the absence of toxicity. Here, we report the identification of a novel candidate CRM, namely 3,4‐dimethoxychalcone (3,4‐DC), among a library of polyphenols. When added to several different human cell lines, 3,4‐DC induced the deacetylation of cytoplasmic proteins and stimulated autophagic flux. At difference with other well‐characterized CRMs, 3,4‐DC, however, required transcription factor EB (TFEB)‐ and E3 (TFE3)‐dependent gene transcription and mRNA translation to trigger autophagy. 3,4‐DC stimulated the translocation of TFEB and TFE3 into nuclei both in vitro and in vivo, in hepatocytes and cardiomyocytes. 3,4‐DC induced autophagy in vitro and in mouse organs, mediated autophagy‐dependent cardioprotective effects, and improved the efficacy of anticancer chemotherapy in vivo. Altogether, our results suggest that 3,4‐DC is a novel CRM with a previously unrecognized mode of action.

Published
2019
Full Text
View/download PDF

14. Trans-Fats Inhibit Autophagy Induced by Saturated Fatty Acids

Author

Allan Sauvat, Guo Chen, Kevin Müller, Mingming Tong, Fanny Aprahamian, Sylvère Durand, Giulia Cerrato, Lucillia Bezu, Marion Leduc, Joakim Franz, Patrick Rockenfeller, Junichi Sadoshima, Frank Madeo, Oliver Kepp, and Guido Kroemer

Subjects
Medicine, Medicine (General), R5-920
Abstract

Depending on the length of their carbon backbone and their saturation status, natural fatty acids have rather distinct biological effects. Thus, longevity of model organisms is increased by extra supply of the most abundant natural cis-unsaturated fatty acid, oleic acid, but not by that of the most abundant saturated fatty acid, palmitic acid. Here, we systematically compared the capacity of different saturated, cis-unsaturated and alien (industrial or ruminant) trans-unsaturated fatty acids to provoke cellular stress in vitro, on cultured human cells expressing a battery of distinct biosensors that detect signs of autophagy, Golgi stress and the unfolded protein response. In contrast to cis-unsaturated fatty acids, trans-unsaturated fatty acids failed to stimulate signs of autophagy including the formation of GFP-LC3B-positive puncta, production of phosphatidylinositol-3-phosphate, and activation of the transcription factor TFEB. When combined effects were assessed, several trans-unsaturated fatty acids including elaidic acid (the trans-isomer of oleate), linoelaidic acid, trans-vaccenic acid and palmitelaidic acid, were highly efficient in suppressing autophagy and endoplasmic reticulum stress induced by palmitic, but not by oleic acid. Elaidic acid also inhibited autophagy induction by palmitic acid in vivo, in mouse livers and hearts. We conclude that the well-established, though mechanistically enigmatic toxicity of trans-unsaturated fatty acids may reside in their capacity to abolish cytoprotective stress responses induced by saturated fatty acids. Keywords: Fasting, Ketogenic diet, Immune response, Immunosurveillance, Obesity, Aging, Cytoprotection, Systems biology

Published
2018
Full Text
View/download PDF

15. Lurbinectedin synergizes with immune checkpoint blockade to generate anticancer immunity

Author

Wei Xie, Sabrina Forveille, Kristina Iribarren, Allan Sauvat, Laura Senovilla, Yan Wang, Juliette Humeau, Maria Perez-Lanzon, Heng Zhou, Juan F. Martínez-Leal, Guido Kroemer, and Oliver Kepp

Subjects
anticancer immunity, immunogenic cell death, checkpoint blockade, tumor clearance, Immunologic diseases. Allergy, RC581-607, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282
Abstract

Systemic treatment with the active transcription inhibitor lurbinectedin aims at inducing tumor cell death in hyperproliferative neoplasms. Here we show that cell death induced by lurbinectedin reinstates and enhances systemic anticancer immune responses. Lurbinectedin treatment showed traits of immunogenic cell death, including the exposure of calreticulin, the release of ATP, the exodus of high mobility group box 1 (HMGB1) and type 1 interferon responses in vitro. Lurbinectedin treated cells induced antitumor immunity when injected into immunocompetent animals and treatment of transplanted fibrosarcomas reduced tumor growth in immunocompetent yet not in immunodeficient hosts. Anticancer effects resulting from lurbinectedin treatment were boosted in combination with PD-1 and CTLA-4 double immune checkpoint blockade (ICB), and lurbinectedin combined with double ICB exhibited strong antineoplastic effects. Cured animals exhibited long term immune memory effects that rendered them resistant to rechallenge with syngeneic tumors underlining the potency of combination therapy with lurbinectedin.

Published
2019
Full Text
View/download PDF

16. A fluorescent biosensor-based platform for the discovery of immunogenic cancer cell death inducers

Author

Oliver Kepp, Allan Sauvat, Marion Leduc, Sabrina Forveille, Peng Liu, Liwei Zhao, Lucillia Bezu, Wei Xie, Laurence Zitvogel, and Guido Kroemer

Subjects
biosensor, artificial intelligence, imaging, automatization, Immunologic diseases. Allergy, RC581-607, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282
Abstract

Systemic anticancer immunity can be reinstated via the induction of immunogenic cell death (ICD) in malignant cells. Thus, certain classes of cytotoxic compounds, for example, anthracyclines, oxaliplatin and taxanes are endowed with the capacity to act on cancer cells to ignite premortem stress pathways that lead to the surface exposure of calreticulin (CALR) and the cellular release of adenosine triphosphate, annexin A1, high mobility group B1 and type-1 interferons. Altogether, these alterations constitute the hallmarks of ICD. Here we report the design of a discovery pipeline for the identification of novel ICD inducers by means of a phenotypic screening platform. The use of fluorescent biosensors as proxies for the manifestation of ICD hallmarks has enabled the exploration of large collections of chemical compounds by automatized screening routines. Imaging-based assessment and phenotypic selection led to the identification of potential ICD inducers that could be validated further in vitro and in vivo, confirming that bona fide ICD inducers possess the capacity to induce immunological long-term memory and to confer resistance against rechallenge with syngeneic tumors. Machine learning algorithms analyzing the physicochemical properties of ICD inducers can assist in the preselection of compounds with potential ICD-stimulatory properties, further accelerating the screening efforts designed to develop new immunotherapeutic agents.

Published
2019
Full Text
View/download PDF

17. Lethal Poisoning of Cancer Cells by Respiratory Chain Inhibition plus Dimethyl α-Ketoglutarate

Author

Valentina Sica, Jose Manuel Bravo-San Pedro, Valentina Izzo, Jonathan Pol, Sandra Pierredon, David Enot, Sylvère Durand, Noélie Bossut, Alexis Chery, Sylvie Souquere, Gerard Pierron, Evangelia Vartholomaiou, Naoufal Zamzami, Thierry Soussi, Allan Sauvat, Laura Mondragón, Oliver Kepp, Lorenzo Galluzzi, Jean-Claude Martinou, Holger Hess-Stumpp, Karl Ziegelbauer, Guido Kroemer, and Maria Chiara Maiuri

Subjects
Biology (General), QH301-705.5
Abstract

Summary: Inhibition of oxidative phosphorylation (OXPHOS) by 1-cyclopropyl-4-(4-[(5-methyl-3-(3-[4-(trifluoromethoxy)phenyl]-1,2,4-oxadiazol-5-yl)-1H-pyrazol-1-yl)methyl]pyridin-2-yl)piperazine (BAY87-2243, abbreviated as B87), a complex I inhibitor, fails to kill human cancer cells in vitro. Driven by this consideration, we attempted to identify agents that engage in synthetically lethal interactions with B87. Here, we report that dimethyl α-ketoglutarate (DMKG), a cell-permeable precursor of α-ketoglutarate that lacks toxicity on its own, kills cancer cells when combined with B87 or other inhibitors of OXPHOS. DMKG improved the antineoplastic effect of B87, both in vitro and in vivo. This combination caused MDM2-dependent, tumor suppressor protein p53 (TP53)-independent transcriptional reprogramming and alternative exon usage affecting multiple glycolytic enzymes, completely blocking glycolysis. Simultaneous inhibition of OXPHOS and glycolysis provoked a bioenergetic catastrophe culminating in the activation of a cell death program that involved disruption of the mitochondrial network and activation of PARP1, AIFM1, and APEX1. These results unveil a metabolic liability of human cancer cells that may be harnessed for the development of therapeutic regimens. : Sica et al. show that respiratory chain inhibition by 1-cyclopropyl-4-(4-[(5-methyl-3-(3-[4-(trifluoromethoxy)phenyl]-1,2,4-oxadiazol-5-yl)-1H-pyrazol-1-yl)methyl]pyridin-2-yl)piperazine (BAY87-2243, abbreviated as B87) becomes lethal for cancer cells when glycolysis is simultaneously suppressed. When combined with B87, dimethyl α-ketoglutarate acquires the capacity to suppress glycolysis, thus lethally poisoning bioenergetics metabolism. This therapeutic combination effect relies on transcriptional reprogramming that can be reverted by pharmacological inhibition of MDM2. Keywords: MDM2, Krebs cycle, glycolysis, mitochondrial fragmentation, regulated cell death, parthanatos, cancer metabolism

Published
2019
Full Text
View/download PDF

18. eIF2α phosphorylation: A hallmark of immunogenic cell death

Author

Lucillia Bezu, Allan Sauvat, Juliette Humeau, Marion Leduc, Oliver Kepp, and Guido Kroemer

Subjects
anthracyclines, cancer, chemotherapy, er stress, unfolded protein response, Immunologic diseases. Allergy, RC581-607, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282
Abstract

Immunogenic cell death (ICD) induced by anticancer chemotherapeutics is usually preceded by premortem stress affecting the endoplasmic reticulum (ER). This ER stress does not reflect a canonical unfolded protein response (UPR) but rather manifests solely at the level of the phosphorylation of eIF2α. eIF2α phosphorylation is hence a quintessential hallmark of ICD that can be detected by immunohistochemistry in tumor samples.

Published
2018
Full Text
View/download PDF

22. Supplementary Figure from Escherichia coli–Specific CXCL13-Producing TFH Are Associated with Clinical Efficacy of Neoadjuvant PD-1 Blockade against Muscle-Invasive Bladder Cancer

Author

Yohann Loriot, Laurence Zitvogel, Jean-Yves Scoazec, Guido Kroemer, Miriam Merad, Fabrice Andre, Benjamin Besse, Michiel S. Van Der Heijden, Nick Van Dijk, Jeroen Van Dorp, Aurélien Marabelle, Lisa Derosa, Gwénaël Le Teuff, Florent Ginhoux, Mathieu Rouanne, Jacques Fieschi, Yves Allory, Camelia Radulescu, Etienne Rouleau, Romain Daillere, Adeline Mallet, Maryse Moya-Nilges, Nadège Cayet, Ivo Gomperts Boneca, Shaima Belhechmi, Baptiste Archambaud, Thibault Raoult, Aymeric Silvin, Kevin Mulder, Garett Dunsmore, François-Xavier Danlos, Camille Bleriot, Jacques Bou Khalil, Gabriel Haddad, Caroline Davin, Mounia Filahi, Thomas Sbarrato, Allan Sauvat, Nicolas Signolle, Virginie Marty, Pierre Ly, Caroline Flament, Marine Mazzenga, Agathe Dubuisson, Idir Ouzaid, Evanguelos Xylinas, Morgan Roupret, Luca Campedel, Carole Helissey, Gwenaelle Gravis, Géraldine Pignot, François Audenet, Constance Thibault, Cédric Lebacle, Cassandra Thelemaque, Maxime Descartes Mbogning-Fonkou, Marianne Gazzano, Isabelle Peguillet, Carolina Alves Costa Silva, Leonardo Lordello, and Anne-Gaëlle Goubet

Abstract

Supplementary Figure from Escherichia coli–Specific CXCL13-Producing TFH Are Associated with Clinical Efficacy of Neoadjuvant PD-1 Blockade against Muscle-Invasive Bladder Cancer

Published
2023
Full Text
View/download PDF

23. Data from Escherichia coli–Specific CXCL13-Producing TFH Are Associated with Clinical Efficacy of Neoadjuvant PD-1 Blockade against Muscle-Invasive Bladder Cancer

Author

Yohann Loriot, Laurence Zitvogel, Jean-Yves Scoazec, Guido Kroemer, Miriam Merad, Fabrice Andre, Benjamin Besse, Michiel S. Van Der Heijden, Nick Van Dijk, Jeroen Van Dorp, Aurélien Marabelle, Lisa Derosa, Gwénaël Le Teuff, Florent Ginhoux, Mathieu Rouanne, Jacques Fieschi, Yves Allory, Camelia Radulescu, Etienne Rouleau, Romain Daillere, Adeline Mallet, Maryse Moya-Nilges, Nadège Cayet, Ivo Gomperts Boneca, Shaima Belhechmi, Baptiste Archambaud, Thibault Raoult, Aymeric Silvin, Kevin Mulder, Garett Dunsmore, François-Xavier Danlos, Camille Bleriot, Jacques Bou Khalil, Gabriel Haddad, Caroline Davin, Mounia Filahi, Thomas Sbarrato, Allan Sauvat, Nicolas Signolle, Virginie Marty, Pierre Ly, Caroline Flament, Marine Mazzenga, Agathe Dubuisson, Idir Ouzaid, Evanguelos Xylinas, Morgan Roupret, Luca Campedel, Carole Helissey, Gwenaelle Gravis, Géraldine Pignot, François Audenet, Constance Thibault, Cédric Lebacle, Cassandra Thelemaque, Maxime Descartes Mbogning-Fonkou, Marianne Gazzano, Isabelle Peguillet, Carolina Alves Costa Silva, Leonardo Lordello, and Anne-Gaëlle Goubet

Abstract

Biomarkers guiding the neoadjuvant use of immune-checkpoint blockers (ICB) are needed for patients with localized muscle-invasive bladder cancers (MIBC). Profiling tumor and blood samples, we found that follicular helper CD4+ T cells (TFH) are among the best therapeutic targets of pembrolizumab correlating with progression-free survival. TFH were associated with tumoral CD8 and PD-L1 expression at baseline and the induction of tertiary lymphoid structures after pembrolizumab. Blood central memory TFH accumulated in tumors where they produce CXCL13, a chemokine found in the plasma of responders only. IgG4+CD38+ TFH residing in bladder tissues correlated with clinical benefit. Finally, TFH and IgG directed against urothelium-invasive Escherichia coli dictated clinical responses to pembrolizumab in three independent cohorts. The links between tumor infection and success of ICB immunomodulation should be prospectively assessed at a larger scale.Significance:In patients with bladder cancer treated with neoadjuvant pembrolizumab, E. coli–specific CXCL13 producing TFH and IgG constitute biomarkers that predict clinical benefit. Beyond its role as a biomarker, such immune responses against E. coli might be harnessed for future therapeutic strategies.This article is highlighted in the In This Issue feature, p. 2221

Published
2023
Full Text
View/download PDF

24. Supplementary Figures and Tables from A TLR3 Ligand Reestablishes Chemotherapeutic Responses in the Context of FPR1 Deficiency

Author

Guido Kroemer, Erika Vacchelli, Jonathan G. Pol, Laurence Zitvogel, Jessica Zucman-Rossi, Pierre Laurent-Puig, Zoltan Szallasi, Suzette Delaloge, Fabrice André, Peter Vandenabeele, Federico Pietrocola, Maria Chiara Maiuri, Oliver Kepp, Cornelia Richter, Khady Mangane, Juliette Paillet, Gautier Stoll, Isabelle Martins, Francesca Castoldi, Giulia Cerrato, Allan Sauvat, Florent Ginhoux, Charles-Antoine Dutertre, Aymeric Silvin, Claire Mulot, Julien Taieb, Zsofia Sztupinszki, Sandy Adjemian, Liwei Zhao, Peng Liu, and Julie Le Naour

Abstract

Supplementary Figures and Tables

Published
2023
Full Text
View/download PDF

25. Table S4 from A TLR3 Ligand Reestablishes Chemotherapeutic Responses in the Context of FPR1 Deficiency

Author

Guido Kroemer, Erika Vacchelli, Jonathan G. Pol, Laurence Zitvogel, Jessica Zucman-Rossi, Pierre Laurent-Puig, Zoltan Szallasi, Suzette Delaloge, Fabrice André, Peter Vandenabeele, Federico Pietrocola, Maria Chiara Maiuri, Oliver Kepp, Cornelia Richter, Khady Mangane, Juliette Paillet, Gautier Stoll, Isabelle Martins, Francesca Castoldi, Giulia Cerrato, Allan Sauvat, Florent Ginhoux, Charles-Antoine Dutertre, Aymeric Silvin, Claire Mulot, Julien Taieb, Zsofia Sztupinszki, Sandy Adjemian, Liwei Zhao, Peng Liu, and Julie Le Naour

Abstract

Table S4

Published
2023
Full Text
View/download PDF

26. Data from A TLR3 Ligand Reestablishes Chemotherapeutic Responses in the Context of FPR1 Deficiency

Author

Guido Kroemer, Erika Vacchelli, Jonathan G. Pol, Laurence Zitvogel, Jessica Zucman-Rossi, Pierre Laurent-Puig, Zoltan Szallasi, Suzette Delaloge, Fabrice André, Peter Vandenabeele, Federico Pietrocola, Maria Chiara Maiuri, Oliver Kepp, Cornelia Richter, Khady Mangane, Juliette Paillet, Gautier Stoll, Isabelle Martins, Francesca Castoldi, Giulia Cerrato, Allan Sauvat, Florent Ginhoux, Charles-Antoine Dutertre, Aymeric Silvin, Claire Mulot, Julien Taieb, Zsofia Sztupinszki, Sandy Adjemian, Liwei Zhao, Peng Liu, and Julie Le Naour

Abstract

For anthracycline-based chemotherapy to be immunogenic, dying cancer cells must release annexin A1 (ANXA1) that subsequently interacts with the pattern recognition receptor, formyl peptide receptor 1 (FPR1), on the surface of dendritic cells (DC). Approximately 30% of individuals bear loss-of-function alleles of FPR1, calling for strategies to ameliorate their anticancer immune response. Here, we show that immunotherapy with a ligand of Toll-like receptor-3, polyinosinic:polycytidylic acid (pIC), restores the deficient response to chemotherapy of tumors lacking ANXA1 developing in immunocompetent mice or those of normal cancers growing in FPR1-deficient mice. This effect was accompanied by improved DC- and T-lymphocyte–mediated anticancer immunity. Of note, carcinogen-induced breast cancers precociously developed in FPR1-deficient mice as compared with wild-type controls. A similar tendency for earlier cancer development was found in patients carrying the loss-of-function allele of FPR1. These findings have potential implications for the clinical management of FPR1-deficient patients.Significance:The loss-of-function variant rs867228 in FPR1, harbored by approximately 30% of the world population, is associated with the precocious manifestation of breast, colorectal, esophageal, and head and neck carcinomas. pIC restores deficient chemotherapeutic responses in mice lacking Fpr1, suggesting a personalized strategy for compensating for the FPR1 defect.This article is highlighted in the In This Issue feature, p. 211

Published
2023
Full Text
View/download PDF

27. Nanoparticles releasing immunogenic cell death inducers upon near-infrared light exposure

Author

Oliver Kepp, Giulia Cerrato, Allan Sauvat, and Guido Kroemer

Subjects
Infrared Rays, Immunology, Antineoplastic Agents, Immunogenic Cell Death, Mice, Editorial, Oncology, Neoplasms, Tumor Microenvironment, Nanoparticles, Immunology and Allergy, Animals, Indoleamine-Pyrrole 2,3,-Dioxygenase, Immunotherapy, Transcytosis, Copper
Abstract

Cancer photo-immunotherapy (CPIT) as an ideal strategy can rapidly release hostile signals by appropriate dosage of focal laser irradiation to unmask primary tumor immunogenicity and can activate adaptive immunity to control distant metastases. However, many factors, including disordered immunometabolism, poor penetration of photothermal agents and immuno-regulators, inadequate laser penetration into the deep tumor region, restrict the therapeutic outcomes of CPIT. Here, a second near-infrared window (NIR-II) photo-immunometabolic cancer therapy (PICT) by a programmed raspberry-structured nanoadjuvant (PRN

Published
2022

28. ACBP/DBI protein neutralization confers autophagy-dependent organ protection through inhibition of cell loss, inflammation, and fibrosis

Author

Omar Motiño, Flavia Lambertucci, Gerasimos Anagnostopoulos, Sijing Li, Jihoon Nah, Francesca Castoldi, Laura Senovilla, Léa Montégut, Hui Chen, Sylvère Durand, Mélanie Bourgin, Fanny Aprahamian, Nitharsshini Nirmalathasan, Karla Alvarez-Valadez, Allan Sauvat, Vincent Carbonnier, Mojgan Djavaheri-Mergny, Federico Pietrocola, Junichi Sadoshima, Maria Chiara Maiuri, Isabelle Martins, Guido Kroemer, Agence Nationale de la Recherche (France), Institut Universitaire de France, Fondation Leducq, Université de Paris, CSIC-UVA - Instituto de Biología y Genética Molecular (IBGM), and Junta de Castilla y León

Subjects
Diazepam Binding Inhibitor, Inflammation, Multidisciplinary, Diazepam, Indoleacetic Acids, Fatty Acids, Antibodies, Monoclonal, Receptors, GABA-A, Acyl-CoA binding protein, Fibrosis, Antioxidants, Choline, Mice, Methionine, Concanavalin A, Autophagy, Myocardium infarction, Animals, Coenzyme A, Carrier Proteins, Carbon Tetrachloride, Non-alcoholic steatohepatitis, Acetaminophen, Autoantibodies
Abstract

Acyl-coenzyme A (CoA)–binding protein (ACBP), also known as diazepam-binding inhibitor (DBI), is an extracellular feedback regulator of autophagy. Here, we report that injection of a monoclonal antibody neutralizing ACBP/DBI (α-DBI) protects the murine liver against ischemia/reperfusion damage, intoxication by acetaminophen and concanavalin A, and nonalcoholic steatohepatitis caused by methionine/choline-deficient diet as well as against liver fibrosis induced by bile duct ligation or carbon tetrachloride. α-DBI downregulated proinflammatory and profibrotic genes and upregulated antioxidant defenses and fatty acid oxidation in the liver. The hepatoprotective effects of α-DBI were mimicked by the induction of ACBP/DBI-specific autoantibodies, an inducible Acbp/Dbi knockout or a constitutive Gabrg2F77I mutation that abolishes ACBP/DBI binding to the GABAA receptor. Liver-protective α-DBI effects were lost when autophagy was pharmacologically blocked or genetically inhibited by knockout of Atg4b. Of note, α-DBI also reduced myocardium infarction and lung fibrosis, supporting the contention that it mediates broad organ-protective effects against multiple insults., We thank the core facilities of Centre de Recherche des Cordeliers and Gustave Roussy for technical support. G.K. is supported by the Ligue contre le Cancer (equipe labellisee); Agence National de la Recherche (ANR) – Projets blancs; AMMICa US23/CNRS UMS3655; Association pour la Recherche sur le Cancer; Association “Ruban Rose”; Cancerop^ole Ile-de-France; Fondation pour la Recherche Medicale (FRM); a donation by Elior; Equipex Onco-Pheno-Screen; European Joint Programme on Rare Diseases; Gustave Roussy Odyssea, the European Union Horizon 2020 Projects Oncobiome and Crimson (No. 101016923); Fondation Carrefour; Institut National du Cancer; Inserm (Heterogeneite des tumeurs dans leur microenvironnement); Institut Universitaire de France; LabEx Immuno-Oncology (ANR-18- IDEX-0001); the Leducq Foundation; a Cancer Research Accelerating Scientific Platforms and Innovative Research Award from the Mark Foundation;, the Recherche Hospitalo-Universitaire Torino Lumiere; Seerave Foundation; SIRIC Stratified Oncology Cell DNA Repair and Tumor Immune Elimination; and SIRIC Cancer Research and Personalized Medicine. This study contributes to the IdEx Universite de Paris ANR-18-IDEX-0001. G.A. is supported by the FRM. L.S. is supported by Beatriz Galindo senior program of the Spanish Ministry of Universities; Strategic Program “Instituto de Biologıa y Genetica Molecular (IBGM), Junta de Castilla y Leon” (Ref. CCVC8485); and Internationalisation Project of the “Unidad de Excelencia IBGM of Valladolid” (Ref. CL-EI-2021).

Published
2022

29. Assessment of transcription inhibition as a characteristic of immunogenic cell death

Author

Giulia, Cerrato, Juliette, Humeau, Allan, Sauvat, Oliver, Kepp, and Guido, Kroemer

Subjects
RNA, Click Chemistry, Immunogenic Cell Death
Abstract

Anticancer drugs that suppress DNA-to-RNA transcription are particularly efficient in stimulating immunogenic cell death and hence eradicate malignant cells in a way that they will ignite an antitumor immune response. This is therapeutically relevant as it allows treatment response to last beyond drug discontinuation. For this reason, it is important to measure transcription inhibition in a precise fashion. Here, we detail two complementary assays for the assessment of transcription inhibition, one that detects the physical separation of fibrillarin and nucleolin by two-color immunofluorescence and another that measures the diminution of incorporated 5-ethynyl uridine (EU) into RNA, as revealed by click chemistry and the per-cell-intensity of a fluorescent signal.

Published
2022

30. Assessment of type I interferon responses as a feature of immunogenic cell death

Author

Sabrina, Forveille, Allan, Sauvat, Shuai, Zhang, Liwei, Zhao, Guido, Kroemer, and Oliver, Kepp

Subjects
Drug Discovery, Interferon Type I, Immunogenic Cell Death, High-Throughput Screening Assays
Abstract

The radiochemotherapy- or chemotherapy-induced stimulation of immunogenic cell death (ICD) affecting malignant cells ignites antitumor immune responses that are clinically relevant as they allow to achieve durable responses beyond treatment discontinuation. The mechanistic exploration of ICD and the discovery of agents and interventions that are endowed with the capacity to elicit ICD is of the utmost importance. Here, we describe an assay for the assessment of type I interferon (IFN) production, which is one of the salient features of ICD. Biosensor cells that express GFP under the control of the IFN-inducible MX dynamin like GTPase 1 (MX1) gene promoter are employed, and the fluorescent signal is assessed by automated microscopy. The described workflow is automation-friendly, rendering it compatible with high-throughput screening (HTS) for drug discovery.

Published
2022

31. Cancer cell-autonomous overactivation of PARP1 compromises immunosurveillance in non-small cell lung cancer

Author

Pan Juncheng, Adrien Joseph, Antoine Lafarge, Isabelle Martins, Florine Obrist, Jonathan Pol, Ester Saavedra, Sijing Li, Allan Sauvat, Giulia Cerrato, Sarah Lévesque, Marion Leduc, Oliver Kepp, Sylvère Durand, Fanny Aprahamian, Nitharsshini Nirmalathansan, Judith Michels, Guido Kroemer, and Maria Castedo

Subjects
Pharmacology, Cancer Research, Lung Neoplasms, Immunology, Poly(ADP-ribose) Polymerase Inhibitors, Mice, Oncology, Monitoring, Immunologic, Carcinoma, Non-Small-Cell Lung, Tumor Microenvironment, Molecular Medicine, Immunology and Allergy, Animals, Cisplatin
Abstract

BackgroundHigh activity of poly(ADP-ribose) polymerase-1 (PARP1) in non-small cell lung cancer (NSCLC) cells leads to an increase in immunohistochemically detectable PAR, correlating with poor prognosis in patients with NSCLC, as well as reduced tumor infiltration by cytotoxic T lymphocytes (CTLs). Intrigued by this observation, we decided to determine whether PARP1 activity in NSCLC cells may cause an alteration of anticancer immunosurveillance.MethodsContinuous culture of mouse NSCLC cells in the presence of cisplatin led to the generation of cisplatin-resistant PARhigh clones. As compared with their parental controls, such PARhigh cells formed tumors that were less infiltrated by CTLs when they were injected into immunocompetent mice, suggesting a causative link between high PARP1 activity and compromised immunosurveillance. To confirm this cause-and-effect relationship, we used CRISPR/Cas9 technology to knock out PARP1 in two PARhigh NSCLC mouse cell lines (Lewis lung cancer [LLC] and tissue culture number one [TC1]), showing that the removal of PARP1 indeed restored cisplatin-induced cell death responses.ResultsPARP1 knockout (PARP1KO) cells became largely resistant to the PARP inhibitor niraparib, meaning that they exhibited less cell death induction, reduced DNA damage response, attenuated metabolic shifts and no induction of PD-L1 and MHC class-I molecules that may affect their immunogenicity. PARhigh tumors implanted in mice responded to niraparib irrespective of the presence or absence of T lymphocytes, suggesting that cancer cell-autonomous effects of niraparib dominate over its possible immunomodulatory action. While PARhigh NSCLC mouse cell lines proliferated similarly in immunocompetent and T cell-deficient mice, PARP1KO cells were strongly affected by the presence of T cells. PARP1KO LLC tumors grew more quickly in immunodeficient than in immunocompetent mice, and PARP1KO TC1 cells could only form tumors in T cell-deficient mice, not in immunocompetent controls. Importantly, as compared with PARhigh controls, the PARP1KO LLC tumors exhibited signs of T cell activation in the immune infiltrate such as higher inducible costimulator (ICOS) expression and lower PD-1 expression on CTLs.ConclusionsThese results prove at the genetic level that PARP1 activity within malignant cells modulates the tumor microenvironment.

Published
2022

32. Oleate-induced aggregation of LC3 at the trans-Golgi network is linked to a protein trafficking blockade

Author

Guido Kroemer, Oliver Kepp, Shuai Zhang, Peng Liu, Giulia Cerrato, Wei Xie, Allan Sauvat, Liwei Zhao, Kevin Müller, Marion Leduc, and Juliette Humeau

Subjects
0301 basic medicine, Lactosylceramides, Endogeny, Mice, 03 medical and health sciences, symbols.namesake, 0302 clinical medicine, In vivo, Autophagy, Animals, Humans, Secretion, Molecular Biology, biology, Chemistry, Endoplasmic reticulum, Cell Biology, Golgi apparatus, biology.organism_classification, Autophagic Punctum, In vitro, Cell biology, Protein Transport, 030104 developmental biology, Secretory protein, Vesicular stomatitis virus, 030220 oncology & carcinogenesis, symbols, Oleic Acid, trans-Golgi Network
Abstract

Oleate, the most abundantly occurring cis-unsaturated fatty acid, has the particularity to induce the accumulation of MAP1LC3B/LC3 (microtubule associated protein 1 light chain 3 beta) at the trans-Golgi apparatus. A genome-wide RNA interference screen designed to identify the mechanisms of this LC3 redistribution led to the identification of a BECN1-PIK3C3-independent pathway that, however, requires the ATG12–ATG5 and ATG7-dependent conjugation system, and several genes/proteins involved in endoplasmic reticulum (ER)-to-Golgi anterograde protein transport, as well as the unfolded protein response, including the integrated stress response that results in the phosphorylation of EIF2A/eIF2α (eukaryotic translation initiation factor 2A). Functional experiments revealed that oleate blocks conventional protein secretion, stalling the process at the level of the trans-Golgi network. Oleate-induced blockade of protein secretion occurred even after depletion of ATG5, suggesting that it does not rely on the recruitment of LC3 to the Golgi apparatus (which does require ATG5). Rather, it appears that oleate and other pharmacological inhibitors of protein secretion with a similar mode of action provoke a perturbation of the trans-Golgi compartment that secondarily results in the local enrichment of LC3.

Published
2020
Full Text
View/download PDF

33. Multi-scale generative adversarial network for improved evaluation of cell–cell interactions observed in organ-on-chip experiments

Author

C. Di Natale, Giulia Cerrato, Davide Di Giuseppe, Arianna Mencattini, Fabrizio Mattei, Allan Sauvat, Joanna Filippi, Guido Kroemer, Giovanna Schiavoni, A. De Ninno, Erika Vacchelli, Eugenio Martinelli, Luca Businaro, Paola Casti, Michele D'Orazio, and Maria Colomba Comes

Subjects
Video prediction, 0209 industrial biotechnology, Computer science, Distributed computing, Cell, ComputingMethodologies_IMAGEPROCESSINGANDCOMPUTERVISION, Predictive capability, Context (language use), 02 engineering and technology, Settore ING-INF/07, Organ-on-a-chip, 020901 industrial engineering & automation, Artificial Intelligence, Microscopy, 0202 electrical engineering, electronic engineering, information engineering, medicine, Scale (chemistry), Biomedical application, Generative adversarial network (GAN), Time-lapse microscopy, Volume (computing), Deep learning, Frame rate, Photobleaching, Visualization, medicine.anatomical_structure, 020201 artificial intelligence & image processing, Generative adversarial network, Software
Abstract

Organs On a Chip (OOCs) represent a sophisticated approach for exploring biological mechanisms and developing therapeutic agents. In conjunction with high-quality time-lapse microscopy (TLM), OOCs allow for the visualization of reconstituted complex biological processes, such as multi-cell-type migration and cell–cell interactions. In this context, increasing the frame rate is desirable to reconstruct accurately cell-interaction dynamics. However, a trade-off between high resolution and carried information content is required to reduce the overall data volume. Moreover, high frame rates increase photobleaching and phototoxicity. As a possible solution for these problems, we report a new hybrid-imaging paradigm based on the integration of OOC/TLMs with a Multi-scale Generative Adversarial Network (GAN) predicting interleaved video frames with the aim to provide high-throughput videos. We tested the performance of the predictive capability of GAN on synthetic videos, as well as on real OOC experiments dealing with tumor–immune cell interactions. The proposed approach offers the possibility to acquire a reduced number of high-quality TLM images without any major loss of information on the phenomena under investigation.

Published
2020
Full Text
View/download PDF

34. Escherichia coli-Specific CXCL13-Producing TFH Are Associated with Clinical Efficacy of Neoadjuvant PD-1 Blockade against Muscle-Invasive Bladder Cancer

Author

Anne-Gaëlle Goubet, Leonardo Lordello, Carolina Alves Costa Silva, Isabelle Peguillet, Marianne Gazzano, Maxime Descartes Mbogning-Fonkou, Cassandra Thelemaque, Cédric Lebacle, Constance Thibault, François Audenet, Géraldine Pignot, Gwenaelle Gravis, Carole Helissey, Luca Campedel, Morgan Roupret, Evanguelos Xylinas, Idir Ouzaid, Agathe Dubuisson, Marine Mazzenga, Caroline Flament, Pierre Ly, Virginie Marty, Nicolas Signolle, Allan Sauvat, Thomas Sbarrato, Mounia Filahi, Caroline Davin, Gabriel Haddad, Jacques Bou Khalil, Camille Bleriot, François-Xavier Danlos, Garett Dunsmore, Kevin Mulder, Aymeric Silvin, Thibault Raoult, Baptiste Archambaud, Shaima Belhechmi, Ivo Gomperts Boneca, Nadège Cayet, Maryse Moya-Nilges, Adeline Mallet, Romain Daillere, Etienne Rouleau, Camelia Radulescu, Yves Allory, Jacques Fieschi, Mathieu Rouanne, Florent Ginhoux, Gwénaël Le Teuff, Lisa Derosa, Aurélien Marabelle, Jeroen Van Dorp, Nick Van Dijk, Michiel S. Van Der Heijden, Benjamin Besse, Fabrice Andre, Miriam Merad, Guido Kroemer, Jean-Yves Scoazec, Laurence Zitvogel, Yohann Loriot, Université Paris-Saclay, Institut Gustave Roussy (IGR), Immunologie anti-tumorale et immunothérapie des cancers (ITIC), Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay, NF-kappaB, Différenciation et Cancer (OncokappaB (URP_7324)), Université Paris Cité (UPCité), Immunologie intégrative des tumeurs et immunothérapie des cancers (INTIM), Institut Curie [Paris], Centre d'Immunologie et des Maladies Infectieuses (CIMI), Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Petites Molécules de neuroprotection, neurorégénération et remyélinisation, Université Paris-Sud - Paris 11 (UP11)-Institut National de la Santé et de la Recherche Médicale (INSERM), Hôpital Européen Georges Pompidou [APHP] (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO), Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC), Centre de Recherche en Cancérologie de Marseille (CRCM), Aix Marseille Université (AMU)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Hôpital d'Instruction des Armées Begin, Service de Santé des Armées, CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), AP-HP - Hôpital Bichat - Claude Bernard [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Immunologie des tumeurs et immunothérapie (UMR 1015), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPCité), Microbes évolution phylogénie et infections (MEPHI), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Institut Hospitalier Universitaire Méditerranée Infection (IHU Marseille), Département d’Innovation Thérapeutique et essais précoces [Gustave Roussy] (DITEP), Direction de la recherche clinique [Gustave Roussy], Service de biostatistique et d'épidémiologie (SBE), Institut Gustave Roussy (IGR)-Institut Gustave Roussy (IGR), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Département de médecine oncologique [Gustave Roussy], Département de biologie et pathologie médicales [Gustave Roussy], Oncologie gynécologique, ANR-16-RHUS-0008, ANR-21-RHUS-0017, Bristol-Myers Squibb, BMS, Pfizer, Astellas Pharma US, APUS, AstraZeneca, Genentech, Merck, Roche, Gilead Sciences, Meso Scale Diagnostics, MSD, Janssen Pharmaceuticals, Merck Sharp and Dohme, MSD, Horizon 2020 Framework Programme, H2020: 19-CE15-0029-01, 825410, Clovis Oncology, Seerave Foundation, Fondation Philanthropia, Advanced Accelerator Applications, AAA, Agence Nationale de la Recherche, ANR: ANR-10-LABX-62-IBEID, Fondation pour la Recherche Médicale, FRM, Daiichi-Sankyo, Fondation ARC pour la Recherche sur le Cancer, ARC, Ligue Contre le Cancer, Institut Universitaire de France, IUF, Institut National Du Cancer, INCa, Cancéropôle Ile de France, Association Française d'Urologie, AFU, Labex Immuno-Oncology, The trial was conducted by the French Genitourinary Group (GETUG) and funded by MSD, which provided the drug. This study was approved by the ethics committee CPP Est-III in December 2017 and the French National Agency for the Safety of Medicines and Health Products (ANSM) in November 2017, and was conducted in accordance with the protocols and Good Clinical Practice Guidelines defined by the International Conference for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use and the principles of the Declaration of Helsinki., C. Alves Costa Silva reports grants from MSD Avenir Foundation during the conduct of the study. C. Thibault reports personal fees and nonfinancial support from Pfizer, Merck, MSD, Janssen, and Ipsen, grants, personal fees, and nonfinancial support from AstraZeneca, We thank pathologists, nurses, and clinical research associates from Hôpital Européenn George Pompidou, Hôpital Begin, Institut Paoli-Calmettes, Hôpital Bichat, and Hôpital Pitié-Salpétrière for their participation in the PANDORE clinical trial. We are thankful to the flow and mass cytometry facility team of Gustave Roussy (Philippe Rameau and Cyril Catelain). We thank Fluidigm for their support. We are grateful for support for equipment from the French Government Programme Investissements d’Avenir France BioImag-ing (FBI, No. ANR-10-INSB-04-01) and the French Government (Agence Nationale de la Recherche) Investissement d’Avenir program, Laboratoire d’Excellence 'Integrative Biology of Emerging Infectious Diseases' (ANR-10-LABX-62-IBEID). A.-G. Goubet was supported by Fondation pour la Recherche Médicale. C. Alves Costa Silva was supported by MSD Avenir. F.-X. Danlos was supported by Fondation Philantropia. M. Roupret was supported by Fondation Foch and the Association Française d’Urologie (AFU). L. Zitvogel was funded by the RHU Torino Lumière (ANR-16-RHUS-0008). L. Zitvogel and L. Derosa were supported by RHU5 'ANR-21-RHUS-0017' IMMUNOLIFE, SIRIC Stratified Oncology Cell DNA Repair and Tumor Immune Elimination (SOCRATE), as well as the SIGN’IT ARC foundation. L. Zitvogel was supported by European Union’s Horizon 2020 research and innovation programme under grant agreement number 825410 [project acronym: ONCOBIOME, project title: Gut OncoMicrobiome Signatures (GOMS) associated with cancer incidence, prognosis, and prediction of treatment response]. L. Zitvogel also received an ANR grant–French-German Ileobiome 19-CE15-0029-01. L. Zitvogel and G. Kroemer received a donation from the Seerave Foundation. L. Zitvogel and G. Kroemer were supported by the Ligue contre le Cancer (équipe labelisée), ANR projets blancs, Cancéropôle Ile-de-France, Fondation pour la Recherche Médicale (FRM), a donation by Elior, Institut National du Cancer (INCa), Inserm (HTE), Institut Universitaire de France, the LabEx Immuno-Oncology, and FHU CARE, Dassault, and Badinter Philantropia. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact., Bristol Myers Squibb, and Sanofi, and personal fees from Astellas and AAA during the conduct of the study, as well as personal fees and nonfinancial support from Pfizer, Merck, MSD, Janssen, and Ipsen, grants, personal fees, and nonfinancial support from AstraZeneca, Bristol Myers Squibb, and Sanofi, and personal fees from Astellas and AAA outside the submitted work. F. Audenet reports personal fees from Astellas, Urodiag, Vitadx, and Bristol Myers Squibb, and nonfinancial support from Ipsen outside the submitted work. G. Gravis reports grants from Bristol Myers Squibb, and other support from MSD, Bristol Myers Squibb, and Merck–Pfizer alliance outside the submitted work. C. Helissey reports personal fees from Janssen-Cilag, Roche, Bayer, AstraZeneca, and Astellas outside the submitted work. L. Cam-pedel reports personal fees from MSD, Pfizer, Bristol Myers Squibb, and Bayer outside the submitted work. T. Sbarrato reports personal fees from Veracyte during the conduct of the study. T. Raoult reports grants, personal fees, and nonfinancial support from Merck Sharp & Dohme during the conduct of the study. E. Rouleau reports grants from AstraZeneca, Roche, Clovis, Bristol Myers Squibb, and MSD outside the submitted work. Y. Allory reports other support from Astra-Zeneca, MSD, and Bristol Myers Squibb outside the submitted work. J. Fieschi reports personal fees from Veracyte during the conduct of the study. A. Marabelle reports grants, personal fees, nonfinancial support, and other support from MSD, Bristol Myers Squibb, and AstraZeneca, and personal fees, nonfinancial support, and other support from Roche/Genentech and Pfizer outside the submitted work. J. Van Dorp reports other support from Bristol Myers Squibb during the conduct of the study. M.S. van der Heijden reports grants from Bristol Myers Squibb during the conduct of the study, as well as grants and personal fees from Bristol Myers Squibb, Roche, and AstraZeneca, grants from 4SC, and personal fees from MSD/Merck, Janssen, Pfizer, and Seagen outside the submitted work. B. Besse reports grants from 4D Pharma, AbbVie, Amgen, Aptitude Health, AstraZeneca, BeiGene, Blueprint Medicines, Boehringer Ingelheim, Celgene, Cergentis, Chu-gai Pharmaceutical, Cristal Therapeutics, Daiichi Sankyo, Eli Lilly, Eisai, Genzyme Corporation, GSK, Inivata, Ipsen, Janssen, Onxeo, OSE Immunotherapeutics, Pfizer, Roche/Genentech, Sanofi, Takeda, Tolero Pharmaceuticals, and Turning Point Therapeutics during the conduct of the study. F. Andre reports grants and other support from AstraZeneca and Daiichi Sankyo, grants from Lilly and Sanofi, and other support from Novartis and Relay outside the submitted work. M. Merad reports grants and personal fees from Regeneron, personal fees from Compugen, Morphic Therapeutics, Myeloid Therapeutics, Nirogy, DrenBio, Oncoresponse, Asherbio, Pionyr, Owkin, and Larkspur, other support from Innate Pharma, Genenta, DBV, and OSE Immunotherapeutics, and grants from Boehringer outside the submitted work. G. Kroemer reports grants from Daiichi Sankyo, Eleor, Kaleido, Lytix Pharma, PharmaMar, Osasuna Therapeutics, Samsara Therapeutics, Sanofi, Sotio, Tollys, Vascage, and Vasculox/Tioma, and personal fees from Reithera outside the submitted work, is on the Board of Directors for Bristol Myers Squibb Foundation France, and is a scientific cofounder of EverImmune, Osasuna Therapeutics, Samsara Therapeutics, and Therafast Bio. L. Zitvogel reports grants and personal fees from EverImmune, and grants from 9 Meters and Pileje during the conduct of the study, grants from Daiichi Sankyo outside the submitted work, and a patent for B220028EPA pending. Y. Loriot reports grants from MSD and personal fees from MSD during the conduct of the study, personal fees from Bristol Myers Squibb, Pfizer, Merck Serono, AstraZeneca, Seattle Genetics, Gilead, Taiho, and Astel-las, grants and personal fees from Janssen, and grants from Roche and Celsius outside the submitted work, and a patent for EP2305181.4 pending. No disclosures were reported by the other authors., and FHU CARE, Dassault, and Badinter Philantropia., ANR-16-RHUS-0008,LUMIERE,LUMIERE(2016), and ANR-21-RHUS-0017,IMMUNOLIFE,Microbiota-centered interventions to solve antibiotics-induced primary resistance to immune checkpoint inhibitors(2021)

Subjects
[SDV]Life Sciences [q-bio], Muscles, Programmed Cell Death 1 Receptor, T-Lymphocytes, Helper-Inducer, Chemokine CXCL13, B7-H1 Antigen, Neoadjuvant Therapy, Treatment Outcome, Oncology, Urinary Bladder Neoplasms, Immunoglobulin G, Escherichia coli, Humans, Immune Checkpoint Inhibitors
Abstract

Biomarkers guiding the neoadjuvant use of immune-checkpoint blockers (ICB) are needed for patients with localized muscle-invasive bladder cancers (MIBC). Profiling tumor and blood samples, we found that follicular helper CD4+ T cells (TFH) are among the best therapeutic targets of pembrolizumab correlating with progression-free survival. TFH were associated with tumoral CD8 and PD-L1 expression at baseline and the induction of tertiary lymphoid structures after pembrolizumab. Blood central memory TFH accumulated in tumors where they produce CXCL13, a chemokine found in the plasma of responders only. IgG4+CD38+ TFH residing in bladder tissues correlated with clinical benefit. Finally, TFH and IgG directed against urothelium-invasive Escherichia coli dictated clinical responses to pembrolizumab in three independent cohorts. The links between tumor infection and success of ICB immunomodulation should be prospectively assessed at a larger scale. Significance: In patients with bladder cancer treated with neoadjuvant pembrolizumab, E. coli–specific CXCL13 producing TFH and IgG constitute biomarkers that predict clinical benefit. Beyond its role as a biomarker, such immune responses against E. coli might be harnessed for future therapeutic strategies. This article is highlighted in the In This Issue feature, p. 2221

Published
2022
Full Text
View/download PDF

37. Control of lysosomal-mediated cell death by the pH-dependent calcium channel RECS1

Author

Claudio Hetz, Alvaro Glavic, Philippe Pihán, Amaury Pupo, Suvi Saarnio, Giovanni Quarato, Sandra Sofía Edwards-Jorquera, Diego A. Rodriguez, Daniela De Giorgis, Guido Kroemer, Oliver Kepp, Nicolas Demaurex, Karen Castillo, Eija Jokitalo, Janina Borgonovo, Sabrina Forveille, Amado Carreras-Sureda, Fernanda Lourido, Miguel L. Concha, Paula Nunes-Hasler, Fernanda Lisbona, Douglas R. Green, Anthony Letai, Hery Urra, Alfredo Sagredo, Allan Sauvat, Ramon Latorre, Helena Vihinen, Universidad de Chile = University of Chile [Santiago] (UCHILE), Université de Genève (UNIGE), Universidad de Valparaiso [Chile], Institut Universitaire de France (IUF), Ministère de l'Education nationale, de l’Enseignement supérieur et de la Recherche (M.E.N.E.S.R.), Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université de Paris (UP), Institut Gustave Roussy (IGR), University of Helsinki, St Jude Children's Research Hospital, Dana-Farber Cancer Institute [Boston], Brigham & Women’s Hospital [Boston] (BWH), Harvard Medical School [Boston] (HMS), Hôpital Européen Georges Pompidou [APHP] (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO), Chinese Academy of Sciences [Beijing] (CAS), Karolinska Institutet [Stockholm], Universidad Catolica Del Maule, Université de Genève = University of Geneva (UNIGE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPCité), Métabolisme, Cancer et Immunité (CRC - UMR_S 1138), Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPCité)-Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPCité)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Helsingin yliopisto = Helsingfors universitet = University of Helsinki, HAL-SU, Gestionnaire, Molecular and Integrative Biosciences Research Programme, Institute of Biotechnology, Electron Microscopy, and Biosciences

Subjects
Programmed cell death, STRESS, BI-1, [SDV]Life Sciences [q-bio], MEMBRANE PERMEABILIZATION, Ph dependent, ddc:616.07, ANTI-APOPTOTIC PROTEINS, Inhibitory postsynaptic potential, CA2+, 03 medical and health sciences, 0302 clinical medicine, FLUORESCENCE, ddc:612, 030304 developmental biology, 0303 health sciences, Multidisciplinary, UNFOLDED PROTEIN RESPONSE, Chemistry, Calcium channel, SciAdv r-articles, Life Sciences, Cell Biology, NEGATIVE REGULATOR, Cell biology, [SDV] Life Sciences [q-bio], BAX INHIBITOR-1, ER, 030220 oncology & carcinogenesis, 1182 Biochemistry, cell and molecular biology, Biomedicine and Life Sciences, Research Article
Abstract

Description, RECS1, a member of the TMBIM family, triggers lysosomal membrane permeabilization in cells undergoing lysosomal stress., Programmed cell death is regulated by the balance between activating and inhibitory signals. Here, we have identified RECS1 (responsive to centrifugal force and shear stress 1) [also known as TMBIM1 (transmembrane BAX inhibitor motif containing 1)] as a proapoptotic member of the TMBIM family. In contrast to other proteins of the TMBIM family, RECS1 expression induces cell death through the canonical mitochondrial apoptosis pathway. Unbiased screening indicated that RECS1 sensitizes cells to lysosomal perturbations. RECS1 localizes to lysosomes, where it regulates their acidification and calcium content, triggering lysosomal membrane permeabilization. Structural modeling and electrophysiological studies indicated that RECS1 is a pH-regulated calcium channel, an activity that is essential to trigger cell death. RECS1 also sensitizes whole animals to stress in vivo in Drosophila melanogaster and zebrafish models. Our results unveil an unanticipated function for RECS1 as a proapoptotic component of the TMBIM family that ignites cell death programs at lysosomes.

Published
2021
Full Text
View/download PDF

38. Autoimmunity affecting the biliary tract fuels the immunosurveillance of cholangiocarcinoma

Author

Franck Letourneur, Jérémy Augustin, Mark J. Kurth, Julie Le Naour, Sylvie Lachkar, Pamela Caudana, Eliane Piaggio, Youra Kim, Sarah Levesque, Jie S. Zhu, Guido Kroemer, Jonathan Pol, Bouchra Lekbaby, Andrea Checcoli, Patrick S.C. Leung, Allan Sauvat, Shashi Gujar, Agathe Delaune, Chantal Housset, Pierre de la Grange, Paule Opolon, Laura Fouassier, Noémie Robil, Laurence Zitvogel, Patrick Soussan, Juliette Paillet, Céleste Plantureux, Jimena Tosello Boari, M. Eric Gershwin, Isabelle Martins, Maria Chiara Maiuri, Cédric Coulouarn, Gautier Stoll, Norma Bloy, Gwennhael Autret, Juliette Hamroune, Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université de Paris (UP), Institut Gustave Roussy (IGR), Institut Curie [Paris], Immunité et cancer (U932), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Curie [Paris], Cancer et génome: Bioinformatique, biostatistiques et épidémiologie d'un système complexe, Institut Curie [Paris]-MINES ParisTech - École nationale supérieure des mines de Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM), GenoSplice [Paris], Institut Cochin (IC UM3 (UMR 8104 / U1016)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Paris-Centre de Recherche Cardiovasculaire (PARCC (UMR_S 970/ U970)), Hôpital Européen Georges Pompidou [APHP] (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP), University of California [Los Angeles] (UCLA), University of California, Centre de Recherche Saint-Antoine (CR Saint-Antoine), Sorbonne Université (SU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Saint-Antoine [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Dalhousie University [Halifax], Chemistry, Oncogenesis, Stress and Signaling (COSS), Institut National de la Santé et de la Recherche Médicale (INSERM)-CRLCC Eugène Marquis (CRLCC)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES), CRLCC Eugène Marquis (CRLCC), Université Paris-Saclay, Immunologie des tumeurs et immunothérapie (UMR 1015), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Gustave Roussy (IGR)-Université Paris-Sud - Paris 11 (UP11), Centre d'Investigation Clinique en Biotherapie des cancers (CIC 1428 , CBT 507 ), Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Karolinska University Hospital [Stockholm], Association pour la lutte contre les maladies inflammatoires du foie et des voies biliaires, Dalhousie Medical Research Foundation, Agence National de la Recherche, ERA-Net for Research on Rare Diseases, Association pour la recherche sur le cancer, Cancéropôle Ile-de-France, Chancellerie des universités de Paris, Fondation pour la Recherche Médicale, Elior, European Research Area Network on Cardiovascular Diseases, Gustave Roussy Odyssea, European Union Horizon 2020, Canadian Cancer Society, Project Oncobiome, Fondation Carrefour, GDW20171100085, High-end Foreign Expert Program in China, Institut National du Cancer, Inserm, Institut Universitaire de France, LeDucq Foundation, LabEx, Recherche Hospitalo-Universitaire Torino Lumière, Seerave Foundation, Canadian Institutes of Health Research, Site de Recherche intégrée sur le Cancer Stratified Oncology Cell DNA Repair and Tumor Immune Elimination, Cancer Research and Personalized Medicine, Multi-Organism Institute (ITMO) Aviesan Cancer, Association Française d'Hépatologie, Site de Recherche intégrée sur le Cancer Cancer Research and Personalized Medicine, Ligue contre le Cancer, École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPCité), Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Mines Paris - PSL (École nationale supérieure des mines de Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), University of California (UC), Centre de Recherche Saint-Antoine (CRSA), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Université de Rennes (UR)-CRLCC Eugène Marquis (CRLCC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), and Institut Gustave Roussy (IGR)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
CD4-Positive T-Lymphocytes, Cholangitis, Autoimmunity, CD8-Positive T-Lymphocytes, Inbred C57BL, medicine.disease_cause, Medical and Health Sciences, Malignant transformation, Cholangiocarcinoma, Mice, 0302 clinical medicine, Immunologic, Neoplasms, 2.1 Biological and endogenous factors, Immunology and Allergy, Medicine, Aetiology, Cancer, 0303 health sciences, Tumor, Liver Disease, Forkhead Transcription Factors, 3. Good health, Immunosurveillance, medicine.anatomical_structure, Liver, Biliary tract, 030220 oncology & carcinogenesis, Hepatocellular carcinoma, cardiovascular system, Cytokines, Female, Biotechnology, Liver Cancer, Monitoring, T cell, Chronic Liver Disease and Cirrhosis, Immunology, Context (language use), [SDV.CAN]Life Sciences [q-bio]/Cancer, Autoimmune Disease, digestive system, Cell Line, Primary sclerosing cholangitis, Experimental, 03 medical and health sciences, Rare Diseases, Monitoring, Immunologic, Cell Line, Tumor, parasitic diseases, Animals, cardiovascular diseases, Digestive Diseases - (Gallbladder), 030304 developmental biology, business.industry, Neoplasms, Experimental, medicine.disease, digestive system diseases, Mice, Inbred C57BL, Bile Duct Neoplasms, Cancer research, Digestive Diseases, business
Abstract

International audience; Cholangiocarcinoma (CCA) results from the malignant transformation of cholangiocytes. Primary sclerosing cholangitis (PSC) and primary biliary cholangitis (PBC) are chronic diseases in which cholangiocytes are primarily damaged. Although PSC is an inflammatory condition predisposing to CCA, CCA is almost never found in the autoimmune context of PBC. Here, we hypothesized that PBC might favor CCA immunosurveillance. In preclinical murine models of cholangitis challenged with syngeneic CCA, PBC (but not PSC) reduced the frequency of CCA development and delayed tumor growth kinetics. This PBC-related effect appeared specific to CCA as it was not observed against other cancers, including hepatocellular carcinoma. The protective effect of PBC was relying on type 1 and type 2 T cell responses and, to a lesser extent, on B cells. Single-cell TCR/RNA sequencing revealed the existence of TCR clonotypes shared between the liver and CCA tumor of a PBC host. Altogether, these results evidence a mechanistic overlapping between autoimmunity and cancer immunosurveillance in the biliary tract.

Published
2021
Full Text
View/download PDF

39. High throughput screening for autophagy

Author

Sabrina, Forveille, Marion, Leduc, Allan, Sauvat, Giulia, Cerrato, Guido, Kroemer, and Oliver, Kepp

Subjects
Drug Discovery, Green Fluorescent Proteins, Autophagy, Drug Evaluation, Preclinical, Microtubule-Associated Proteins, High-Throughput Screening Assays
Abstract

Robotized high throughput screening allows for the assessment of autophagy in a large number of samples. Here, we describe a drug discovery platform for the phenotypic identification of novel autophagy inducers by means of automated cell biology workflows employing robotized cell culture, sample preparation and data acquisition. In this setting, fluorescent biosensor cells that express microtubule-associated proteins 1A/1B light chain 3B (best known as LC3) conjugated to green fluorescent protein (GFP), are utilized together with automated high content microscopy for the image-based assessment of autophagy. In sum, we detail a drug discovery screening workflow from high throughput sample preparation and processing to data acquisition and analysis.

Published
2021

40. Live cell imaging of LC3 dynamics

Author

Giulia, Cerrato, Allan, Sauvat, Oliver, Kepp, and Guido, Kroemer

Subjects
Cell Nucleus, Cytoplasm, Green Fluorescent Proteins, Autophagosomes, Autophagy, Microtubule-Associated Proteins
Abstract

Macroautophagy (hereafter referred to as autophagy) serves the liberation of energy resources through the degradation of cellular components and is characterized by the formation of double-membraned vesicles, commonly referred to as autophagosomes. Microtubule-associated proteins 1A/1B light chain 3B (hereafter referred to as LC3) plays a crucial role during autophagosome formation, as cleavage of its immature form and subsequent conjugation to phosphatidylethanolamine facilitates autophagosomal membrane biogenesis. Indeed, the redistribution of green fluorescent protein (GFP)-conjugated LC3 from a diffuse cytosolic pattern into forming autophagosomes constitutes a morphological phenotype (commonly referred to as LC3 puncta) applicable to phenotypic analysis. The quantification of LC3 puncta in end-point assays has extensively been used in the past, allowing for the identification of autophagy modulators. Here, we describe a robust method employing automated confocal live cell imaging for the study of time-resolved LC3 dynamics. Furthermore, this method can be used to differentiate between phenotypes such as the homogeneous distribution of LC3 puncta in the cytoplasm, and the aggregation of LC3 clusters juxtaposed to the nucleus thus allowing for functional predictions.

Published
2021

41. ColocalizR: An open-source application for cell-based high-throughput colocalization analysis

Author

Marion Leduc, Guido Kroemer, Oliver Kepp, Allan Sauvat, Kevin Müller, Université Paris Descartes - Paris 5 (UPD5), Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), and Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université de Paris (UP)

Subjects
0301 basic medicine, Source code, Overlap coefficient, [SDV]Life Sciences [q-bio], Systems biology, media_common.quotation_subject, Intracellular Space, Health Informatics, Rendering (computer graphics), User-Computer Interface, 03 medical and health sciences, symbols.namesake, 0302 clinical medicine, Cell Line, Tumor, Image Processing, Computer-Assisted, Humans, media_common, Graphical user interface, business.industry, Systems Biology, Colocalization, Pattern recognition, Pearson product-moment correlation coefficient, Computer Science Applications, 030104 developmental biology, Microscopy, Fluorescence, symbols, Artificial intelligence, business, Software, 030217 neurology & neurosurgery, Cell based
Abstract

The microscopic assessment of the colocalization of fluorescent signals has been widely used in cell biology. Although imaging techniques have drastically improved over the past decades, the quantification of colocalization by measures such as the Pearson correlation coefficient or Manders overlap coefficient, has not changed. Here, we report the development of an R-based application that allows to (i) automatically segment cells and subcellular compartments, (ii) measure morphology and texture features, and (iii) calculate the degree of colocalization within each cell. Colocalization can thus be studied on a cell-by-cell basis, permitting to perform statistical analyses of cellular populations and subpopulations. ColocalizR has been designed to parallelize tasks, making it applicable to the analysis of large data sets. Its graphical user interface makes it suitable for researchers without specific knowledge in image analysis. Moreover, results can be exported into a wide range of formats rendering post-analysis adaptable to statistical requirements. This application and its source code are freely available at https://github.com/kroemerlab/ColocalizR.

Published
2019
Full Text
View/download PDF

42. Metabolomic analyses of COVID-19 patients unravel stage-dependent and prognostic biomarkers

Author

Giulia Baciarello, Guido Kroemer, Fanny Pommeret, Allan Sauvat, Eric Solary, Delphine Bredel, Nitharsshini Nirmalathasan, Agathe Dubuisson, Lisa Derosa, Annabelle Stoclin, Frank Griscelli, Mathilde Roumier, Fabrice Andre, Jean-Marie Michot, Frédéric Pène, Claudia Grajeda-Iglesias, Jean-Charles Soria, François-Xavier Danlos, Fabrice Barlesi, Jérôme Duchemin, Flore Rozenberg, Caroline Pradon, Françoise Levavasseur, Anne-Gaëlle Goubet, Julien Rohmer, Luc Mouthon, Laurence Zitvogel, Laurence Albiges, Severine Mouraud, Jean-Eudes Fahrner, Christophe Willekens, Sylvère Durand, Emeline Colomba, Aurélien Marabelle, Félix Ackerman, Syrine Ellouze, Michaela Fontenay, Georges Jourdi, Marc Vasse, Delphine Cantin, Immunologie anti-tumorale et immunothérapie des cancers (ITIC), Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay, Institut Gustave Roussy (IGR), Immunologie des tumeurs et immunothérapie (UMR 1015), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPCité), Institut Universitaire de France (IUF), Ministère de l'Education nationale, de l’Enseignement supérieur et de la Recherche (M.E.N.E.S.R.), Métabolisme, Cancer et Immunité (CRC - UMR_S 1138), Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPCité)-Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPCité)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Service de Médecine Interne [Hôpital Foch, Suresnes] (SMI), Hôpital Foch [Suresnes], Oncologie gynécologique, Département de médecine oncologique [Gustave Roussy], Institut Gustave Roussy (IGR)-Institut Gustave Roussy (IGR), Centre de Référence National des Syndromes Hyperéosinophiliques (CEREO), Département de cancérologie de l'enfant et de l'adolescent [Gustave Roussy], Hémostase, Inflammation, Thrombose (HITH - U1176 Inserm - CHU Bicêtre), Institut National de la Santé et de la Recherche Médicale (INSERM)-AP-HP Hôpital Bicêtre (Le Kremlin-Bicêtre)-Université Paris-Saclay, Université Paris-Saclay, Département de biologie et pathologie médicales [Gustave Roussy], Département de soins aigus [Gustave Roussy] (DSA), Direction de la recherche [Gustave Roussy], ANR-16-RHUS-0008,LUMIERE,LUMIERE(2016), ANR-18-IDEX-0001,Université de Paris,Université de Paris(2018), and European Project: 825410,ONCOBIOME

Subjects
Male, Cancer Research, Kynurenine pathway, Metabolite, [SDV]Life Sciences [q-bio], Immunology, [SDV.CAN]Life Sciences [q-bio]/Cancer, Disease, Pharmacology, Antibodies, Monoclonal, Humanized, Article, Cellular and Molecular Neuroscience, chemistry.chemical_compound, Metabolomics, Tocilizumab, Intensive care, medicine, Metabolome, Humans, lcsh:QH573-671, Pneumonitis, SARS-CoV-2, business.industry, lcsh:Cytology, COVID-19, Cell Biology, Prognosis, medicine.disease, COVID-19 Drug Treatment, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, chemistry, Viral infection, Female, business, Biomarkers
Abstract

The circulating metabolome provides a snapshot of the physiological state of the organism responding to pathogenic challenges. Here we report alterations in the plasma metabolome reflecting the clinical presentation of COVID-19 patients with mild (ambulatory) diseases, moderate disease (radiologically confirmed pneumonitis, hospitalization and oxygen therapy), and critical disease (in intensive care). This analysis revealed major disease- and stage-associated shifts in the metabolome, meaning that at least 77 metabolites including amino acids, lipids, polyamines and sugars, as well as their derivatives, were altered in critical COVID-19 patient’s plasma as compared to mild COVID-19 patients. Among a uniformly moderate cohort of patients who received tocilizumab, only 10 metabolites were different among individuals with a favorable evolution as compared to those who required transfer into the intensive care unit. The elevation of one single metabolite, anthranilic acid, had a poor prognostic value, correlating with the maintenance of high interleukin-10 and -18 levels. Given that products of the kynurenine pathway including anthranilic acid have immunosuppressive properties, we speculate on the therapeutic utility to inhibit the rate-limiting enzymes of this pathway including indoleamine 2,3-dioxygenase and tryptophan 2,3-dioxygenase.

Published
2021
Full Text
View/download PDF

43. A TLR3 Ligand Reestablishes Chemotherapeutic Responses in the Context of FPR1 Deficiency

Author

Francesca Castoldi, Maria Chiara Maiuri, Julie Le Naour, Julien Taieb, Allan Sauvat, Suzette Delaloge, Charles-Antoine Dutertre, Jessica Zucman-Rossi, Juliette Paillet, Erika Vacchelli, Liwei Zhao, Giulia Cerrato, Guido Kroemer, Claire Mulot, Isabelle Martins, Oliver Kepp, Federico Pietrocola, Pierre Laurent-Puig, Aymeric Silvin, Florent Ginhoux, Fabrice Andre, Peng Liu, Laurence Zitvogel, Sandy Adjemian, Cornelia Richter, Zoltan Szallasi, Peter Vandenabeele, Zsofia Sztupinszki, Jonathan Pol, Gautier Stoll, Khady Mangane, Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPCité), Institut Gustave Roussy (IGR), Institut Universitaire de France (IUF), Ministère de l'Education nationale, de l’Enseignement supérieur et de la Recherche (M.E.N.E.S.R.), Universidade de São Paulo = University of São Paulo (USP), Boston Children's Hospital, Harvard Medical School [Boston] (HMS), Cancer Research and Personalized Medicine - CARPEM [Paris], Hôpital Européen Georges Pompidou [APHP] (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Immunologie des tumeurs et immunothérapie (UMR 1015), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Agency for science, technology and research [Singapore] (A*STAR), Singapore Immunology Network (SIgN), Biomedical Sciences Institute (BMSI), School of Medicine [Shanghai Jiaotong University], Shanghai Jiaotong University, Métabolisme, Cancer et Immunité (CRC - UMR_S 1138), Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPCité)-Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPCité)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), University of Oslo (UiO), Direction de la recherche [Gustave Roussy], Département de médecine oncologique [Gustave Roussy], Pathologie mammaire, Institut Gustave Roussy (IGR)-Institut Gustave Roussy (IGR), Immunologie anti-tumorale et immunothérapie des cancers (ITIC), Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay, ANR-16-RHUS-0008,LUMIERE,LUMIERE(2016), European Project: 825410,ONCOBIOME, Le Naour, Julie, Liu, Peng, Zhao, Liwei, Adjemian, Sandy, Sztupinszki, Zsofia, Taieb, Julien, Mulot, Claire, Silvin, Aymeric, Dutertre, Charles-Antoine, Ginhoux, Florent, Sauvat, Allan, Cerrato, Giulia, Castoldi, Francesca, Martins, Isabelle, Stoll, Gautier, Paillet, Juliette, Mangane, Khady, Richter, Cornelia, Kepp, Oliver, Maiuri, Maria Chiara, Pietrocola, Federico, Vandenabeele, Peter, André, Fabrice, Delaloge, Suzette, Szallasi, Zoltan, Laurent-Puig, Pierre, Zucman-Rossi, Jessica, Zitvogel, Laurence, Pol, Jonathan G, Vacchelli, Erika, Kroemer, Guido, École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPC), Universidade de São Paulo (USP), Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPC)-Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPC)-École pratique des hautes études (EPHE), and Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPC)

Subjects
0301 basic medicine, Anthracycline, medicine.medical_treatment, [SDV]Life Sciences [q-bio], Mice, Transgenic, Ligands, 03 medical and health sciences, Mice, 0302 clinical medicine, Immune system, Medicine, Animals, Humans, ComputingMilieux_MISCELLANEOUS, business.industry, Pattern recognition receptor, Wild type, Immunotherapy, Receptors, Formyl Peptide, Toll-Like Receptor 3, Disease Models, Animal, 030104 developmental biology, Cell Transformation, Neoplastic, Poly I-C, Oncology, 030220 oncology & carcinogenesis, Cancer cell, TLR3, Cancer research, business, Colorectal Neoplasms, Annexin A1
Abstract

For anthracycline-based chemotherapy to be immunogenic, dying cancer cells must release annexin A1 (ANXA1) that subsequently interacts with the pattern recognition receptor, formyl peptide receptor 1 (FPR1), on the surface of dendritic cells (DC). Approximately 30% of individuals bear loss-of-function alleles of FPR1, calling for strategies to ameliorate their anticancer immune response. Here, we show that immunotherapy with a ligand of Toll-like receptor-3, polyinosinic:polycytidylic acid (pIC), restores the deficient response to chemotherapy of tumors lacking ANXA1 developing in immunocompetent mice or those of normal cancers growing in FPR1-deficient mice. This effect was accompanied by improved DC- and T-lymphocyte–mediated anticancer immunity. Of note, carcinogen-induced breast cancers precociously developed in FPR1-deficient mice as compared with wild-type controls. A similar tendency for earlier cancer development was found in patients carrying the loss-of-function allele of FPR1. These findings have potential implications for the clinical management of FPR1-deficient patients. Significance: The loss-of-function variant rs867228 in FPR1, harbored by approximately 30% of the world population, is associated with the precocious manifestation of breast, colorectal, esophageal, and head and neck carcinomas. pIC restores deficient chemotherapeutic responses in mice lacking Fpr1, suggesting a personalized strategy for compensating for the FPR1 defect. This article is highlighted in the In This Issue feature, p. 211

Published
2021
Full Text
View/download PDF

44. High throughput screening for autophagy

Author

Giulia Cerrato, Sabrina Forveille, Allan Sauvat, Marion Leduc, Guido Kroemer, and Oliver Kepp

Subjects
Drug discovery, High-throughput screening, Autophagy, Computational biology, Biology, Throughput (business), Biosensor, Green fluorescent protein
Abstract

Robotized high throughput screening allows for the assessment of autophagy in a large number of samples. Here, we describe a drug discovery platform for the phenotypic identification of novel autophagy inducers by means of automated cell biology workflows employing robotized cell culture, sample preparation and data acquisition. In this setting, fluorescent biosensor cells that express microtubule-associated proteins 1A/1B light chain 3B (best known as LC3) conjugated to green fluorescent protein (GFP), are utilized together with automated high content microscopy for the image-based assessment of autophagy. In sum, we detail a drug discovery screening workflow from high throughput sample preparation and processing to data acquisition and analysis.

Published
2021
Full Text
View/download PDF

45. Clonogenic Assays to Detect Cell Fate in Mitotic Catastrophe

Author

Laura Senovilla, José Manuel Bravo-San Pedro, Allan Sauvat, Guido Kroemer, Santiago Rello-Varona, and Oliver Kepp

Subjects
0301 basic medicine, Cell cycle checkpoint, DNA damage, Cell, Biology, Cell biology, 03 medical and health sciences, Spindle checkpoint, 030104 developmental biology, 0302 clinical medicine, medicine.anatomical_structure, Centrosome, 030220 oncology & carcinogenesis, medicine, Clonogenic assay, Mitosis, Mitotic catastrophe
Abstract

Mitotic catastrophe (MC) is a cell death modality induced by DNA damage that involves the activation of cell cycle checkpoints such as the "DNA structure checkpoint" and "spindle assembly checkpoint" (SAC) leading to aberrant mitosis. Depending on the signal, MC can drive the cell to death or to senescence. The suppression of MC favors aneuploidy. Several cancer therapies, included microtubular poisons and radiations, trigger MC. The clonogenic assay has been used to study the capacity of single cells to proliferate and to generate macroscopic colonies and to evaluate the efficacy of anticancer drugs. Nevertheless, this method cannot analyze MC events. Here, we report an improved technique based on the use of human colon cancer HCT116 stable expressing histone H2B-GFP and DsRed-centrin proteins, allowing to determine the capacity of cells to proliferate, and to determine changes in the nucleus and centrosomes.

Published
2021
Full Text
View/download PDF

46. Live cell imaging of LC3 dynamics

Author

Giulia Cerrato, Guido Kroemer, Oliver Kepp, and Allan Sauvat

Subjects
Vesicle, Autophagy, Biology, Cell biology, Green fluorescent protein, Cytosol, medicine.anatomical_structure, Cytoplasm, Live cell imaging, embryonic structures, Membrane biogenesis, medicine, biological phenomena, cell phenomena, and immunity, Nucleus
Abstract

Macroautophagy (hereafter referred to as autophagy) serves the liberation of energy resources through the degradation of cellular components and is characterized by the formation of double-membraned vesicles, commonly referred to as autophagosomes. Microtubule-associated proteins 1A/1B light chain 3B (hereafter referred to as LC3) plays a crucial role during autophagosome formation, as cleavage of its immature form and subsequent conjugation to phosphatidylethanolamine facilitates autophagosomal membrane biogenesis. Indeed, the redistribution of green fluorescent protein (GFP)-conjugated LC3 from a diffuse cytosolic pattern into forming autophagosomes constitutes a morphological phenotype (commonly referred to as LC3 puncta) applicable to phenotypic analysis. The quantification of LC3 puncta in end-point assays has extensively been used in the past, allowing for the identification of autophagy modulators. Here, we describe a robust method employing automated confocal live cell imaging for the study of time-resolved LC3 dynamics. Furthermore, this method can be used to differentiate between phenotypes such as the homogeneous distribution of LC3 puncta in the cytoplasm, and the aggregation of LC3 clusters juxtaposed to the nucleus thus allowing for functional predictions.

Published
2021
Full Text
View/download PDF

47. Local anesthetics elicit immune-dependent anticancer effects

Author

Lucillia Bezu, Alejandra Wu Chuang, Allan Sauvat, Juliette Humeau, Wei Xie, Giulia Cerrato, Peng Liu, Liwei Zhao, Shuai Zhang, Julie Le Naour, Jonathan Pol, Peter van Endert, Oliver Kepp, Fabrice Barlesi, Guido Kroemer, Métabolisme, cancer et immunité = Metabolism, Cancer & Immunity [CRC] (Equipe labellisée Ligue contre le cancer), Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPCité)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPCité), Institut Gustave Roussy (IGR), Institut de Recherche en Immunologie et en Cancérologie [UdeM-Montréal] (IRIC), Université de Montréal (UdeM), Institut Universitaire de France (IUF), Ministère de l'Education nationale, de l’Enseignement supérieur et de la Recherche (M.E.N.E.S.R.), Huazhong University of Science and Technology [Wuhan] (HUST), Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Descartes - Paris 5 (UPD5), Sorbonne Université - École supérieure du professorat et de l'éducation - Académie de Paris (ESPE Paris), Sorbonne Université (SU), Apoptose, cancer et immunité (Equipe labellisée Ligue contre le cancer - CRC - Inserm U1138), Institut Gustave Roussy (IGR)-Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Département de médecine oncologique [Gustave Roussy], Centre de Recherche en Cancérologie de Marseille (CRCM), Aix Marseille Université (AMU)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Cancer Research and Personalized Medicine - CARPEM [Paris], Hôpital Européen Georges Pompidou [APHP] (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Necker - Enfants Malades [AP-HP], and Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)

Subjects
Cancer Research, STRESS, [SDV]Life Sciences [q-bio], Cytotoxicity, Eukaryotic Initiation Factor-2, Immunology, PERIPHERAL-BLOOD, Endoplasmic Reticulum, LIDOCAINE, NATURAL-KILLER-CELLS, Mice, Drug Therapy, Immunologic, NEURAXIAL ANESTHESIA, Neoplasms, Medicine and Health Sciences, Animals, Humans, Immunology and Allergy, Adjuvants, Anesthetics, Local, RECURRENCE, Retrospective Studies, Pharmacology, CALRETICULIN EXPOSURE, Biology and Life Sciences, Activating Transcription Factor 4, Preclinical, CANCER-SURGERY, Oncology, Combination, SURVIVAL, Drug Evaluation, Molecular Medicine, AUTOPHAGY, Immunotherapy
Abstract

BackgroundRetrospective clinical trials reported a reduced local relapse rate, as well as improved overall survival after injection of local anesthetics during cancer surgery. Here, we investigated the anticancer effects of six local anesthetics used in clinical practice.ResultsIn vitro, local anesthetics induced signs of cancer cell stress including inhibition of oxidative phosphorylation, and induction of autophagy as well as endoplasmic reticulum (ER) stress characterized by the splicing of X-box binding protein 1 (XBP1s) mRNA, cleavage of activating transcription factor 6 (ATF6), phosphorylation of eIF2α and subsequent upregulation of activating transcription factor 4 (ATF4). Both eIF2α phosphorylation and autophagy required the ER stress-relevant eukaryotic translation initiation factor 2 alpha kinase 3 (EIF2AK3, best known as PERK). Local anesthetics also activated two hallmarks of immunogenic cell death, namely, the release of ATP and high-mobility group box 1 protein (HMGB1), yet failed to cause the translocation of calreticulin (CALR) from the ER to the plasma membrane. In vivo, locally injected anesthetics decreased tumor growth and improved survival in several models of tumors established in immunocompetent mice. Systemic immunotherapy with PD-1 blockade or intratumoral injection of recombinant CALR protein, increased the antitumor effects of local anesthetics. Local anesthetics failed to induce antitumor effects in immunodeficient mice or against cancers unable to activate ER stress or autophagy due to the knockout of EIF2AK3/PERK or ATG5, respectively. Uncoupling agents that inhibit oxidative phosphorylation and induce autophagy and ER stress mimicked the immune-dependent antitumor effects of local anesthetics.ConclusionAltogether, these results indicate that local anesthetics induce a therapeutically relevant pattern of immunogenic stress responses in cancer cells.

Published
2022
Full Text
View/download PDF

48. A genome-wide RNA interference screen disentangles the Golgi tropism of LC3

Author

Allan Sauvat, Giulia Cerrato, Guido Kroemer, and Oliver Kepp

Subjects
education.field_of_study, Endoplasmic reticulum, Golgi Apparatus, Cell Biology, Biology, Golgi apparatus, Tropism, Article, Cell biology, Transport protein, Eukaryotic translation initiation factor 2A, symbols.namesake, Protein Transport, Secretory protein, RNA interference, Unfolded protein response, symbols, Autophagy, RNA Interference, education, Molecular Biology, MAP1LC3B, Oleic Acid, trans-Golgi Network
Abstract

Oleate, the most abundantly occurring cis-unsaturated fatty acid, has the particularity to induce the accumulation of MAP1LC3B/LC3 (microtubule associated protein 1 light chain 3 beta) at the trans-Golgi apparatus. A genome-wide RNA interference screen designed to identify the mechanisms of this LC3 redistribution led to the identification of a BECN1-PIK3C3-independent pathway that, however, requires the ATG12-ATG5 and ATG7-dependent conjugation system, and several genes/proteins involved in endoplasmic reticulum (ER)-to-Golgi anterograde protein transport, as well as the unfolded protein response, including the integrated stress response that results in the phosphorylation of EIF2A/eIF2α (eukaryotic translation initiation factor 2A). Functional experiments revealed that oleate blocks conventional protein secretion, stalling the process at the level of the trans-Golgi network. Oleate-induced blockade of protein secretion occurred even after depletion of ATG5, suggesting that it does not rely on the recruitment of LC3 to the Golgi apparatus (which does require ATG5). Rather, it appears that oleate and other pharmacological inhibitors of protein secretion with a similar mode of action provoke a perturbation of the trans-Golgi compartment that secondarily results in the local enrichment of LC3.

Published
2020

49. TiO2 Nanomaterials Non-Controlled Contamination Could Be Hazardous for Normal Cells Located in the Field of Radiotherapy

Author

Celine Lacrouts, Fabrice Nesslany, Romain Grall, Sylvie Chevillard, Jérôme Lebeau, Yidan Wang, Marie Hullo, Allan Sauvat, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Institut Gustave Roussy (IGR), Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPCité), Impact de l'environnement chimique sur la santé humaine - ULR 4483 (IMPECS), Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), CHU Lille, Institut Pasteur de Lille, Université de Lille, Commissariat à l'énergie atomique et aux énergies alternatives [CEA], Institut Gustave Roussy [IGR], and IMPact de l'Environnement Chimique sur la Santé humaine (IMPECS) - ULR 4483

Subjects
TiO2 nanoparticles, 0301 basic medicine, UVA Radiation, Cell Survival, medicine.medical_treatment, Metal Nanoparticles, 02 engineering and technology, [SDV.IB.MN]Life Sciences [q-bio]/Bioengineering/Nuclear medicine, Article, Catalysis, Cell Line, Ionizing radiation, lcsh:Chemistry, Inorganic Chemistry, 03 medical and health sciences, Radiation sensitivity, medicine, Humans, Physical and Theoretical Chemistry, lcsh:QH301-705.5, Bronchioles, Molecular Biology, photocatalysis, radiation sensitivity, Spectroscopy, Cell Proliferation, Titanium, Skin care, Radiotherapy, Chemistry, Organic Chemistry, technology, industry, and agriculture, General Medicine, Contamination, 021001 nanoscience & nanotechnology, Computer Science Applications, Radiation exposure, Radiation therapy, Oxidative Stress, 030104 developmental biology, lcsh:Biology (General), lcsh:QD1-999, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Cancer research, Cell response, Reactive Oxygen Species, 0210 nano-technology
Abstract

Among nanomaterials (NMs), titanium dioxide (TiO2) is one of the most manufactured NMs and can be found in many consumers&rsquo, products such as skin care products, textiles and food (as E171 additive). Moreover, due to its most attractive property, a photoactivation upon non-ionizing UVA radiation, TiO2 NMs is widely used as a decontaminating agent. Uncontrolled contaminations by TiO2 NMs during their production (professional exposure) or by using products (consumer exposure) are rather frequent. So far, TiO2 NMs cytotoxicity is still a matter of controversy depending on biological models, types of TiO2 NMs, suspension preparation and biological endpoints. TiO2 NMs photoactivation has been widely described for UV light radiation exposure, it could lead to reactive oxygen species production, known to be both cyto- and genotoxic on human cells. After higher photon energy exposition, such as X-rays used for radiotherapy and for medical imaging, TiO2 NMs photoactivation still occurs. Importantly, the question of its hazard in the case of body contamination of persons receiving radiotherapy was never addressed, knowing that healthy tissues surrounding the tumor are indeed exposed. The present work focuses on the analysis of human normal bronchiolar cell response after co-exposition TiO2 NMs (with different coatings) and ionizing radiation. Our results show a clear synergistic effect, in terms of cell viability, cell death and oxidative stress, between TiO2 NMS and radiation.

Published
2020
Full Text
View/download PDF

50. An unexpected link between immunogenic cell death and inhibition of gene transcription

Author

Allan Sauvat, Juliette Humeau, Guido Kroemer, and Oliver Kepp

Subjects
0301 basic medicine, Transcription, Genetic, Immunology, Apoptosis, Immunogenic Cell Death, Biology, 03 medical and health sciences, 0302 clinical medicine, Transcription (biology), Neoplasms, medicine, Humans, Immunology and Allergy, Malignant cells, RC254-282, Cancer, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC581-607, medicine.disease, Editorial, 030104 developmental biology, Oncology, 030220 oncology & carcinogenesis, Cancer research, Immunogenic cell death, Immunologic diseases. Allergy
Abstract

Colliding with the ancient concept that chemotherapeutics may delay (and sometimes stop or even reverse) the advancement of cancer because they kill malignant cells, it has become clear that succes...

Published
2020

Catalog

Books, media, physical & digital resources
See catalog results