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150 results on '"Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS)"'

1. Interdisciplinary strategy to assess the impact of meteorological variables on the biochemical composition of the rain and the dynamics of a small eutrophic lake under rain forcing

Author: Fanny Noirmain, Jean-Luc Baray, Frédéric Tridon, Philippe Cacault, Hermine Billard, Guillaume Voyard, Joël Van Baelen, Delphine Latour, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Laboratoire de Météorologie Physique (LaMP), Institut national des sciences de l'Univers (INSU - CNRS)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Centre National de la Recherche Scientifique (CNRS), Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Clermont Auvergne (UCA), Observatoire de Physique du Globe de Clermont-Ferrand (OPGC), Laboratoire Microorganismes : Génome et Environnement (LMGE), Centre National de la Recherche Scientifique (CNRS)-Université Clermont Auvergne (UCA), University of California [Berkeley] (UC Berkeley), University of California (UC), Université Clermont Auvergne (UCA), Institut de Chimie de Clermont-Ferrand (ICCF), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Clermont Auvergne (UCA)-Institut national polytechnique Clermont Auvergne (INP Clermont Auvergne), Université Clermont Auvergne (UCA)-Université Clermont Auvergne (UCA), Laboratoire de l'Atmosphère et des Cyclones (LACy), and Institut national des sciences de l'Univers (INSU - CNRS)-Université de La Réunion (UR)-Centre National de la Recherche Scientifique (CNRS)-Météo-France
Subjects: impact of meteorological variables, [SDE]Environmental Sciences, biochemical composition of rain, dynamics of a small eutrophic lake under rain forcing, Ecology, Evolution, Behavior and Systematics, Earth-Surface Processes
Abstract: We present an interdisciplinary investigation of the links between the macro and microphysical properties of rain, the biochemical composition of rain, meteorological parameters, and their impacts on the phytoplankton dynamics of a mountain lake. In order to document this interdisciplinary scientific question, Lake Aydat in the French Massif Central mountains was fitted with a set of high-resolution atmospheric radars, a disdrometer, and a precipitation collector in 2020. In parallel, the lake was monitored via sensors and water sampling. To illustrate the potential of this novel experimental setup, we present a case study of a rain event that occurred in September 2020 and during which, three contrasting sub-periods were identified based on the type of rain (convective or stratiform). Using our high-temporal resolution monitoring, we show that the origin of the air mass mainly influences the chemical composition of the rain, which depends on the rain type. In contrast, the photosynthetic cell concentration in the rain is mainly influenced by meteorological variables, predominantly below-cloud scavenging. The very low concentrations of photosynthetic cells found in rain events cannot directly impact the lake's phytoplankton abundance. In contrast, the rain rate directly impacted the lake's thermal stratification during the convective event. The response of the phytoplankton depends on the genus and, interestingly, three cyanobacteria, Microcystis, Coelomoron, and Merismopedia, showed a systematic response to rain events with a sudden decrease in abundance at the lake surface immediately after rain events. In contrast, the abundance of green algae (Elakatothrix), picocyanobacteria (Synechocystis and Synechococcus), and diatoms (Asterionella and Melosira) gradually increased following the rain events, but with a lower intensity compared to the cyanobacteria species. These different phytoplankton responses to the same rainfall event could play a key role in phytoplankton dynamics in the temperate zone. Our results highlight the importance of high-frequency and time resolution monitoring of both atmosphere and lake to better understand the adaptive strategies of cyanobacteria following rain events.
Published: 2022
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2. Matrix Metalloproteinases: From Molecular Mechanisms to Physiology, Pathophysiology, and Pharmacology

Author: Luiz G.N. de Almeida, Hayley Thode, Yekta Eslambolchi, Sameeksha Chopra, Daniel Young, Sean Gill, Laurent Devel, Antoine Dufour, University of Calgary, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
Subjects: Pharmacology, Neoplasms, [SDV]Life Sciences [q-bio], Proteolysis, Humans, [CHIM]Chemical Sciences, Molecular Medicine, Matrix Metalloproteinase Inhibitors, Matrix Metalloproteinases, Extracellular Matrix
Abstract: International audience; The first matrix metalloproteinase (MMP) was discovered in 1962 from the tail of a tadpole by its ability to degrade collagen. As their name suggests, matrix metalloproteinases are proteases capable of remodeling the extracellular matrix. More recently, MMPs have been demonstrated to play numerous additional biologic roles in cell signaling, immune regulation, and transcriptional control, all of which are unrelated to the degradation of the extracellular matrix. In this review, we will present milestones and major discoveries of MMP research, including various clinical trials for the use of MMP inhibitors. We will discuss the reasons behind the failures of most MMP inhibitors for the treatment of cancer and inflammatory diseases. There are still misconceptions about the pathophysiological roles of MMPs and the best strategies to inhibit their detrimental functions. This review aims to discuss MMPs in preclinical models and human pathologies. We will discuss new biochemical tools to track their proteolytic activity in vivo and ex vivo, in addition to future pharmacological alternatives to inhibit their detrimental functions in diseases. SIGNIFICANCE STATEMENT: Matrix metalloproteinases (MMPs) have been implicated in most inflammatory, autoimmune, cancers, and pathogen-mediated diseases. Initially overlooked, MMP contributions can be both beneficial and detrimental in disease progression and resolution. Thousands of MMP substrates have been suggested, and a few hundred have been validated. After more than 60 years of MMP research, there remain intriguing enigmas to solve regarding their biological functions in diseases.Significance Statement⎯MMPs have been implicated in most inflammatory, autoimmune, cancers and pathogen-mediated diseases. Initially overlooked, MMP contributions can be both beneficial and detrimental in disease progression and resolution. Thousands of MMP substrates have been suggested and a few hundred have been validated. After more than 60 years of MMP research, there remain intriguing enigmas to solve regarding their biological functions in diseases.
Published: 2022
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3. Deciphering cross-species reactivity of LAMP-1 antibodies using deep mutational epitope mapping and AlphaFold

Author: Tiphanie Pruvost, Magali Mathieu, Steven Dubois, Bernard Maillère, Emmanuelle Vigne, Hervé Nozach, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Sanofi [Vitry-sur-Seine], and SANOFI Recherche
Subjects: epitope mapping, deep mutational scanning, yeast surface display, LAMP-1 DMS: Deep Mutational Scanning YSD: Yeast Surface Display NGS: next-generation sequencing cryo-EM: cryogenic electron microscopy BLI: biolayer interferometry FACS: Fluorescence-activated cell sorting CDRs: complementarity-determining regions HDX-MS: Hydrogen deuterium exchange mass spectrometry RMSD: root-mean-square deviation, [SDV]Life Sciences [q-bio], cross-species reactivity, Immunology, Immunology and Allergy, Monoclonal antibodies, Monoclonal antibodies deep mutational scanning yeast surface display epitope mapping cross-species reactivity LAMP-1 DMS: Deep Mutational Scanning YSD: Yeast Surface Display NGS: next-generation sequencing cryo-EM: cryogenic electron microscopy BLI: biolayer interferometry FACS: Fluorescence-activated cell sorting CDRs: complementarity-determining regions HDX-MS: Hydrogen deuterium exchange mass spectrometry RMSD: root-mean-square deviation
Abstract: International audience; Delineating the precise regions on an antigen that are targeted by antibodies has become a key step for the development of antibody therapeutics. X-ray crystallography and cryogenic electron microscopy are considered the gold standard for providing precise information about these binding sites at atomic resolution. However, they are labor-intensive and a successful outcome is not guaranteed. We used deep mutational scanning (DMS) of the human LAMP-1 antigen displayed on yeast surface and leveraged next-generation sequencing to observe the effect of individual mutants on the binding of two LAMP-1 antibodies and to determine their functional epitopes on LAMP-1. Fine-tuned epitope mapping by DMS approaches is augmented by knowledge of experimental antigen structure. As human LAMP-1 structure has not yet been solved, we used the AlphaFold predicted structure of the full-length protein to combine with DMS data and ultimately finely map antibody epitopes. The accuracy of this method was confirmed by comparing the results to the co-crystal structure of one of the two antibodies with a LAMP-1 luminal domain. Finally, we used AlphaFold models of non-human LAMP-1 to understand the lack of mAb cross-reactivity. While both epitopes in the murine form exhibit multiple mutations in comparison to human LAMP-1, only one and two mutations in the Macaca form suffice to hinder the recognition by mAb B and A, respectively. Altogether, this study promotes a new application of AlphaFold to speed up precision mapping of antibody–antigen interactions and consequently accelerate antibody engineering for optimization.
Published: 2023
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4. Editorial of the special issue 'Toxins: Mr Hyde or Dr Jekyll?'

Author: Daniel Ladant, Gilles Prévost, Michel R. Popoff, Evelyne Benoit, Biochimie des Interactions Macromoléculaires / Biochemistry of Macromolecular Interactions, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Virulence bactérienne précoce : fonctions cellulaires et contrôle de l'infection aiguë et subaiguë, Université de Strasbourg (UNISTRA), Toxines bactériennes - Bacterial Toxins, Institut Pasteur [Paris] (IP), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
Subjects: plant toxin, maine toxin, toxin structure, medical application, Health, Toxicology and Mutagenesis, [SDV.TOX]Life Sciences [q-bio]/Toxicology, bacterial toxin, animal toxin, Toxicology, toxin receptor, toxin function
Abstract: International audience; The 27th Annual Meeting of the French Society of Toxinology (SFET, http://sfet.asso.fr/international (accessed since 1 September 2022)) was held on 9–10 December 2021 as a virtual meeting (e-RT27). The central theme selected for this meeting, “Toxins: Mr Hyde or Dr Jekyll?”, gave rise to three thematic sessions: the first on plant toxins, algal toxins and mycotoxins; the second on animal toxins; and the third on bacterial toxins. All sessions were aimed at emphasizing the latest findings on this topic. Apart from this central theme, a “miscellaneous” session was dedicated to recent results obtained in toxinology. Ten speakers from seven countries (Australia, Brazil, Burkina Faso, France, Germany, The Netherlands and the United States of America) were invited as international experts to present their work, and other researchers and students presented theirs through 15 shorter lectures and 20 posters. Of the ca. 80 participants who registered, 38% were foreigners, a value highlighting the international attractiveness of the SFET meeting.
Published: 2023
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5. Sec22b is a critical and non-redundant regulator of plasma cell maintenance

Author: Amélie Bonaud, Laetitia Gargowitsch, Simon M. Gilbert, Elanchezhian Rajan, Pablo Canales-Herrerias, Daniel Stockholm, Nabila F. Rahman, Mark O. Collins, Hakan Taskiran, Danika L. Hill, Andres Alloatti, Nagham Alouche, Stéphanie Balor, Vanessa Soldan, Daniel Gillet, Julien Barbier, Françoise Bachelerie, Kenneth G. C. Smith, Julia Jellusova, Pierre Bruhns, Sebastian Amigorena, Karl Balabanian, Michelle A. Linterman, Andrew A. Peden, Marion Espéli, Institut de Recherche Saint-Louis - Hématologie Immunologie Oncologie (Département de recherche de l’UFR de médecine, ex- Institut Universitaire Hématologie-IUH) (IRSL), Université Paris Cité (UPCité), Microenvironnement des niches tumorales (CNRS GDR 3697 Micronit ), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Hopital Saint-Louis [AP-HP] (AP-HP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Inflammation, microbiome, immunosurveillance (MI2), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay, University of Cambridge [UK] (CAM), University of Sheffield [Sheffield], Anticorps en thérapie et pathologie - Antibodies in Therapy and Pathology, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL), 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), School of Psychology [Cardiff University], Cardiff University, Technische Universität München = Technical University of Munich (TUM), Max Planck Institute of Immunobiology and Epigenetics (MPI-IE), Max-Planck-Gesellschaft, The Babraham Institute [Cambridge, UK], Monash University [Melbourne], Immunité et cancer (U932), Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Universidad Nacional de Rosario [Santa Fe], Centre de Biologie Intégrative (CBI), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), ANR-19-CE15-0019,PC-SEC,Impact des SNARE sur la biologie des plasmocytes(2019), ANR-14-ACHN-0008,AUTO-PLASMO,Analyse integrative de la biologie des plasmocytes normaux et pathologiques(2014), ANR-17-CE14-0019,OSTEOVALYMPH,Autocrinie et paracrinie de l'axe de signalisation CXCL12/CXCR4-CXCR7 dans la niche ostéo-vasculaire: impact sur la spécification et l'engagement lymphoïde des cellules souches hématopoïétiques(2017), ANR-18-CE15-0001,Autoimmuni-B,Etude de la rupture de tolérance dans une maladie humaine autoimmune médiée par les lymphocytes B(2018), The study was supported by the Laboratory of Excellence in Research on Medication and Innovative Therapeutics (LabEx LERMIT) (M.E. and K.B.), an ANR JCJC grant (ANR-19-CE15-0019-01), an ANR @RAction grant (ANR-14-ACHN-0008), a 'Fondation ARC pour la recherche sur le cancer' grant (P JA20181208173), and a grant from IdEx Université Paris-Cité (ANR-18-IDEX-0001) to M.E., and an ANR PRC grant (ANR-17-CE14-0019) and an INCa grant (PRT-K 2017) to K.B. P.B. acknowledges funding from the French National Research Agency grant ANR-18-CE15-0001 project Autoimmuni-B, by the Institut Carnot Pasteur Microbes et Santé grant ANR-11-CARN-0017-01, the Institut Pasteur, and the Institut National de la Santé et de la Recherche Médicale (INSERM). M.A.L. is supported by Biotechnology and Biological Sciences Research Council (BBS/E/B/000C0427, BBS/E/B/000C0428, and the Campus Capability Core Grant to the Babraham Institute). A.A.P. and M.O.C. were supported by a grant from the Biotechnology and Biological Sciences Research Council (BB/L022389/1). D.L.H. is supported by a National Health and Medical Research Council Australia Early-Career Fellowship (APP1139911). N.A. was supported by a PhD fellowship from the French Ministry for education and by a fourth year PhD fellowship from the 'Fondation ARC pour la recherche sur le cancer.' P.C.-H. was supported partly by a stipend from the Pasteur-Paris University (PPU) International PhD program, and by a fellowship from the French Fondation pour la Recherche Médicale (FRM). K.G.C.S. was supported by the Wellcome Trust (Programme Grant Number 083650/Z/07/Z). J.J’s research is supported by the German Research Foundation project number: 419193696 and through the CRC1335. H.T. is supported through the graduate school of the Max Planck Institute for Immunobiology and Epigenetics (IMPRS-IE) and through the CRC1335. The 'EMiLy' U1160 INSERM unit is a member of the OPALE Carnot institute, The Organization for Partnerships in Leukemia (Institut Carnot OPALE, Institut de Recherche Saint-Louis, Hôpital Saint-Louis, Paris, France. Web: www.opale.org. Email: contact@opale.org)., ANR-18-IDEX-0001,Université de Paris,Université de Paris(2018), Bonaud, Amélie [0000-0002-4153-9171], Rajan, Elanchezhian [0000-0002-6257-3678], Canales-Herrerias, Pablo [0000-0002-1865-6476], Stockholm, Daniel [0000-0002-5069-5256], Collins, Mark O [0000-0002-7656-4975], Taskiran, Hakan [0000-0002-2690-3887], Alloatti, Andres [0000-0003-0555-0653], Gillet, Daniel [0000-0003-0477-3599], Bruhns, Pierre [0000-0002-4709-8936], Balabanian, Karl [0000-0002-0534-3198], Linterman, Michelle A [0000-0001-6047-1996], Peden, Andrew A [0000-0003-0144-7712], Espéli, Marion [0000-0001-5005-1664], and Apollo - University of Cambridge Repository
Subjects: R-SNARE Proteins, mitochondria, Mice, endoplasmic reticulum, Multidisciplinary, plasma cell, SNARE, antibody, [SDV]Life Sciences [q-bio], Plasma Cells, Animals, Biological Transport, SNARE Proteins
Abstract: Despite the essential role of plasma cells in health and disease, the cellular mechanisms controlling their survival and secretory capacity are still poorly understood. Here, we identified the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) Sec22b as a unique and critical regulator of plasma cell maintenance and function. In the absence of Sec22b, plasma cells were hardly detectable and serum antibody titers were dramatically reduced. Accordingly, Sec22b -deficient mice fail to mount a protective immune response. At the mechanistic level, we demonstrated that Sec22b contributes to efficient antibody secretion and is a central regulator of plasma cell maintenance through the regulation of their transcriptional identity and of the morphology of the endoplasmic reticulum and mitochondria. Altogether, our results unveil an essential and nonredundant role for Sec22b as a regulator of plasma cell fitness and of the humoral immune response.
Published: 2023
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6. Mode of action of cyclic imine toxins: 20-methyl spirolide-G antagonizes rat neuronal α3β2 nicotinic acetylcholine receptor

Author: Gaudin, Amandine, Aráoz, Rómulo, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Labex Lermit (Detectneurotox CDE2017-001173-RD91), NRBC-E (Multitox H35), Interreg Atlantic Area (Alertoxnet EAPA_317/2016), and Band-Schmidt, C.J. and Rodríguez-Gómez, C.F. (Eds.)
Subjects: twoelectrodes voltage clamp electrophysiology (TEVC), Cyclic imine toxins, two-electrodes voltage clamp electrophysiology (TEVC), [SDE]Environmental Sciences, nicotinic acetylcholine receptors, Xenopus laevis oocytes, 20-methyl spirolide-G
Abstract: Cyclic imine toxins produced by marine dinoflagellate species move up fast the food chain through shellfish and constitute a potential threat for marine wildlife and public health because of their potent antagonistic activity against nicotinic acetylcholine receptors of muscle and neuronal types. Although not regulated, the lethality of 13-desmethyl spirolide-C does compare the lethality of saxitoxin in mouse bioassay. Besides, cyclic imine toxins are often found in shellfish destined for human consumption. Herein, the unveiling of the mode of action of cyclic imine toxins is reviewed. To illustrate the review, the electrophysiological characterization of the antagonistic capacity of 20-methyl spirolide-G on rat α3β2 nicotinic acetylcholine receptor is described.
Published: 2022
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7. MT9, a natural peptide from black mamba venom antagonizes the muscarinic type 2 receptor and reverses the M2R-agonist-induced relaxation in rat and human arteries

Author: Justyna Ciolek, Claude Zoukimian, Justine Dhot, Mélanie Burban, Mathilde Triquigneaux, Benjamin Lauzier, Christelle Guimbert, Didier Boturyn, Marine Ferron, Lidia Ciccone, Livia Tepshi, Enrico Stura, Pierre Legrand, Philippe Robin, Gilles Mourier, Béatrice Schaack, Imen Fellah, Guillaume Blanchet, Chantal Gauthier-Erfanian, Rémy Beroud, Denis Servent, Michel De Waard, Nicolas Gilles, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), unité de recherche de l'institut du thorax UMR1087 UMR6291 (ITX), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Nantes Université - UFR de Médecine et des Techniques Médicales (Nantes Univ - UFR MEDECINE), Nantes Université - pôle Santé, Nantes Université (Nantes Univ)-Nantes Université (Nantes Univ)-Nantes Université - pôle Santé, Nantes Université (Nantes Univ)-Nantes Université (Nantes Univ), Synchrotron SOLEIL (SSOLEIL), Centre National de la Recherche Scientifique (CNRS), Translational microbial Evolution and Engineering (TIMC-TrEE), Translational Innovation in Medicine and Complexity / Recherche Translationnelle et Innovation en Médecine et Complexité - UMR 5525 (TIMC ), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP ), Université Grenoble Alpes (UGA)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP ), Université Grenoble Alpes (UGA), Département de Chimie Moléculaire (DCM), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Thérapeutique Recombinante Expérimentale (TIMC-IMAG-TheREx ), and Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications Grenoble - UMR 5525 (TIMC-IMAG)
Subjects: Elapid Venoms, Pharmacology, Bioactive peptide mass spectrometry, Snake venom, Dendroaspis, M2 muscarinic receptor, Cholinergic Agents, Arteries, General Medicine, Receptors, Muscarinic, Rats, Peptide synthesis, Drug screening, [SDV.MHEP.CSC]Life Sciences [q-bio]/Human health and pathology/Cardiology and cardiovascular system, Peptide folding, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, Animals, Humans, Peptides, Toxins, Biological, Arterial pressure
Abstract: International audience; All five muscarinic receptors have important physiological roles. The endothelial M2 and M3 subtypes regulate arterial tone through direct coupling to Gq or Gi/o proteins. Yet, we lack selective pharmacological drugs to assess the respective contribution of muscarinic receptors to a given function. We used mamba snake venoms to identify a selective M2R ligand to investigate its contribution to arterial contractions. Using a bio-guided screening binding assay, we isolated MT9 from the black mamba venom, a three-finger toxin active on the M2R subtype. After sequencing and chemical synthesis of MT9, we characterized its structure by X-ray diffraction and determined its pharmacological characteristics by binding assays, functional tests, and ex vivo experiments on rat and human arteries. Although MT9 belongs to the three-finger fold toxins family, it is phylogenetically apart from the previously discovered muscarinic toxins, suggesting that two groups of peptides evolved independently and in a convergent way to target muscarinic receptors. The affinity of MT9 for the M2R is 100 times stronger than that for the four other muscarinic receptors. It also antagonizes the M2R/G i pathways in cell-based assays. MT9 acts as a non-competitive antagonist against acetylcholine or arecaine, with low nM potency, for the activation of isolated rat mesenteric arteries. These results were confirmed on human internal mammary arteries. In conclusion, MT9 is the first fully characterized M2R-specific natural toxin. It should provide a tool for further understanding of the effect of M2R in various arteries and may position itself as a new drug candidate in cardiovascular diseases
Published: 2022
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8. Deep mutational engineering of broadly-neutralizing nanobodies accommodating SARS-CoV-1 and 2 antigenic drift

Author: Adrien Laroche, Maria Lucia Orsini Delgado, Benjamin Chalopin, Philippe Cuniasse, Steven Dubois, Raphaël Sierocki, Fabrice Gallais, Stéphanie Debroas, Laurent Bellanger, Stéphanie Simon, Bernard Maillère, Hervé Nozach, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Service de Pharmacologie et Immunoanalyse (SPI), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), and Deeptope SAS, Massy, France
Subjects: [SDV.BIO]Life Sciences [q-bio]/Biotechnology, yeast surface display, SARS-CoV-2, [SDV]Life Sciences [q-bio], Immunology, Antibody engineering, COVID-19, Single-Domain Antibodies, Antibodies, Viral, Antibodies, Neutralizing, nanobodies, deep mutational scanning, Spike Glycoprotein, Coronavirus, Immunology and Allergy, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Antigenic Drift and Shift, ComputingMilieux_MISCELLANEOUS, Protein Binding
Abstract: International audience; Here, we report the molecular engineering of nanobodies that bind with picomolar affinity to both SARS-CoV-1 and SARS-CoV-2 receptor-binding domains (RBD) and are highly neutralizing. We applied deep mutational engineering to VHH72, a nanobody initially specific for SARS-CoV-1 RBD with little cross-reactivity to SARS-CoV-2 antigen. We first identified all the individual VHH substitutions that increase binding to SARS-CoV-2 RBD and then screened highly focused combinatorial libraries to isolate engineered nanobodies with improved properties. The corresponding VHH-Fc molecules show high affinities for SARS-CoV-2 antigens from various emerging variants and SARS-CoV-1, block the interaction between ACE2 and RBD, and neutralize the virus with high efficiency. Its rare specificity across sarbecovirus relies on its peculiar epitope outside the immunodominant regions. The engineered nanobodies share a common motif of three amino acids, which contribute to the broad specificity of recognition. Our results show that deep mutational engineering is a very powerful method, especially to rapidly adapt existing antibodies to new variants of pathogens.
Published: 2022
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9. Monitoring In Vivo Performances of Protein–Drug Conjugates Using Site-Selective Dual Radiolabeling and Ex Vivo Digital Imaging

Author: Héloïse Cahuzac, Antoine Sallustrau, Carole Malgorn, Fabrice Beau, Peggy Barbe, Victor Babin, Steven Dubois, Alberto Palazzolo, Robert Thai, Isabelle Correia, Ki Baek Lee, Sébastien Garcia-Argote, Olivier Lequin, Mathilde Keck, Hervé Nozach, Sophie Feuillastre, Xin Ge, Gregory Pieters, Davide Audisio, Laurent Devel, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Service de Chimie Bio-Organique et de Marquage (SCBM), Laboratoire des biomolécules (LBM UMR 7203), Chimie Moléculaire de Paris Centre (FR 2769), École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Département de Chimie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and The University of Texas Health Science Center at Houston (UTHealth)
Subjects: Immunoconjugates, [SDV]Life Sciences [q-bio], Drug Discovery, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, Molecular Medicine, Carbon Radioisotopes, Article
Abstract: International audience; In preclinical models, the development and optimization of protein−drug conjugates require accurate determination of the plasma and tissue profiles of both the protein and its conjugated drug. To this aim, we developed a bioanalytical strategy based on dual radiolabeling and ex vivo digital imaging. By combining enzymatic and chemical reactions, we obtained homogeneous dual-labeled anti-MMP-14 Fabs (antigen-binding fragments) conjugated to monomethyl auristatin E where the protein scaffold was labeled with carbon-14 (14C) and the conjugated drug with tritium (3H). These antibody−drug conjugates with either a noncleavable or a cleavable linker were then evaluated in vivo. By combining liquid scintillation counting and ex vivo dual-isotope radio-imaging, it was possible not only to monitor both components simultaneously during their circulation phase but also to quantify accurately their amount accumulated within the different organs.
Published: 2022
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10. Editorial: Pharmacological Aspects of Ligand-Gated Ion Channels as Targets of Natural and Synthetic Agents

Author: César Mattei, Hélène Tricoire-Leignel, Christian Legros, Richard J. Lewis, Jordi Molgó, MitoVasc - Physiopathologie Cardiovasculaire et Mitochondriale (MITOVASC), Université d'Angers (UA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
Subjects: [SDV]Life Sciences [q-bio], General Neuroscience, ComputingMilieux_MISCELLANEOUS
Abstract: International audience
Published: 2022
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11. Practical Synthesis of Phosphinic Dipeptides by Tandem Esterification of Aminophosphinic and Acrylic Acids under Silylating Conditions

Author: Paraskevi Kokkala, Kostas Voreakos, Angelos Lelis, Konstantinos Patiniotis, Nikolaos Skoulikas, Laurent Devel, Angeliki Ziotopoulou, Eleni Kaloumenou, Dimitris Georgiadis, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and University of Athens, Athens
Subjects: phosphinic, pseudopeptides, P-Michael, silylation, hexamethyldisilazane, inhibitors, Zn-metalloproteases, acrylic acids, aminophosphinic, organophosphorus, Acrylates, Esterification, Chemistry (miscellaneous), Organic Chemistry, Drug Discovery, Molecular Medicine, Pharmaceutical Science, [CHIM]Chemical Sciences, Dipeptides, Physical and Theoretical Chemistry, Phosphinic Acids, Analytical Chemistry
Abstract: International audience; In this report, a synthetic protocol for the preparation of phosphinic dipeptides of type 5 is presented. These compounds serve as valuable building blocks for the development of highly potent phosphinopeptidic inhibitors of medicinally relevant Zn-metalloproteases and aspartyl proteases. The proposed method is based on the tandem esterification of α-aminophosphinic and acrylic acids under silylating conditions in order to subsequently participate in a P-Michael reaction. The scope of the transformation was investigated by using a diverse set of readily available acrylic acids and (R)-α-aminophosphinic acids, and high yields were achieved in all cases. In most examples reported herein, the isolation of biologically relevant (R,S)-diastereoisomers became possible by simple crystallization from the crude products, thus enhancing the operational simplicity of the proposed method. Finally, functional groups corresponding to acidic or basic natural amino acids are also compatible with the reaction conditions. Based on the above, we expect that the practicality of the proposed protocol will facilitate the discovery of pharmacologically useful bioactive phosphinic peptides.
Published: 2022
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12. Vasorin Plays a Critical Role in Vascular Smooth Muscle Cells and Arterial Functions

Author: Loïc Louvet, Gaëlle Lenglet, A. Michaela Krautzberger, Romuald Mentaverri, Frédéric Hague, Clara Kowalewski, Nassim Mahtal, Julie Lesieur, Anne‐Laure Bonnet, Caroline Andrique, Céline Gaucher, Cathy Gomila, Heinrich Schrewe, Pierre‐Louis Tharaux, Said Kamel, Catherine Chaussain, Isabelle Six, Marrow Adiposity & Bone Lab - Adiposité Médullaire et Os - ULR 4490 (MABLab (ex-pmoi)), Université du Littoral Côte d'Opale (ULCO)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), Mécanismes physiopathologiques et conséquences des calcifications vasculaires - UR UPJV 7517 (MP3CV), Université de Picardie Jules Verne (UPJV)-CHU Amiens-Picardie, Laboratoire de Physiologie Cellulaire et Moléculaire - UR UPJV 4667 (LPCM), Université de Picardie Jules Verne (UPJV), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Service de Chimie Bio-Organique et de Marquage (SCBM), Pathologies, imagerie et biothérapies oro-faciales = Orofacial pathologies, imaging and biotherapies (URP 2496), Université Paris Cité (UPCité), CHU Charles Foix [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), UFR Odontologie [Santé] - Université Paris Cité (UFR Odontologie UPCité), and Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: Mice, Knockout, Physiology, Angiotensin II, Myocytes, Smooth Muscle, Clinical Biochemistry, Membrane Proteins, Cell Biology, Nitric Oxide, Muscle, Smooth, Vascular, Mice, Phenylephrine, Tamoxifen, Animals, Humans, Calcium, Carbachol, Apoptosis Regulatory Proteins, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract: International audience; Within the cardiovascular system, the protein vasorin (Vasn) is predominantly expressed by vascular smooth muscle cells (VSMCs) in the coronary arteries and the aorta. Vasn knockout (Vasn(-/-) ) mice die within 3 weeks of birth. In the present study, we investigated the role of vascular Vasn expression on vascular function. We used inducible Vasn knockout mice (Vasn(CRE-ERT KO) and Vasn(SMMHC-CRE-ERT2 KO) , in which respectively all cells or SMCs only are targeted) to analyze the consequences of total or selective Vasn loss on vascular function. Furthermore, in vivo effects were investigated in vitro using human VSMCs. The death of Vasn(CRE-ERT KO) mice 21 days after tamoxifen injection was concomitant with decreases in blood pressure, angiotensin II levels, and vessel contractibility to phenylephrine. The Vasn(SMMHC-CRE-ERT2 KO) mice displayed concomitant changes in vessel contractibility in response to phenylephrine and angiotensin II levels. In vitro, VASN deficiency was associated with a shift toward the SMC contractile phenotype, an increase in basal intracellular Ca(2+) levels, and a decrease in the SMCs' ability to generate a calcium signal in response to carbachol or phenylephrine. Additionally, impaired endothelium-dependent relaxation (due to changes in nitric oxide signaling) was observed in all Vasn knockout mice models. Our present findings highlight the role played by Vasn SMC expression in the maintenance of vascular functions. The mechanistic experiments suggested that these effects are mediated by SMC phenotype switching and changes in intracellular calcium homeostasis, angiotensin II levels, and NO signaling.
Published: 2022
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13. Role of metalloproteases in the CD95 signaling pathways

Author: Laurent Devel, Nicolas Guedeney, Sarah Bregant, Animesh Chowdhury, Mickael Jean, Patrick Legembre, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut des Sciences Chimiques de Rennes (ISCR), Université de Rennes (UR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Contrôle de la Réponse Immune B et des Lymphoproliférations (CRIBL), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-OmégaHealth (ΩHealth), Université de Limoges (UNILIM)-Université de Limoges (UNILIM), INCa PLBIO [ANR-17-CE15-0027], ANR PRCE, ITMO Cancer of Aviesan within the framework of the 2021-2030 Cancer Control Strategy, [PLBIO 2018-132], and ANR-17-CE15-0027,FASILE,Impact du CD95L, dans la dysfonction des lymphocytes T exprimant l'IL-17 au cours du lupus(2017)
Subjects: MMP, inflammation, Immunology, Immunology and Allergy, ADAM, CD95L, cancer, [CHIM]Chemical Sciences, cleavage
Abstract: International audience; CD95L (also known as FasL or CD178) is a member of the tumor necrosis family (TNF) superfamily. Although this transmembrane ligand has been mainly considered as a potent apoptotic inducer in CD95 (Fas)-expressing cells, more recent studies pointed out its role in the implementation of non-apoptotic signals. Accordingly, this ligand has been associated with the aggravation of inflammation in different auto-immune disorders and in the metastatic occurrence in different cancers. Although it remains to decipher all key factors involved in the ambivalent role of this ligand, accumulating clues suggest that while the membrane bound CD95L triggers apoptosis, its soluble counterpart generated by metalloprotease-driven cleavage is responsible for its non-apoptotic functions. Nonetheless, the metalloproteases (MMPs and ADAMs) involved in the CD95L shedding, the cleavage sites and the different stoichiometries and functions of the soluble CD95L remain to be elucidated. To better understand how soluble CD95L triggers signaling pathways from apoptosis to inflammation or cell migration, we propose herein to summarize the different metalloproteases that have been described to be able to shed CD95L, their cleavage sites and the biological functions associated with the released ligands. Based on these new findings, the development of CD95/CD95L-targeting therapeutics is also discussed.
Published: 2022
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14. Report from the 27th (Virtual) Meeting on Toxinology, 'Toxins: Mr Hyde or Dr Jekyll?', Organized by the French Society of Toxinology, 9–10 December 2021

Author: Daniel Ladant, Pascale Marchot, Sylvie Diochot, Gilles Prévost, Michel R. Popoff, Evelyne Benoit, Biochimie des Interactions Macromoléculaires / Biochemistry of Macromolecular Interactions, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Architecture et fonction des macromolécules biologiques (AFMB), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut de pharmacologie moléculaire et cellulaire (IPMC), Université Nice Sophia Antipolis (1965 - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Centre National de la Recherche Scientifique (CNRS)-Université Côte d'Azur (UCA), Virulence bactérienne précoce : fonctions cellulaires et contrôle de l'infection aiguë et subaiguë, Université de Strasbourg (UNISTRA), Toxines bactériennes - Bacterial Toxins, Institut Pasteur [Paris] (IP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), SIMOPRO, Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPC), and Institut Pasteur [Paris]-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPC)
Subjects: plant toxin, maine toxin, toxin structure, medical application, Health, Toxicology and Mutagenesis, [SDV.TOX]Life Sciences [q-bio]/Toxicology, bacterial toxin, animal toxin, Toxicology, toxin receptor, toxin function
Abstract: International audience; The French Society of Toxinology (SFET) organized its 27th annual meeting on 9–10 December 2021 as a virtual meeting (e-RT27). The central theme of this meeting was “Toxins: Mr Hyde or Dr Jekyll?”, emphasizing the latest findings on plant, fungal, algal, animal and bacterial toxins during 10 lectures, 15 oral communications (shorter lectures) and 20 posters shared by ca. 80 participants. The abstracts of lectures and posters, as well as the winners of the best oral communication and poster awards, are presented in this report.
Published: 2022
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15. A new Kunitz‐type snake toxin family associated with an original mode of interaction with the vasopressin 2 receptor

Author: Laura Droctové, Justyna Ciolek, Christiane Mendre, Amélia Chorfa, Paola Huerta, Chrystelle Carvalho, Charlotte Gouin, Manon Lancien, Goran Stanajic‐Petrovic, Lorine Braco, Guillaume Blanchet, Gregory Upert, Gregory De Pauw, Peggy Barbe, Mathilde Keck, Gilles Mourier, Bernard Mouillac, Servent Denis, Ricardo C. Rodríguez de la Vega, Loïc Quinton, Nicolas Gilles, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut de Génomique Fonctionnelle (IGF), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Laboratory of Mass Spectrometry, GIGA, Université de Liège, Ecologie Systématique et Evolution (ESE), AgroParisTech-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), This work was supported by the French Atomic and Alternative Energies and La Ligue contre le Cancer for financial support and for the Amelia Chorfa PhD programme funding., and Keck, Mathilde
Subjects: Pharmacology, Proteomics, Receptors, Vasopressin, Peptide sequencing, Snake venom, Vasopressins, [SDV]Life Sciences [q-bio], Antagonist, V2R, Kunitz peptide, Rats, [SDV] Life Sciences [q-bio], Peptide synthesis, [SDV.SP.PHARMA] Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, [SDV.TOX]Life Sciences [q-bio]/Toxicology, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, Animals, Elapidae, Peptides, ComputingMilieux_MISCELLANEOUS, Snake Venoms
Abstract: International audience; Background and Purpose: Venomous animals express numerous Kunitz-type peptides. The mambaquaretin-1 (MQ1) peptide identified from the Dendroaspis angusticeps venom is the most selective antagonist of the arginine-vasopressin V2 receptor (V2R) and the only unique Kunitz-type peptide active on a GPCR. We aimed to exploit other mamba venoms to enlarge the V2R-Kunitz peptide family and gain insight into the MQ1 molecular mode of action.Experimental Approach: We used a bio-guided screening assay to identify novel MQs and placed them phylogenetically. MQs were produced by solid-phase peptide synthesis and characterized in vitro by binding and functional tests and in vivo by diuresis measurement in rats.Key Results: Eight additional MQs were identified with nanomolar affinities for the V2R, all antagonists. MQs form a new subgroup in the Kunitz family, close to the V2R non-active dendrotoxins and to two V2R-active cobra toxins. Sequence comparison between active and non-active V2R Kunitz peptides highlighted five positions, among which four are involved in V2R interaction and belong to the two large MQ1 loops. We finally determined that eight positions, part of these two loops, interact with the V2R. The variant MQ1-K39A showed a higher affinity for the hV2R, but not for the rat V2R.Conclusions and Implications: A new function and mode of action is associated with the Kunitz peptides. The number of MQ1 residues involved in V2R binding is large and may explain its absolute selectivity. MQ1-K39A represents the first step in the improvement of the MQ1 design from a medicinal perspective.
Published: 2022
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16. C910 chemical compound inhibits the traffiking of several bacterial AB toxins with cross-protection against influenza virus

Author: Yu Wu, Nassim Mahtal, Eléa Paillares, Léa Swistak, Sara Sagadiev, Mridu Acharya, Caroline Demeret, Sylvie Van Der Werf, Florence Guivel-Benhassine, Olivier Schwartz, Serena Petracchini, Amel Mettouchi, Lucie Caramelle, Pierre Couvineau, Robert Thai, Peggy Barbe, Mathilde Keck, Priscille Brodin, Arnaud Machelart, Valentin Sencio, François Trottein, Martin Sachse, Gaëtan Chicanne, Bernard Payrastre, Florian Ville, Victor Kreis, Michel-Robert Popoff, Ludger Johannes, Jean-Christophe Cintrat, Julien Barbier, Daniel Gillet, Emmanuel Lemichez, TROTTEIN, François, Caractérisation d'un composé antitoxines bactériennes à large spectre - - ToxProtect_Larg_Spectr2019 - ANR-19-ASTR-0001 - ASTRID - VALID, Integrative Biology of Emerging Infectious Diseases - - IBEID2010 - ANR-10-LABX-0062 - LABX - VALID, Research Laboratory on Drugs and Therapeutic Innovation - - LERMIT2010 - ANR-10-LABX-0033 - LABX - VALID, Toxines bactériennes - Bacterial Toxins, Université Paris Cité (UPCité)-Microbiologie Intégrative et Moléculaire (UMR6047), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Service de Chimie Bio-Organique et de Marquage (SCBM), Seattle Children's Research Institute [Seattle, WA, USA], Génétique Moléculaire des Virus à ARN - Molecular Genetics of RNA Viruses (GMV-ARN (UMR_3569 / U-Pasteur_2)), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Virus et Immunité - Virus and immunity, Université Paris-Saclay, Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 (CIIL), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS), CHU Lille, Plateforme BioImagerie Ultrastructurale – Ultrastructural BioImaging Platform (UTechS UBI), Institut Pasteur [Paris] (IP), Institut des Maladies Métaboliques et Casdiovasculaires (UPS/Inserm U1297 - I2MC), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM), Chimie biologique des membranes et ciblage thérapeutique (CBMCT - UMR 3666 / U1143), Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), This work was funded by the joint ministerial program of Research and Development against Chemical, Biological, Radiological, Nuclear and Explosive risks ( CBRNE ), grants from the Agence Nationale de la Recherche ( ANR-19-ASTR-0001 ), Labex IBEID ( ANR-10-LABX-62-IBEID ), and the French Alternative Energies and Atomic Energy Commission ( CEA ). N.M., E.P., L.C., and F.V. were supported by DGA-MRIS/AID scholarships with CEA or Pasteur. SIMoS and SCBM are members of the Laboratory of Excellence LERMIT and were supported by a grant from the Agence Nationale de la Recherche ( ANR-10-LABX-33 )., ANR-19-ASTR-0001,ToxProtect_Larg_Spectr,Caractérisation d'un composé antitoxines bactériennes à large spectre(2019), ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010), ANR-10-LABX-0033,LERMIT,Research Laboratory on Drugs and Therapeutic Innovation(2010), Virus et Immunité - Virus and immunity (CNRS-UMR3569), and Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: [SDV] Life Sciences [q-bio], Biological sciences, Multidisciplinary, [SDV]Life Sciences [q-bio], Microbiology
Abstract: International audience; The development of anti-infectives against a large range of AB-like toxin-producing bacteria includes the identification of compounds disrupting toxin transport through both the endolysosomal and retrograde pathways. Here, we performed a high-throughput screening of compounds blocking Rac1 proteasomal degradation triggered by the Cytotoxic Necrotizing Factor-1 (CNF1) toxin, which was followed by orthogonal screens against two toxins that hijack the endolysosomal (diphtheria toxin) or retrograde (Shiga-like toxin 1) pathways to intoxicate cells. This led to the identification of the molecule C910 that induces the enlargement of EEA1-positive early endosomes associated with sorting defects of CNF1 and Shiga toxins to their trafficking pathways. C910 protects cells against eight bacterial AB toxins and the CNF1-mediated pathogenic Escherichia coli invasion. Interestingly, C910 reduces influenza A H1N1 and SARS-CoV-2 viral infection in vitro. Moreover, parenteral administration of C910 to mice resulted in its accumulation in lung tissues and a reduction in lethal influenza infection.
Published: 2022
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17. Toxins: Mr Hyde or Dr Jekyll?

Author: Daniel Ladant, Gilles Prévost, Popoff, Michel R., Evelyne BENOIT, Biochimie des Interactions Macromoléculaires / Biochemistry of Macromolecular Interactions, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Virulence bactérienne précoce : fonctions cellulaires et contrôle de l'infection aiguë et subaiguë, Université de Strasbourg (UNISTRA), Centre National de Référence des Bactéries Anaérobies et Botulisme - National Reference Center Anaerobic Bacteria and Botulism (CNR), Institut Pasteur [Paris], Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), ERL CNRS 9004, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), D. LADANT, G. PREVOST, M.R. POPOFF & E. BENOIT (Eds), and Toxins
Subjects: Bacterial toxins, [SDV]Life Sciences [q-bio], [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Medical application, Animal toxins, Plant toxins, Toxin receptor/target, Toxinogenesis regulation, Toxin structure and organization, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Marine toxins, Toxin function/activity, Phylogeny, Toxin delivery into cell
Abstract: International audience; Toxins are biologically active substances produced by all kinds of living organisms such as bacteria, fungi, plants, and animals. They present a vast diversity of molecular structures and target a wide variety of receptors linked to physiological processes. Selected during evolution to give them a disabling/lethal effect, they display finely tuned functional properties in terms of affinity and selectivity. Toxins are, therefore, very attractive compounds because they exhibit a Janus-like character, being both poisons and remedies, and, as a matter of fact, have been primarily investigated not only for the light they can throw on fundamental physiological processes, but also for their potential therapeutic applications. The Toxins Special Issue entitled “Toxins: Mr Hyde or Dr Jekyll?” will accept manuscripts describing recent advances concerning toxins of various origins (such as animal, bacterial, fungi, marine and plant toxins) and a variety of topics covering toxin structure and organization, toxin receptor/target, phylogeny, toxinogenesis regulation, medical application, toxin function/activity, and toxin delivery into cell. This Special Issue is open (but not limited) to oral communications and posters that will be presented during the virtual meeting of the French Society of Toxinology (SFET) that will be held on 9th and 10th December 2021 (e-RT27—27th SFET meeting).
Published: 2022

18. Report from the 28th Meeting on Toxinology, 'Toxins: What’s up, Doc?', Organized by the French Society of Toxinology on 28–29 November 2022

Author: Pascale Marchot, Évelyne Benoit, Ziad Fajloun, Sylvie Diochot, Architecture et fonction des macromolécules biologiques (AFMB), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), SIMOPRO, Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Azm Center for research in biotechnology and its applications [TRIPOLI], Université Libanaise, Institut de pharmacologie moléculaire et cellulaire (IPMC), Université Nice Sophia Antipolis (1965 - 2019) (UNS), and COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Centre National de la Recherche Scientifique (CNRS)-Université Côte d'Azur (UCA)
Subjects: Toxinomics, Health, Toxicology and Mutagenesis, Medical application, Neuromuscular junction, Toxin structure–function relationship, Deep learning, Bacterial toxin, Toxin receptor–target, Toxicology, Toxin tracking, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Marine toxin, Plant toxin, Toxin structure, Animal toxin
Abstract: International audience; The French Society of Toxinology (SFET) organized its 28th annual meeting on 28–29 November 2022 (RT28). The central theme of this meeting was “Toxins: What’s up, Doc?”, emphasizing the latest findings on animal, bacterial, algal, plant and fungal toxins through sessions dedicated to deep learning, toxin tracking and toxinomic advances, shared by ca. 80 participants. The abstracts of the 10 invited and 11 selected lectures and 15 posters, along with the names of the Best Oral Communication and Best Poster awardees, are presented in this report.
Published: 2023
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19. Synthetic Analogues of Huwentoxin-IV Spider Peptide With Altered Human NaV1.7/NaV1.6 Selectivity Ratios

Author: Ludivine Lopez, Jérôme Montnach, Barbara Oliveira-Mendes, Kuldip Khakh, Baptiste Thomas, Sophia Lin, Cécile Caumes, Steven Wesolowski, Sébastien Nicolas, Denis Servent, Charles Cohen, Rémy Béroud, Evelyne Benoit, Michel De Waard, unité de recherche de l'institut du thorax UMR1087 UMR6291 (ITX), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN), Xenon Pharmaceuticals, Smartox Biotechnology, Université Paris-Saclay, CEA, SIMoS, Gif-sur-Yvette, France, ERL CNRS 9004, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Unité de recherche de l'institut du thorax (ITX-lab), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), SIMOPRO, Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), ANR-11-LABX-0015,ICST,Canaux ioniques d'intérêt thérapeutique(2011), and ANR-18-CE19-0024,OptChemCom,Technologies intégrées d' optique, photochimie et informatique pour étudier la synergie physiologique des canaux ioniques(2018)
Subjects: 0303 health sciences, huwentoxin-IV analogues, NaV1.6 and NaV1.7 channel subtypes, QH301-705.5, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, [SDV]Life Sciences [q-bio], Cyriopagopus schmidti, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, Cell Biology, development of pain therapeutics, structure-function relationship, 03 medical and health sciences, Cell and Developmental Biology, 0302 clinical medicine, peptide synthesis, Biology (General), automated patch-clamp, 030217 neurology & neurosurgery, 030304 developmental biology, Developmental Biology, Original Research
Abstract: Huwentoxin-IV (HwTx-IV), a peptide discovered in the venom of the Chinese bird spider Cyriopagopus schmidti, has been reported to be a potent antinociceptive compound due to its action on the genetically-validated NaV1.7 pain target. Using this peptide for antinociceptive applications in vivo suffers from one major drawback, namely its negative impact on the neuromuscular system. Although studied only recently, this effect appears to be due to an interaction between the peptide and the NaV1.6 channel subtype located at the presynaptic level. The aim of this work was to investigate how HwTx-IV could be modified in order to alter the original human (h) NaV1.7/NaV1.6 selectivity ratio of 23. Nineteen HwTx-IV analogues were chemically synthesized and tested for their blocking effects on the Na+ currents flowing through these two channel subtypes stably expressed in cell lines. Dose-response curves for these analogues were generated, thanks to the use of an automated patch-clamp system. Several key amino acid positions were targeted owing to the information provided by earlier structure-activity relationship (SAR) studies. Among the analogues tested, the potency of HwTx-IV E4K was significantly improved for hNaV1.6, leading to a decreased hNaV1.7/hNaV1.6 selectivity ratio (close to 1). Similar decreased selectivity ratios, but with increased potency for both subtypes, were observed for HwTx-IV analogues that combine a substitution at position 4 with a modification of amino acid 1 or 26 (HwTx-IV E1G/E4G and HwTx-IV E4K/R26Q). In contrast, increased selectivity ratios (>46) were obtained if the E4K mutation was combined to an additional double substitution (R26A/Y33W) or simply by further substituting the C-terminal amidation of the peptide by a carboxylated motif, linked to a marked loss of potency on hNaV1.6 in this latter case. These results demonstrate that it is possible to significantly modulate the selectivity ratio for these two channel subtypes in order to improve the potency of a given analogue for hNaV1.6 and/or hNaV1.7 subtypes. In addition, selective analogues for hNaV1.7, possessing better safety profiles, were produced to limit neuromuscular impairments.
Published: 2021
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20. Structural and Functional Diversity of Animal Toxins Interacting With GPCRs

Author: Anne-Cécile Van Baelen, Philippe Robin, Pascal Kessler, Arhamatoulaye Maïga, Nicolas Gilles, Denis Servent, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
Subjects: GPCR, QH301-705.5, [SDV]Life Sciences [q-bio], toxins, peptides, venom, structure, Biology (General), Biochemistry, Genetics and Molecular Biology (miscellaneous), Molecular Biology, Biochemistry
Abstract: International audience; Peptide toxins from venoms have undergone a long evolutionary process allowing host defense or prey capture and making them highly selective and potent for their target. This has resulted in the emergence of a large panel of toxins from a wide diversity of species, with varied structures and multiple associated biological functions. In this way, animal toxins constitute an inexhaustible reservoir of druggable molecules due to their interesting pharmacological properties. One of the most interesting classes of therapeutic targets is the G-protein coupled receptors (GPCRs). GPCRs represent the largest family of membrane receptors in mammals with approximately 800 different members. They are involved in almost all biological functions and are the target of almost 30% of drugs currently on the market. Given the interest of GPCRs in the therapeutic field, the study of toxins that can interact with and modulate their activity with the purpose of drug development is of particular importance. The present review focuses on toxins targeting GPCRs, including peptide-interacting receptors or aminergic receptors, with a particular focus on structural aspects and, when relevant, on potential medical applications. The toxins described here exhibit a great diversity in size, from 10 to 80 amino acids long, in disulfide bridges, from none to five, and belong to a large panel of structural scaffolds. Particular toxin structures developed here include inhibitory cystine knot (ICK), three-finger fold, and Kunitz-type toxins. We summarize current knowledge on the structural and functional diversity of toxins interacting with GPCRs, concerning first the agonist-mimicking toxins that act as endogenous agonists targeting the corresponding receptor, and second the toxins that differ structurally from natural agonists and which display agonist, antagonist, or allosteric properties.
Published: 2021
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21. Gambierol blocks a K$^+$ current fraction and affects action potential duration and the firing rate in rat fetal chromaffin cells

Author: Molgó, Jordi, Bournaud, Roland, Schlumberger, Sébastien, Sasaki, Makoto, Fuwa, Haruhiko, Benoit, Evelyne, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), SIMOPRO, Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Laboratoire de neurobiologie cellulaire et moléculaire (NBCM), Centre National de la Recherche Scientifique (CNRS), Department of Biomolecular Sciences, Graduate School of Life Sciences, Tohoku University [Sendai], Chuo University (Chuo University), This study was supported in part by the project ALERTOX-NET (EAPA_317/2016), funded by the Interreg Atlantic program, and in part by the CNRS., The International Society for the Study of Harmful Algae (ISSHA), Band Schmidt C.J., and Rodríguez Gómez C.F.
Subjects: Gambierol, marine biotoxin, [SDV.TOX]Life Sciences [q-bio]/Toxicology, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], potassium channels
Abstract: The dinoflagellate genus Gambierdiscus, particularly Gambierdiscus toxicus is known to produce numerous ladder polycyclic ether compounds, including gambierol, characterized by a transfused octacyclic polyether core, as well as the identified ciguatoxins responsible for ciguatera poisoning. Gambierol inhibits voltage-gated K+ (KV) channels in various mammalian excitable and nonexcitable cells, and motor nerve terminals of the neuromuscular junction. In the present study, we investigated the action of gambierol on outward K+ currents of cultured rat fetal adrenal medulla chromaffin cells using the whole-cell configuration voltage-clamp method. The pharmacological dissection of the outward K+ current, using selective K+ channel inhibitors, revealed that gambierol reduced just a small fraction of the total outward current without affecting calcium-activated K+ currents that were sensitive to apamin and iberiotoxin, and ATP-sensitive K+ currents sensitive to glibenclamide. Cultured fetal chromaffin cells were excitable and generated either evoked (upon direct electric stimulation) or spontaneous action potentials. These action potentials were sensitive to tetrodotoxin that blocks voltage-gated Na+ (NaV) channels. Gambierol neither affected the rising phase nor the overshoot of action potentials but significantly prolonged their repolarizing phase and increased the firing rate of action potentials during sustained current depolarization, as determined using current-clamp recordings. Our results highlight the effects of gambierol on KV channels and on the electrical properties of rat fetal chromaffin cells. It is likely that gambierol may cross the placental barrier as ciguatoxins do. Further studies are needed to determine whether such actions may have deleterious effects on fetuses.
Published: 2021
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22. Medica-Plus: a Micromegas-based proof-of-concept detector for sub-becquerel tritium activity assessment at the service of oncological research

Author: F. Jambon, S. Aune, P. Baron, T. Benoit, T. Bey, D. Desforge, E. Ferrer-Ribas, A. Grabas, M. Kebbiri, I. Mandjavidze, T. Papaevangelou, M. Riallot, M. Vandenbroucke, F. Beau, V. Dive, C. Malgorn, F. Malloggi, A. Rousselot, F. Carrel, M. Trocmé, Institut de Recherches sur les lois Fondamentales de l'Univers (IRFU), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Laboratoire Interdisciplinaire sur l'Organisation Nanométrique et Supramoléculaire (LIONS), Nanosciences et Innovation pour les Matériaux, la Biomédecine et l'Energie (ex SIS2M) (NIMBE UMR 3685), Institut Rayonnement Matière de Saclay (IRAMIS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut Rayonnement Matière de Saclay (IRAMIS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Intégration des Systèmes et des Technologies (LIST (CEA)), Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), and Laboratoire d'Intégration des Systèmes et des Technologies (LIST)
Subjects: 0303 health sciences, Nuclear and High Energy Physics, Physics - Instrumentation and Detectors, FOS: Physical sciences, Instrumentation and Detectors (physics.ins-det), 010501 environmental sciences, 01 natural sciences, 3. Good health, 03 medical and health sciences, H radiolabelling, [PHYS.PHYS.PHYS-INS-DET]Physics [physics]/Physics [physics]/Instrumentation and Detectors [physics.ins-det], Instrumentation, Micromegas, 030304 developmental biology, 0105 earth and related environmental sciences, MPGD
Abstract: To fulfill needs in oncological research a new Micromegas detector has been developed to follow radiolabelled drugs in living organisms at the single cell level. This article describes the proof-of-concept of such a detector and compares its ability to detect and assess sub-becquerel \tritium~activities with a commercial $\beta$-imager, Comment: 14 pages, to be submitted to NIMA
Published: 2021
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23. Guidance Level for Brevetoxins in French Shellfish

Author: Arnich, Nathalie, Abadie, Eric, Amzil, Zouher, Dechraoui Bottein, Marie-Yasmine, Comte, Katia, Chaix, Estelle, Delcourt, Nicolas, Hort, Vincent, Mattei, César, Molgó, Jordi, Le Garrec, Raphaele, Direction de l'Evaluation des Risques (DER), Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), MARine Biodiversity Exploitation and Conservation (UMR MARBEC), Institut de Recherche pour le Développement (IRD)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER), Ecology and Conservation Science for Sustainable Seas (ECOSEAS), Centre National de la Recherche Scientifique (CNRS)-Université Côte d'Azur (UCA), Molécules de Communication et Adaptation des Micro-organismes (MCAM), Muséum national d'Histoire naturelle (MNHN)-Centre National de la Recherche Scientifique (CNRS), Centre antipoison et de toxicovigilance [CHU Toulouse] (CAPTV Toulouse), Pôle Médecine d'urgences [CHU Toulouse], Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), Laboratoire de sécurité des aliments de Maisons-Alfort (LSAl), MitoVasc - Physiopathologie Cardiovasculaire et Mitochondriale (MITOVASC), Université d'Angers (UA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Laboratoire sur les interactions Epithéliums Neurones (LIEN), Université de Brest (UBO), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Institut de Recherche pour le Développement (IRD), Centre antipoison et de toxicovigilance (Toulouse) (CAPTV Toulouse), CHU Toulouse [Toulouse]-CHU Purpan, Physiopathologie Cardiovasculaire et Mitochondriale (MITOVASC), and GUITTON, SOPHIE
Subjects: guidance level, QH301-705.5, Oxocins, food and beverages, [SDV.TOX.TCA]Life Sciences [q-bio]/Toxicology/Toxicology and food chain, Article, [SDV.AEN] Life Sciences [q-bio]/Food and Nutrition, brevetoxins, shellfish, [SDV.TOX.TCA] Life Sciences [q-bio]/Toxicology/Toxicology and food chain, Dinoflagellida, Mediterranean Sea, emerging toxins, Animals, Humans, Shellfish Poisoning, marine biotoxins, Marine Toxins, France, Biology (General), neurotoxic shellfish poisoning, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, Environmental Monitoring
Abstract: To prevent health risks associated with the consumption of shellfish contaminated with BTXs in France, a working group was set up by the French Agency for Food, Environmental and Occupational Health & Safety (Anses). One of the aims of this working group was to propose a guidance level for the presence of BTXs in shellfish.; International audience; Brevetoxins (BTXs) are marine biotoxins responsible for neurotoxic shellfish poisoning (NSP) after ingestion of contaminated shellfish. NSP is characterized by neurological, gastrointestinal and/or cardiovascular symptoms. The main known producer of BTXs is the dinoflagellate Karenia brevis, but other microalgae are also suspected to synthesize BTX-like compounds. BTXs are currently not regulated in France and in Europe. In November 2018, they have been detected for the first time in France in mussels from a lagoon in the Corsica Island (Mediterranean Sea), as part of the network for monitoring the emergence of marine biotoxins in shellfish. To prevent health risks associated with the consumption of shellfish contaminated with BTXs in France, a working group was set up by the French Agency for Food, Environmental and Occupational Health & Safety (Anses). One of the aims of this working group was to propose a guidance level for the presence of BTXs in shellfish. Toxicological data were too limited to derive an acute oral reference dose (ARfD). Based on human case reports, we identified two lowest-observed-adverse-effect levels (LOAELs). A guidance level of 180 µg BTX-3 eq./kg shellfish meat is proposed, considering a protective default portion size of 400 g shellfish meat.
Published: 2021
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24. Electrophysiological Evaluation of Pacific Oyster (Crassostrea gigas) Sensitivity to Saxitoxin and Tetrodotoxin

Author: Boullot, Floriane, Fabioux, Caroline, Hégaret, Hélène, Boudry, Pierre, Soudant, Philippe, Benoit, Evelyne, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Laboratoire des Sciences de l'Environnement Marin (LEMAR) (LEMAR), Institut de Recherche pour le Développement (IRD)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de Brest (UBO)-Institut Universitaire Européen de la Mer (IUEM), Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER), SIMOPRO, Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), This research was funded by the French National Research Agency (ANR) [13-CESA-0019-ACCUTOX], the French Research Network PHYCOTOX, the French National Center for Scientific Research (CNRS), and the French Alternative Energies and Atomic Energy Commission (CEA). F.B. was supported by a pre-doctoral fellowship from the Région Bretagne and the Université de Bretagne Occidentale (UBO)., ANR-13-CESA-0019,ACCUTOX,De la caractérisation des déterminants de l'accumulation des toxines paralysantes (PST) chez l'huître (Crassostrea gigas) au risque sanitaire pour l'homme dans son contexte sociétal(2013), Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut Français de Recherche pour l'Exploitation de la Mer - Brest (IFREMER Centre de Bretagne), ERL CNRS 9004, and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)
Subjects: animal structures, QH301-705.5, Alexandrium minutum, [SDV]Life Sciences [q-bio], fungi, food and beverages, Crassostrea gigas, equipment and supplies, paralytic shellfish toxins, [SDV.TOX]Life Sciences [q-bio]/Toxicology, compound nerve action potential, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Biology (General), geographic locations
Abstract: Pacific oysters (Crassostrea gigas) may bio-accumulate high levels of paralytic shellfish toxins (PST) during harmful algal blooms of the genus Alexandrium. These blooms regularly occur in coastal waters, affecting oyster health and marketability. The aim of our study was to analyse the PST-sensitivity of nerves of Pacific oysters in relation with toxin bio-accumulation. The results show that C. gigas nerves have micromolar range of saxitoxin (STX) sensitivity, thus providing intermediate STX sensitivity compared to other bivalve species. However, theses nerves were much less sensitive to tetrodotoxin. The STX-sensitivity of compound nerve action potential (CNAP) recorded from oysters experimentally fed with Alexandrium minutum (toxic-alga-exposed oysters), or Tisochrysis lutea, a non-toxic microalga (control oysters), revealed that oysters could be separated into STX-resistant and STX-sensitive categories, regardless of the diet. Moreover, the percentage of toxin-sensitive nerves was lower, and the STX concentration necessary to inhibit 50% of CNAP higher, in recently toxic-alga-exposed oysters than in control bivalves. However, no obvious correlation was observed between nerve sensitivity to STX and the STX content in oyster digestive glands. None of the nerves isolated from wild and farmed oysters was detected to be sensitive to tetrodotoxin. In conclusion, this study highlights the good potential of cerebrovisceral nerves of Pacific oysters for electrophysiological and pharmacological studies. In addition, this study shows, for the first time, that C. gigas nerves have micromolar range of STX sensitivity. The STX sensitivity decreases, at least temporary, upon recent oyster exposure to dinoflagellates producing PST under natural, but not experimental environment.
Published: 2021
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25. Reprogramming Escherichia coli for the production of prenylated indole diketopiperazine alkaloids

Author: Nicolas Canu, Muriel Gondry, Isabelle Correia, Pascal Belin, Robert Thai, Olivier Lequin, Isabelle Jacques, Pavlina Dubois, Steven G. DuBois, Fabien Le Chevalier, Mireille Moutiez, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Laboratoire des biomolécules (LBM UMR 7203), Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Département de Chimie - ENS Paris, École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Chimie Moléculaire de Paris Centre (FR 2769), Institut de Chimie du CNRS (INC)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP), This work was supported by the French Alternative Energies and Atomic Energy Commission (CEA), the French National Centre for Scientific Research (CNRS), the Sorbonne University and the French National Research Agency (grant ANR-16-CE29-0026)., We thank Dr Taek Soon Lee and Melanya Gudzeva from the Lawrence Berkeley National Laboratory for providing us with plasmids pJBEI-3085 and pJBEI-3122., ANR-16-CE29-0026,DKP-COMBIBIO,L'utilisation des voies de biosynthèse dépendant des synthases de cyclodipeptides pour obtenir de nouvelles 2,5-dicétopipérazines bioactives(2016), Chimie Moléculaire de Paris Centre (FR 2769), École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Département de Chimie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université Pierre et Marie Curie - Paris 6 (UPMC), Institute of Neurosciences and Psychology [Glasgow], University of Glasgow, Institut de Neurosciences de la Timone (INT), and Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)
Subjects: 0301 basic medicine, Stereochemistry, [SDV]Life Sciences [q-bio], lcsh:Medicine, Context (language use), Diketopiperazines, medicine.disease_cause, Article, Indole Alkaloids, 03 medical and health sciences, 0302 clinical medicine, medicine, Escherichia coli, Moiety, [CHIM]Chemical Sciences, Author Correction, lcsh:Science, 2. Zero hunger, chemistry.chemical_classification, Indole test, Prenylation, Multidisciplinary, Chemistry, Alkaloid, lcsh:R, Bioproduction, 030104 developmental biology, Enzyme, Biocatalysis, lcsh:Q, Mevalonate pathway, Genetic Engineering, Metabolic engineering, 030217 neurology & neurosurgery
Abstract: Prenylated indole diketopiperazine (DKP) alkaloids are important bioactive molecules or their precursors. In the context of synthetic biology, efficient means for their biological production would increase their chemical diversification and the discovery of novel bioactive compounds. Here, we prove the suitability of the Escherichia coli chassis for the production of prenylated indole DKP alkaloids. We used enzyme combinations not found in nature by co-expressing bacterial cyclodipeptide synthases (CDPSs) that assemble the DKP ring and fungal prenyltransferases (PTs) that transfer the allylic moiety from the dimethylallyl diphosphate (DMAPP) to the indole ring of tryptophanyl-containing cyclodipeptides. Of the 11 tested combinations, seven resulted in the production of eight different prenylated indole DKP alkaloids as determined by LC-MS/MS and NMR characterization. Two were previously undescribed. Engineering E. coli by introducing a hybrid mevalonate pathway for increasing intracellular DMAPP levels improved prenylated indole DKP alkaloid production. Purified product yields of 2–26 mg/L per culture were obtained from culture supernatants. Our study paves the way for the bioproduction of novel prenylated indole DKP alkaloids in a tractable chassis that can exploit the cyclodipeptide diversity achievable with CDPSs and the numerous described PT activities.
Published: 2019
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26. Di-phosphorylated BAF shows altered structural dynamics and binding to DNA, but interacts with its nuclear envelope partners

Author: Virginie Ropars, Guillaume Hoffmann, Sophie Zinn-Justin, Marie-Hélène Le Du, Philippe Cuniasse, Ambre Petitalot, Robert Thai, Camille Samson, Agathe Marcelot, Stevens Dubois, José A. Márquez, Simona Miron, François-Xavier Theillet, Département Plateforme (PF I2BC), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), High Throughput Crystallization Laboratory, European Molecular Biology Laboratory Grenoble Outstation, European Molecular Biology Laboratory, ANR-10-INBS-0005,FRISBI,Infrastructure Française pour la Biologie Structurale Intégrée(2010), European Project: 653706,H2020,H2020-INFRAIA-2014-2015,iNEXT(2015), European Project: 871037,iNEXT-Discovery (H2020-EU.1.4.1.2), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), and European Molecular Biology Laboratory [Grenoble] (EMBL)
Subjects: AcademicSubjects/SCI00010, Emerin, Protein Serine-Threonine Kinases, Biology, Protein Structure, Secondary, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, [CHIM.ANAL]Chemical Sciences/Analytical chemistry, Genetics, Humans, Amino Acid Sequence, Phosphorylation, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Ternary complex, Gene, 030304 developmental biology, 0303 health sciences, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Kinase, Gene regulation, Chromatin and Epigenetics, Intracellular Signaling Peptides and Proteins, Membrane Proteins, Nuclear Proteins, DNA, Cell cycle, Lamin Type A, 3. Good health, Cell biology, DNA-Binding Proteins, chemistry, ddc:540, Protein Processing, Post-Translational, 030217 neurology & neurosurgery, Lamin, Protein Binding
Abstract: Barrier-to-autointegration factor (BAF), encoded by the BANF1 gene, is an abundant and ubiquitously expressed metazoan protein that has multiple functions during the cell cycle. Through its ability to cross-bridge two double-stranded DNA (dsDNA), it favours chromosome compaction, participates in post-mitotic nuclear envelope reassembly and is essential for the repair of large nuclear ruptures. BAF forms a ternary complex with the nuclear envelope proteins lamin A/C and emerin, and its interaction with lamin A/C is defective in patients with recessive accelerated aging syndromes. Phosphorylation of BAF by the vaccinia-related kinase 1 (VRK1) is a key regulator of BAF localization and function. Here, we demonstrate that VRK1 successively phosphorylates BAF on Ser4 and Thr3. The crystal structures of BAF before and after phosphorylation are extremely similar. However, in solution, the extensive flexibility of the N-terminal helix α1 and loop α1α2 in BAF is strongly reduced in di-phosphorylated BAF, due to interactions between the phosphorylated residues and the positively charged C-terminal helix α6. These regions are involved in DNA and lamin A/C binding. Consistently, phosphorylation causes a 5000-fold loss of affinity for dsDNA. However, it does not impair binding to lamin A/C Igfold domain and emerin nucleoplasmic region, which leaves open the question of the regulation of these interactions.
Published: 2021
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27. An antibody targeting type III secretion system induces broad protection against Salmonella and Shigella infections

Author: Raphaël Sierocki, Bernard Maillere, Maria Lucia Orsini Delgado, Stéphanie Simon, Hervé Nozach, Bakhos Jneid, Marc Plaisance, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Service de Pharmacologie et Immunoanalyse (SPI), and PhD program of the Commissariat a l'Energie Atomique et aux Energies Alternatives
Subjects: 0301 basic medicine, Serotype, Bacterial Diseases, Salmonella typhimurium, Salmonella, Salmonellosis, Physiology, RC955-962, medicine.disease_cause, Pathology and Laboratory Medicine, Biochemistry, Type three secretion system, Shigella flexneri, Mice, Medical Conditions, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Arctic medicine. Tropical medicine, Immune Physiology, Medicine and Health Sciences, Type III Secretion Systems, Shigella, Enzyme-Linked Immunoassays, Mice, Inbred BALB C, Immune System Proteins, biology, Bacterial Gastroenteritis, Eukaryota, Antibodies, Monoclonal, Antibodies, Bacterial, 3. Good health, Gastroenteritis, Bacterial Pathogens, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Infectious Diseases, Shigellosis, Medical Microbiology, Female, Public aspects of medicine, RA1-1270, Pathogens, Research Article, Neglected Tropical Diseases, Immunology, Virulence, Gastroenterology and Hepatology, Research and Analysis Methods, Microbiology, Antibodies, 03 medical and health sciences, Enterobacteriaceae, medicine, Animals, Humans, Immunoassays, Microbial Pathogens, Dysentery, Bacillary, Antigens, Bacterial, Salmonella Infections, Animal, 030102 biochemistry & molecular biology, Bacteria, Public Health, Environmental and Occupational Health, Organisms, Fungi, Biology and Life Sciences, Proteins, biology.organism_classification, medicine.disease, Tropical Diseases, Yeast, 030104 developmental biology, Immunologic Techniques, Conformational epitope, HeLa Cells
Abstract: Salmonella and Shigella bacteria are food- and waterborne pathogens that are responsible for enteric infections in humans and are still the major cause of morbidity and mortality in the emerging countries. The existence of multiple Salmonella and Shigella serotypes as well as the emergence of strains resistant to antibiotics requires the development of broadly protective therapies. Recently, the needle tip proteins of the type III secretion system of these bacteria were successfully utilized (SipD for Salmonella and IpaD for Shigella) as vaccine immunogens to provide good prophylactic cross-protection in murine models of infections. From these experiments, we have isolated a cross-protective monoclonal antibody directed against a conserved region of both proteins. Its conformational epitope determined by Deep Mutational Scanning is conserved among needle tip proteins of all pathogenic Shigella species and Salmonella serovars, and are well recognized by this antibody. Our study provides the first in vivo experimental evidence of the importance of this common region in the mechanism of virulence of Salmonella and Shigella and opens the way to the development of cross-protective therapeutic agents., Author summary Salmonella and Shigella are responsible for gastrointestinal diseases and continue to remain a serious health hazard in South and South-East Asia and African countries, even more with the new emergence of multi drug resistances. Developed vaccines are either not commercialized (for Shigella) or cover only a limited number of serotypes (for Salmonella). There is thus a crucial need to develop cross-protective therapies. By targeting proteins SipD and IpaD belonging respectively to the injectisome of Salmonella and Shigella and necessary to their virulence, we have shown that a monoclonal antibody (mAb) directed against a conserved common region of their apical part provides good cross-protection prophylactic efficacy. We have determined the region targeted by this mAb which could explain why it is conserved among Salmonella and Shigella bacteria.
Published: 2021
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28. Inhibition of InsP3R with Xestospongin B Reduces Mitochondrial Respiration and Induces Selective Cell Death in T Cell Acute Lymphoblastic Leukemia Cells

Author: Daniela Sauma, Alenka Lovy, Galdo Bustos, Ulises Ahumada-Castro, César Cárdenas, Jordi Molgó, Pablo Cruz, Universidad Mayor [Santiago de Chile], Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Universidad de Chile = University of Chile [Santiago] (UCHILE), University of California [Santa Barbara] (UC Santa Barbara), University of California (UC), and This research was funded by FONDECYT #1200255, 1180385, CONICYT/FONDAP # 15150012 and CONICYT Doctoral Fellowship Program funds PC (#21180306)
Subjects: MESH: Cell Death, 0301 basic medicine, Mitochondrion, Precursor T-Cell Lymphoblastic Leukemia-Lymphoma, bioenergetics, Jurkat cells, lcsh:Chemistry, chemistry.chemical_compound, MESH: Macrocyclic Compounds, 0302 clinical medicine, Inositol 1,4,5-Trisphosphate Receptors, Inositol, Receptor, Oxazoles, lcsh:QH301-705.5, Spectroscopy, Cell Death, MESH: Oxazoles, [SDV.MHEP.HEM]Life Sciences [q-bio]/Human health and pathology/Hematology, General Medicine, Computer Science Applications, Mitochondria, MESH: Precursor T-Cell Lymphoblastic Leukemia-Lymphoma, MESH: Leukocytes, Mononuclear, medicine.anatomical_structure, 030220 oncology & carcinogenesis, T-ALL, Programmed cell death, MESH: Cell Line, Tumor, Macrocyclic Compounds, MESH: Mitochondria, T cell, Cell Respiration, [SDV.CAN]Life Sciences [q-bio]/Cancer, Catalysis, Article, Inorganic Chemistry, 03 medical and health sciences, MESH: Inositol 1,4,5-Trisphosphate Receptors, Cell Line, Tumor, medicine, Humans, cancer, Physical and Theoretical Chemistry, Molecular Biology, MESH: Humans, calcium, Endoplasmic reticulum, Organic Chemistry, 030104 developmental biology, chemistry, lcsh:Biology (General), lcsh:QD1-999, Cell culture, MESH: Biomarkers, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, Cancer research, Leukocytes, Mononuclear, MESH: Cell Respiration, metabolism, Biomarkers
Abstract: International audience; T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematological malignancy whose chemoresistance and relapse persist as a problem despite significant advances in its chemotherapeutic treatments. Mitochondrial metabolism has emerged as an interesting therapeutic target given its essential role in maintaining bioenergetic and metabolic homeostasis. T-ALL cells are characterized by high levels of mitochondrial respiration, making them suitable for this type of intervention. Mitochondrial function is sustained by a constitutive transfer of calcium from the endoplasmic reticulum to mitochondria through the inositol 1,4,5-trisphosphate receptor (InsP3R), making T-ALL cells vulnerable to its inhibition. Here, we determine the bioenergetic profile of the T-ALL cell lines CCRF-CEM and Jurkat and evaluate their sensitivity to InsP3R inhibition with the specific inhibitor, Xestospongin B (XeB). Our results show that T-ALL cell lines exhibit higher mitochondrial respiration than non-malignant cells, which is blunted by the inhibition of the InsP3R. Prolonged treatment with XeB causes T-ALL cell death without affecting the normal counterpart. Moreover, the combination of XeB and glucocorticoids significantly enhanced cell death in the CCRF-CEM cells. The inhibition of InsP3R with XeB rises as a potential therapeutic alternative for the treatment of T-ALL.
Published: 2021
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29. Gambierol Blocks a K+ Current Fraction without Affecting Catecholamine Release in Rat Fetal Adrenomedullary Cultured Chromaffin Cells

Author: Evelyne Benoit, Sébastien Schlumberger, Jordi Molgó, Makoto Sasaki, Haruhiko Fuwa, Roland Bournaud, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut des Sciences du Vivant Frédéric JOLIOT (JOLIOT), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Laboratoire de neurobiologie cellulaire et moléculaire (NBCM), Centre National de la Recherche Scientifique (CNRS), Tohoku University [Sendai], and Chuo University (Chuo University)
Subjects: fetal adrenomedullary chromaffin cell, gambierol, potassium currents, calcium-activated K+ channels, ATP-sensitive K+ channels, catecholamine release, ATP-sensitive K$^+$ channels, [SDV.TOX]Life Sciences [q-bio]/Toxicology, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Health, Toxicology and Mutagenesis, Toxicology
Abstract: International audience; Gambierol inhibits voltage-gated K+ (KV) channels in various excitable and non-excitable cells. The purpose of this work was to study the effects of gambierol on single rat fetal (F19–F20) adrenomedullary cultured chromaffin cells. These excitable cells have different types of KV channels and release catecholamines. Perforated whole-cell voltage-clamp recordings revealed that gambierol (100 nM) blocked only a fraction of the total outward K$^+$ current and slowed the kinetics of K$^+$ current activation. The use of selective channel blockers disclosed that gambierol did not affect calcium-activated K$^+$ (KCa) and ATP-sensitive K$^+$ (KATP) channels. The gambierol concentration necessary to inhibit 50% of the K$^+$ current-component sensitive to the polyether (IC50) was 5.8 nM. Simultaneous whole-cell current-clamp and single-cell amperometry recordings revealed that gambierol did not modify the membrane potential following 11s depolarizing current-steps, in both quiescent and active cells displaying repetitive firing of action potentials, and it did not increase the number of exocytotic catecholamine release events, with respect to controls. The subsequent addition of apamin and iberiotoxin, which selectively block the KCa channels, both depolarized the membrane and enhanced by 2.7 and 3.5-fold the exocytotic event frequency in quiescent and active cells, respectively. These results highlight the important modulatory role played by KCa channels in the control of exocytosis from fetal (F19–F20) adrenomedullary chromaffin cells.
Published: 2022
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30. Backbone Cyclization Turns a Venom Peptide into a Stable and Equipotent Ligand at Both Muscle and Neuronal Nicotinic Receptors

Author: Norelle L. Daly, Muriel Amar, Adèle Faucherre, Julien Giribaldi, Jordi Molgó, Casey Schmidt, Denis Servent, David Wilson, Hamid Moha Ou Maati, Sébastien Dutertre, Annette Nicke, Edward R. J. Evans, Anna Durner, Yves Haufe, Christine Enjalbal, Institut des Biomolécules Max Mousseron [Pôle Chimie Balard] (IBMM), Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Institut de Chimie du CNRS (INC)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Ludwig Maximilian University [Munich] (LMU), James Cook University (JCU), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut de Génomique Fonctionnelle (IGF), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Service de Pharmacologie et Immunoanalyse (SPI), ANR-16-CE34-0002,SPIDERBEE,Découverte et caractérisation de ligands peptidiques pour l'étude la neurobiologie de l'abeille Apis mellifera et pour la conception d'insecticides écologiques(2016), Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS), and DMTS, Université Paris Saclay, CEA, INRAE, SPI
Subjects: [SDV]Life Sciences [q-bio], Peptide, Venom, Nicotinic Antagonists, Pharmacology, Receptors, Nicotinic, Ligands, 01 natural sciences, 03 medical and health sciences, Mice, Drug Discovery, Potency, Animals, Amino Acid Sequence, IC50, Zebrafish, 030304 developmental biology, chemistry.chemical_classification, Neurons, 0303 health sciences, biology, Venoms, Muscles, biology.organism_classification, Ligand (biochemistry), 0104 chemical sciences, Bioavailability, Protein Structure, Tertiary, 010404 medicinal & biomolecular chemistry, Nicotinic acetylcholine receptor, nervous system, chemistry, Cyclization, Larva, Molecular Medicine, Conotoxins, Peptides, Locomotion, Muscle Contraction, Protein Binding
Abstract: International audience; Venom peptides are promising drug leads, but their therapeutic use is often limited by stability and bioavailability issues. In this study, we designed cyclic analogues of α-conotoxin CIA, a potent muscle nicotinic acetylcholine receptor (nAChR) blocker with a significantly lower affinity at the neuronal α3β2 subtype. Remarkably, all analogues retained the low nanomolar activity of native CIA toward muscle-type nAChRs but showed greatly improved resistance to degradation in human serum and, surprisingly, displayed up to 52-fold higher potency for the α3β2 neuronal nAChR subtype (IC50 1.3 nM). Comparison of nuclear magnetic resonance-derived structures revealed some differences that might explain the gain of potency at α3β2 nAChRs. All peptides were highly paralytic when injected into adult zebrafish and bath-applied to zebrafish larvae, suggesting barrier-crossing capabilities and efficient uptake. Finally, these cyclic CIA analogues were shown to be unique pharmacological tools to investigate the contribution of the presynaptic α3β2 nAChR subtype to the train-of-four fade.
Published: 2020
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31. Alterations of peripheral nerve excitability in an experimental autoimmune encephalomyelitis mouse model for multiple sclerosis

Author: Teixeira, Nathalia Bernardes, Picolo, Gisele, Giardini, Aline Carolina, Boumezbeur, Fawzi, Pottier, Géraldine, Kuhnast, Bertrand, Servent, Denis, Benoit, Evelyne, Instituto Butantan [São Paulo], Service NEUROSPIN (NEUROSPIN), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), LaBoratoire d'Imagerie biOmédicale MultimodAle Paris-Saclay (BIOMAPS), Service Hospitalier Frédéric Joliot (SHFJ), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), SIMOPRO, Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Unité BioMaps (BIOMAPS), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Experimental autoimmune encephalomyelitis, Encephalomyelitis, Autoimmune, Experimental, Mouse, Research, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Neural Conduction, Neuromuscular junction, Action Potentials, lcsh:RC346-429, Peptide Fragments, Electrophysiology, Multiple sclerosis, Mice, Myelin oligodendrocyte glycoprotein, Disease Progression, Animals, Myelin-Oligodendrocyte Glycoprotein, Peripheral Nerves, Peripheral nervous system, Muscle, Skeletal, lcsh:Neurology. Diseases of the nervous system, Myelin Sheath
Abstract: Background Experimental autoimmune encephalomyelitis (EAE) is the most commonly used and clinically relevant murine model for human multiple sclerosis (MS), a demyelinating autoimmune disease characterized by mononuclear cell infiltration into the central nervous system (CNS). The aim of the present study was to appraise the alterations, poorly documented in the literature, which may occur at the peripheral nervous system (PNS) level. Methods To this purpose, a multiple evaluation of peripheral nerve excitability was undertaken, by means of a minimally invasive electrophysiological method, in EAE mice immunized with the myelin oligodendrocyte glycoprotein (MOG) 35-55 peptide, an experimental model for MS that reproduces, in animals, the anatomical and behavioral alterations observed in humans with MS, including CNS inflammation, demyelination of neurons, and motor abnormalities. Additionally, the myelin sheath thickness of mouse sciatic nerves was evaluated using transmission electronic microscopy. Results As expected, the mean clinical score of mice, daily determined to describe the symptoms associated to the EAE progression, increased within about 18 days after immunization for EAE mice while it remained null for all control animals. The multiple evaluation of peripheral nerve excitability, performed in vivo 2 and 4 weeks after immunization, reveals that the main modifications of EAE mice, compared to control animals, are a decrease of the maximal compound action potential (CAP) amplitude and of the stimulation intensity necessary to generate a CAP with a 50% maximum amplitude. In addition, and in contrast to control mice, at least 2 CAPs were recorded following a single stimulation in EAE animals, reflecting various populations of sensory and motor nerve fibers having different CAP conduction speeds, as expected if a demyelinating process occurred in the PNS of these animals. In contrast, single CAPs were always recorded from the sensory and motor nerve fibers of control mice having more homogeneous CAP conduction speeds. Finally, the myelin sheath thickness of sciatic nerves of EAE mice was decreased 4 weeks after immunization when compared to control animals. Conclusions In conclusion, the loss of immunological self-tolerance to MOG in EAE mice or in MS patients may not be only attributed to the restricted expression of this antigen in the immunologically privileged environment of the CNS but also of the PNS.
Published: 2020
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32. Monoaryl derivatives as transthyretin fibril formation inhibitors: Design, synthesis, biological evaluation and structural analysis

Author: Elisabetta Orlandini, Elisa Nuti, William Shepard, Armando Rossello, Lidia Ciccone, Nicolo Tonali, Susanna Nencetti, Caterina Camodeca, Carole Fruchart-Gaillard, University of Pisa - Università di Pisa, Synchrotron SOLEIL (SSOLEIL), Centre National de la Recherche Scientifique (CNRS), Biomolécules : Conception, Isolement, Synthèse (BioCIS), Institut de Chimie du CNRS (INC)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-CY Cergy Paris Université (CY), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Ministero dell’Istruzione, dell’Universitá e della Ricerca (PRIN 2017SNRXH3)
Subjects: Models, Molecular, endocrine system, Clinical Biochemistry, Molecular Conformation, Pharmaceutical Science, Diflunisal, Fibril formation inhibitors, Monoaryl derivatives, Transthyretin, X-ray TTR-ligand complexes, Crystallography, X-Ray, 01 natural sciences, Biochemistry, Protein Aggregates, Structure-Activity Relationship, Tetramer, Alzheimer Disease, Drug Discovery, Extracellular, medicine, Humans, Prealbumin, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Binding site, Molecular Biology, Binding Sites, biology, 010405 organic chemistry, Chemistry, Organic Chemistry, nutritional and metabolic diseases, Fibrillogenesis, Small molecule, In vitro, 3. Good health, 0104 chemical sciences, 010404 medicinal & biomolecular chemistry, Neuroprotective Agents, biology.protein, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, Molecular Medicine, Propionates, medicine.drug, Protein Binding
Abstract: International audience; Transthyretin (TTR) is a beta-sheet-rich homotetrameric protein that transports thyroxine (T4) and retinol both in plasma and in cerebrospinal fluid. TTR also interacts with amyloid-beta, playing a protective role in Alzheimer's disease. Dissociation of the native transthyretin (TTR) tetramer is widely accepted as the critical step in TTR amyloids fibrillogenesis, and is responsible for extracellular deposition of amyloid fibrils. Small molecules, able to bind in T4 binding sites and stabilize the TTR tetramer, are interesting tools to treat and prevent systemic ATTR amyloidosis. We report here the synthesis, in vitro evaluation and three-dimensional crystallographic analyses of new monoaryl-derivatives in complex with TTR. Of the derivatives reported here, the best inhibitor of TTR fibrillogenesis, 1d, exhibits an activity similar to diflunisal.
Published: 2020
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33. Structure–Activity Relationship Studies of Retro-1 Analogues against Shiga Toxin

Author: Lilia Ait Ouarab, Valérie Pons, David Montoir, Julien Barbier, Flora Ngadjeua, David-Alexandre Buisson, Jean-Christophe Cintrat, Alexandra Clerget, Daniel Gillet, Aurélien Michau, Lucie Caramelle, Hajer Abdelkafi, Service de Chimie Bio-Organique et de Marquage (SCBM), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), joint ministerial program of R&D against CBRNe risks, and ANR-11-BSV2-0018,RETROscreen,Génétique chimique de la voie du transport rétrograde(2011)
Subjects: endocrine system, Endosome, Golgi Apparatus, Shiga Toxins, 01 natural sciences, 03 medical and health sciences, chemistry.chemical_compound, symbols.namesake, Structure-Activity Relationship, Drug Discovery, Structure–activity relationship, Humans, Cytotoxicity, 030304 developmental biology, EC50, 0303 health sciences, Benzodiazepinones, biology, Shiga toxin, Golgi apparatus, 0104 chemical sciences, 010404 medicinal & biomolecular chemistry, Protein Transport, Ricin, chemistry, Biochemistry, biology.protein, symbols, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, Molecular Medicine, Intracellular, HeLa Cells
Abstract: International audience; High-throughput screening has shown that Retro-1 inhibits ricin and Shiga toxins by diminishing their intracellular trafficking via the retrograde route, from early endosomes to the Golgi apparatus. To improve the activity of Retro-1, a structure-activity relationship (SAR) study was undertaken and yielded an analogue with a roughly 70-fold better half-maximal effective concentration (EC50) against Shiga toxin cytotoxicity measured in a cell protein synthesis assay.
Published: 2020
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34. Cancer cells with defective oxidative phosphorylation require endoplasmic reticulum–to–mitochondria Ca(2+) transfer for survival

Author: Pablo Cruz, Eduardo Silva-Pavez, Melany Rios, Paola Murgas, Fabian Jaňa, Elizabeth Mendoza, Oscar Cerda, Alenka Lovy, Irene Georgakoudi, Galdo Bustos, J. Kevin Foskett, Armen Zakarian, Paula Farias, Martin Hunter, Hernan Huerta, Félix A. Urra, Ulises Ahumada-Castro, Craig Mizzoni, Jordi Molgó, César Cárdenas, Instituto Antartico Chileno, Universidad Mayor [Santiago de Chile], CONICET/Universidad Nacional de Jujuy, Tufts University School of Medicine [Boston], Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
Subjects: Programmed cell death, Cell Survival, [SDV]Life Sciences [q-bio], education, Dehydrogenase, Oxidative phosphorylation, Mitochondrion, Endoplasmic Reticulum, Biochemistry, Article, Oxidative Phosphorylation, Mitochondrial Proteins, 03 medical and health sciences, 0302 clinical medicine, Cell Line, Tumor, Neoplasms, Humans, Molecular Biology, health care economics and organizations, 030304 developmental biology, 0303 health sciences, Chemistry, Endoplasmic reticulum, Cell Biology, Cell biology, Mitochondria, Neoplasm Proteins, 030220 oncology & carcinogenesis, Cancer cell, Calcium, NAD+ kinase, Flux (metabolism)
Abstract: Spontaneous Ca(2+) signaling from the InsP(3)R intracellular Ca(2+) release channel to mitochondria is essential for optimal oxidative phosphorylation (OXPHOS) and ATP production. In cells with defective OXPHOS, reductive carboxylation replaces oxidative metabolism to maintain amounts of reducing equivalents and metabolic precursors. To investigate the role of mitochondrial Ca(2+) uptake in regulating bioenergetics in these cells, we used OXPHOS-competent and OXPHOS-defective cells. Inhibition of InsP(3)R activity or mitochondrial Ca(2+) uptake increased α-ketoglutarate (αKG) abundance and the NAD(+)/NADH ratio, indicating that constitutive endoplasmic reticulum (ER)–to–mitochondria Ca(2+) transfer promoted optimal αKG dehydrogenase (αKGDH) activity. Reducing mitochondrial Ca(2+) inhibited αKGDH activity and increased NAD(+), which induced SIRT1-dependent autophagy in both OXPHOS-competent and OXPHOS-defective cells. Whereas autophagic flux in OXPHOS-competent cells promoted cell survival, it was impaired in OXPHOS-defective cells because of inhibition of autophagosome-lysosome fusion. Inhibition of αKGDH and impaired autophagic flux in OXPHOS-defective cells resulted in pronounced cell death in response to interruption of constitutive flux of Ca(2+) from ER to mitochondria. These results demonstrate that mitochondria play a fundamental role in maintaining bioenergetic homeostasis of both OXPHOS-competent and OXPHOS-defective cells, with Ca(2+) regulation of αKGDH activity playing a pivotal role. Inhibition of ER-to-mitochondria Ca(2+) transfer may represent a general therapeutic strategy against cancer cells regardless of their OXPHOS status.
Published: 2020
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35. Specificity of the T Cell Response to Protein Biopharmaceuticals

Author: Moustafa Hamze, Aurélien Azam, Marie de Bourayne, Catherine Menier, Evelyne Correia, Sylvain Meunier, Bernard Maillere, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Innovative Medicines Initiative Joint Undertaking ABIRISK (Anti-Biopharmaceutical Immunization Risk) project under grant agreement #115303, European Union’s Seventh Framework Program (FP7/2007-2013), Labex in Research on Medication and Therapeutic Innovation (LERMIT), and CEA
Subjects: 0301 basic medicine, lcsh:Immunologic diseases. Allergy, Isoantigens, T cell, T-Lymphocytes, Cell, Immunology, Epitopes, T-Lymphocyte, T cell selection, T-Cell Antigen Receptor Specificity, Review, Thymus Gland, Biology, immunogenicity, Epitope, Antibodies, 03 medical and health sciences, 0302 clinical medicine, Immune system, T-Lymphocyte Subsets, medicine, Immunology and Allergy, therapeutic protein, Animals, Humans, Clonal Selection, Antigen-Mediated, epitope, Biological Products, tolerance, Factor VIII, Effector, Immunogenicity, Antibodies, Monoclonal, Proteins, biopharmaceuticals, 3. Good health, 030104 developmental biology, medicine.anatomical_structure, therapeutic antibody, biology.protein, [SDV.IMM]Life Sciences [q-bio]/Immunology, Cytokines, Antibody, lcsh:RC581-607, 030215 immunology
Abstract: International audience; The anti-drug antibody (ADA) response is an undesired humoral response raised against protein biopharmaceuticals (BPs) which can dramatically disturb their therapeutic properties. One particularity of the ADA response resides in the nature of the immunogens, which are usually human(ized) proteins and are therefore expected to be tolerated. CD4 T cells initiate, maintain and regulate the ADA response and are therefore key players of this immune response. Over the last decade, advances have been made in characterizing the T cell responses developed by patients treated with BPs. Epitope specificity and phenotypes of BP-specific T cells have been reported and highlight the effector and regulatory roles of T cells in the ADA response. BP-specific T cell responses are assessed in healthy subjects to anticipate the immunogenicity of BP prior to their testing in clinical trials. Immunogenicity prediction, also called preclinical immunogenicity assessment, aims at identifying immunogenic BPs and immunogenic BP sequences before any BP injection in humans. All of the approaches that have been developed to date rely on the detection of BP-specific T cells in donors who have never been exposed to BPs. The number of BP-specific T cells circulating in the blood of these donors is therefore limited. T cell assays using cells collected from healthy donors might reveal the weak tolerance induced by BPs, whose endogenous form is expressed at a low level. These BPs have a complete human sequence, but the level of their endogenous form appears insufficient to promote the negative selection of autoreactive T cell clones. Multiple T cell epitopes have also been identified in therapeutic antibodies and some other BPs. The pattern of identified T cell epitopes differs across the antibodies, notwithstanding their humanized, human or chimeric nature. However, in all antibodies, the non-germline amino acid sequences mainly found in the CDRs appear to be the main driver of immunogenicity, provided they can be presented by HLA class II molecules. Considering the fact that the BP field is expanding to include new formats and gene and cell therapies, we face new challenges in understanding and mastering the immunogenicity of new biological products.
Published: 2020
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36. Health risk assessment related to pinnatoxins in French shellfish

Author: Jean-Paul Vernoux, Marie-Bénédicte Peyrat, Thomas Maignien, Eric Abadie, Vincent Hort, Jordi Molgó, Nathalie Arnich, Jean-Marc Fremy, Valérie Fessard, Emmeline Lagrange, Nicolas Delcourt, César Mattei, Direction de l'Evaluation des Risques (DER), Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), MARine Biodiversity Exploitation and Conservation (UMR MARBEC), Institut de Recherche pour le Développement (IRD)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Toulouse Neuro Imaging Center (ToNIC), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Toulouse Mind & Brain Institut (TMBI), Université Toulouse - Jean Jaurès (UT2J), Université de Toulouse (UT)-Université de Toulouse (UT)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université Toulouse - Jean Jaurès (UT2J), Université de Toulouse (UT)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT), Laboratoire de Fougères - ANSES, Retraité, Laboratoire de sécurité des aliments de Maisons-Alfort (LSAl), Centre Hospitalier Universitaire [Grenoble] (CHU), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université de Caen Normandie (UNICAEN), Normandie Université (NU), MitoVasc - Physiopathologie Cardiovasculaire et Mitochondriale (MITOVASC), Université d'Angers (UA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Institut de Recherche pour le Développement (IRD), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Hôpital Purpan [Toulouse], CHU Toulouse [Toulouse]-CHU Toulouse [Toulouse], Physiopathologie Cardiovasculaire et Mitochondriale (MITOVASC), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Toulouse Mind & Brain Institut (TMBI), Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées, and CCSD, Accord Elsevier
Subjects: 0106 biological sciences, biotoxine marine, Emerging marine biotoxins, Zoology, Pinnatoxins, 010501 environmental sciences, Biology, Toxicology, 01 natural sciences, pinnatoxine, Dietary Exposure, Mice, Animals, Humans, Shellfish Poisoning, 14. Life underwater, Adverse effect, Shellfish, Risk assessment, 0105 earth and related environmental sciences, Health risk assessment, business.industry, 010604 marine biology & hydrobiology, marine biotoxin, Dinoflagellate, risk assessment, sécurté des aliments, Food safety, biology.organism_classification, Monitoring program, Acute toxicity, Bivalvia, [SDV.TOX] Life Sciences [q-bio]/Toxicology, shellfish, food safety, coquillage, Seafood, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Toxicity, Dinoflagellida, Marine Toxins, pinnatoxin, France, analyse des risques, business, Environmental Monitoring
Abstract: International audience; Pinnatoxins (PnTXs) are a group of emerging marine biotoxins produced by the benthic dinoflagellate Vulcanodinium rugosum, currently not regulated in Europe or in any other country in the world. In France, PnTXs were detected for the first time in 2011, in mussels from the Ingril lagoon (South of France, Mediterranean coast). Since then, analyses carried out in mussels from this lagoon have shown high concentrations of PnTXs for several months each year. PnTXs have also been detected, to a lesser extent, in mussels from other Mediterranean lagoons and on the Atlantic and Corsican coasts. In the French data, the main analog is PnTX G (low levels of PnTX A are also present in some samples). No cases of PnTXs poisoning in humans have been reported so far in France or anywhere else in the world. In mice, PnTXs induce acute neurotoxic effects, within a few minutes after oral administration. Clinical signs of toxicity include decreased mobility, paralysis of the hind legs, tremors, jumps and breathing difficulties leading to death by respiratory arrest at high doses. The French agency for food safety (ANSES) recently conducted a review of the state of knowledge related to PnTXs and V. rugosum. Based on (i) the clinical signs of toxicity in mice, (ii) the mode of action of PnTXs as nicotinic acetylcholine receptor competitive antagonists and (iii) knowledge on drugs and natural toxins with PnTX-related pharmacology, potential human symptoms have been extrapolated and proposed. In this work, a provisional acute benchmark value for PnTX G of 0.13 μg/kg bw per day has been derived from an oral acute toxicity study in mice. Based on this value and a large shellfish meat portion size of 400g, a concentration lower than 23 μg PnTX G/kg shellfish meat is not expected to result in adverse effects in humans. ANSES recommends taking into account PnTXs in the French official monitoring program for shellfish production and identified data gaps to refine health risk assessment.
Published: 2020
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37. Modulatory effects of pheromone on olfactory learning and memory in moths

Author: Matthieu Dacher, Michel Renou, Valentin Bureau, Nina Deisig, Jeremy Hanoune, Meena Murmu, Abraham Choi, Institut d'écologie et des sciences de l'environnement de Paris (iEES Paris ), Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), University of Cologne, and Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
Subjects: 0106 biological sciences, 0301 basic medicine, Male, Punishment (psychology), Physiology, Olfaction, Biology, Moths, Affect (psychology), 01 natural sciences, 03 medical and health sciences, pheromone, Punishment, Memory, medicine, Animals, Learning, Olfactory memory, Sex Attractants, Sensitization, Appetitive Behavior, olfactory conditioning, proboscis extension response (PER), [SDV.NEU.PC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Psychology and behavior, fungi, [SDV.BA.ZI]Life Sciences [q-bio]/Animal biology/Invertebrate Zoology, Smell, 010602 entomology, 030104 developmental biology, medicine.anatomical_structure, Insect Science, Sex pheromone, plasticity, Pheromone, Olfactory Learning, learning and memory, Neuroscience, olfaction
Abstract: International audience; Pheromones are chemical communication signals known to elicit stereotyped behaviours and/or physiological processes in individuals of the same species, generally in relation to a specific function (e.g. mate finding in moths). However, recent research suggests that pheromones can modulate behaviours, which are not directly related to their usual function and thus potentially affect behavioural plasticity. To test this hypothesis, we studied the possible modulatory effects of pheromones on olfactory learning and memory in Agrotis ipsilon moths, which are well-established models to study sex-pheromones. To achieve this, sexually mature male moths were trained to associate an odour with either a reward (appetitive learning) or punishment (aversive learning) and olfactory memory was tested at medium- and long-term (1h or 1.5h, and 24h). Our results show that male moths can learn to associate an odour with a sucrose reward, as well as a mild electric shock, and that olfactory memory persists over medium- and long-term range. Pheromones facilitated both appetitive and aversive olfactory learning: exposure to the conspecific sex-pheromone before conditioning enhanced appetitive but not aversive learning, while exposure to a sex-pheromone component of a heterospecific species (repellent) facilitated aversive but not appetitive learning. However, this effect was short-term, as medium- and long-term memory were not improved. Thus, in moths, pheromones can modulate olfactory learning and memory, indicating that they contribute to behavioural plasticity allowing optimization of the animal’s behaviour under natural conditions. This might occur through an alteration of sensitization.
Published: 2020
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38. Shiga Toxin-Bearing Microvesicles Exert a Cytotoxic Effect on Recipient Cells Only When the Cells Express the Toxin Receptor

Author: Johansson, Karl, Willysson, Annie, Kristoffersson, Ann-Charlotte, Tontanahal, Ashmita, Gillet, Daniel, Ståhl, Anne-lie, Karpman, Diana, Department of Clinical Sciences [Lund], Lund University [Lund], Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Swedish Research Council K2013-64X-14008-13-5, K2015-99X-22877-01-6 (DK and DG) and 2017-01920, Knut and Alice Wallenberg Foundation (Wallenberg Clinical Scholar 2015.0320), Torsten Söderberg Foundation, Skåne Center of Excellence in Health, IngaBritt and Arne Lundberg’s Research Foundation, Olle Engkvist Byggmästare Foundation, Crown Princess Lovisa’s Society for Child Care, Region Skåne and the Konung Gustaf V:s 80-årsfond (all to DK), and oint ministerial program of R&D against CBRNE risks
Subjects: lcsh:QR1-502, Gb3, Shiga toxin, Shiga Toxin 2, lcsh:Microbiology, retrograde transport, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Cellular and Infection Microbiology, enterohemorrhagic Escherichia coli, Cricetulus, Cricetinae, Hemolytic-Uremic Syndrome, hemolytic uremic syndrome, [SDV.IMM]Life Sciences [q-bio]/Immunology, Animals, Humans, microvesicles, Original Research, globotriaosylceramide, HeLa Cells
Abstract: International audience; Shiga toxin is the main virulence factor of non-invasive enterohemorrhagic Escherichia coli strains capable of causing hemolytic uremic syndrome. Our group has previously shown that the toxin can reach the kidney within microvesicles where it is taken up by renal cells and the vesicles release their cargo intracellularly, leading to toxin-mediated inhibition of protein synthesis and cell death. The aim of this study was to examine if recipient cells must express the globotriaosylceramide (Gb3) toxin receptor for this to occur, or if Gb3-negative cells are also susceptible after uptake of Gb3-positive and toxin-positive microvesicles. To this end we generated Gb3-positive A4GALT-transfected CHO cells, and a vector control lacking Gb3 (CHO-control cells), and decreased Gb3 synthesis in native HeLa cells by exposing them to the glycosylceramide synthase inhibitor PPMP. We used these cells, and human intestinal DLD-1 cells lacking Gb3, and exposed them to Shiga toxin 2-bearing Gb3-positive microvesicles derived from human blood cells. Results showed that only recipient cells that possessed endogenous Gb3 (CHO-Gb3 transfected and native HeLa cells) exhibited cellular injury, reduced cell metabolism and protein synthesis, after uptake of toxin-positive microvesicles. In Gb3positive cells the toxin introduced via vesicles followed the retrograde pathway and was inhibited by the retrograde transport blocker Retro-2.1. CHO-control cells, HeLa cells treated with PPMP and DLD-1 cells remained unaffected by toxin-positive microvesicles. We conclude that Shiga toxin-containing microvesicles can be taken up by Gb3-negative cells but the recipient cell must express endogenous Gb3 for the cell to be susceptible to the toxin.
Published: 2020
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39. Functional dissection of the retrograde Shiga toxin trafficking inhibitor Retro-2

Author: Raphaël Sierocki, Julien Barbier, Adam D. Linstedt, Ludger Johannes, Alison Forrester, Stefan J. Rathjen, Mathilde Munier, Sylvain Pichard, Daniel Gillet, Henri-François Renard, Christophe Lamaze, Damarys Loew, Maria Daniela Garcia-Castillo, Jennifer Martinez, Livia Tepshi, Jean-Christophe Cintrat, Collin Bachert, Audrey Couhert, Florent Dingli, Cesar Augusto Valades-Cruz, Chimie biologique des membranes et ciblage thérapeutique (CBMCT - UMR 3666 / U1143), Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), ANR-19-CE13-0001,GlycoTopoSwitch,La topologie des glycans comme interrupteur moléculaire pour l'activité des intérgines(2019), Carnegie Mellon University [Pittsburgh] (CMU), Service de Chimie Bio-Organique et de Marquage (SCBM), Centre de recherche de l'Institut Curie [Paris], Institut Curie [Paris], Human Frontier Science Program RGP0029-2014, Swedish Research Council European Commission K2015-99X-22877-01-6, Joint Ministerial Program of R&D against CBRNE Risks, Ile de France Région DIM Malinf initiative 140101, Région Ile-de-France, Fondation pour la Recherche Médicale, French Atomic Energy Commission, ANR-11-BSV2-0018,RETROscreen,Génétique chimique de la voie du transport rétrograde(2011), ANR-14-CE16-0004,AntiHUS,Preuve de concept in vivo de molécules contre le syndrome hémolytique et urémique(2014), European Project: 340485,EC:FP7:ERC,ERC-2013-ADG,GALECTCOMPART(2014), Centre National de la Recherche Scientifique (CNRS)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC), Serva, Camille, BLANC - Génétique chimique de la voie du transport rétrograde - - RETROscreen2011 - ANR-11-BSV2-0018 - BLANC - VALID, Appel à projets générique - Preuve de concept in vivo de molécules contre le syndrome hémolytique et urémique - - AntiHUS2014 - ANR-14-CE16-0004 - Appel à projets générique - VALID, La topologie des glycans comme interrupteur moléculaire pour l'activité des intérgines - - GlycoTopoSwitch2019 - ANR-19-CE13-0001 - AAPG2019 - VALID, and Endocytic Membrane Compartmentalization by Galectins - GALECTCOMPART - - EC:FP7:ERC2014-04-01 - 2019-03-31 - 340485 - VALID
Subjects: Retrograde transport, [SDV]Life Sciences [q-bio], Vesicular Transport Proteins, Golgi Apparatus, Endoplasmic Reticulum, Shiga Toxins, chemistry.chemical_compound, anterograde trafficking, COPII, mass spectrometry, 0303 health sciences, biology, Qa-SNARE Proteins, Chemistry, trans-Golgi network, 030302 biochemistry & molecular biology, Shiga-like toxin, Shiga toxin, Transport protein, Cell biology, Protein Transport, chemical genetics, Ricin, SNARE, Benzamides, click chemistry, symbols, Retro-2, GPP130, biological phenomena, cell phenomena, and immunity, retention using selective hooks, syntaxin-5, endocrine system, Sec16A, Endosome, small molecule, chemical biology, Endosomes, Thiophenes, STxB, 03 medical and health sciences, symbols.namesake, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, proximity ligation assay, Molecular Biology, 030304 developmental biology, Endoplasmic reticulum, Biological Transport, Cell Biology, Golgi apparatus, nervous system, Chaperone (protein), Axoplasmic transport, biology.protein, HeLa Cells
Abstract: International audience; The retrograde transport inhibitor Retro-2 has a protective effect on cells and in mice against Shiga-like toxins and ricin. Retro-2 causes toxin accumulation in early endosomes and relocalization of the Golgi SNARE protein syntaxin-5 to the endoplasmic reticulum. The molecular mechanisms by which this is achieved remain unknown. Here, we show that Retro-2 targets the endoplasmic reticulum exit site component Sec16A, affecting anterograde transport of syntaxin-5 from the endoplasmic reticulum to the Golgi. The formation of canonical SNARE complexes involving syntaxin-5 is not affected in Retro-2-treated cells. By contrast, the interaction of syntaxin-5 with a newly discovered binding partner, the retrograde trafficking chaperone GPP130, is abolished, and we show that GPP130 must indeed bind to syntaxin-5 to drive Shiga toxin transport from the endosomes to the Golgi. We therefore identify Sec16A as a druggable target and provide evidence for a non-SNARE function for syntaxin-5 in interaction with GPP130.
Published: 2020
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40. Report from the 26th Meeting on Toxinology, 'Bioengineering of Toxins', Organized by the French Society of Toxinology (SFET) and Held in Paris, France, 4–5 December 2019

Author: Marchot, Pascale, Diochot, Sylvie, Popoff, Michel, Benoit, Evelyne, Architecture et fonction des macromolécules biologiques (AFMB), Institut National de la Recherche Agronomique (INRA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Institut de pharmacologie moléculaire et cellulaire (IPMC), Université Nice Sophia Antipolis (... - 2019) (UNS), Université Côte d'Azur (UCA)-Université Côte d'Azur (UCA)-Centre National de la Recherche Scientifique (CNRS), Toxines bactériennes - Bacterial Toxins, Institut Pasteur [Paris], Centre de recherche du CEA/DSV/iBiTec-S/SIMOPRO, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), and SFET
Subjects: [SDV]Life Sciences [q-bio], [SDV.TOX]Life Sciences [q-bio]/Toxicology, ComputingMilieux_MISCELLANEOUS
Abstract: International audience
Published: 2020
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41. Selected Papers from the 26th Meeting of the French Society for Toxinology - Bioengineering of Toxins

Author: Popoff, Michel R., Evelyne BENOIT, Toxines bactériennes - Bacterial Toxins, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), ERL CNRS 9004, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), M.R. POPOFF & E. BENOIT (Eds), and Toxins
Subjects: Venon peptides, Bacterial toxins, [SDV]Life Sciences [q-bio], [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Neurotoxins, Animal toxins, Bioengineering, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Plant toxins, Neutralizing antibodies, Toxin inhibitors
Abstract: International audience; Toxins are very potent molecules responsible for moderate to highly severe diseases in man and animals. They are produced by a wide variety of organisms ranging from bacteria, to fungus, animals, and plants. However, toxins are also potent therapeutic drugs such as botulinum neurotoxins that have a large number of medical applications or peptide toxins, for example, conotoxins, used in the treatment of pain. In addition, toxins are very useful tools in cell biology that have revealed key physiological processes such as exocytosis, cytoskeleton organization, and the investigation of ionic channels. Modifications of toxins by bioengineering represent a current and important issue in the development of novel applications of toxins, including medical applications and tools for basic science, as well as the development of novel and efficient countermeasures (neutralizing antibodies and specific inhibitors). Recent progress shows that toxins can be modified for specific applications by increasing their activity or their cell specificity (for example, the development of toxins for the treatment of pain or the development of novel technological tools) or, in contrast, by decreasing their activity and use as vaccine or development of neutralizing antibodies and inhibitors.
Published: 2020

42. Shiga Toxin Uptake and Sequestration in Extracellular Vesicles Is Mediated by Its B-Subunit

Author: Willysson, Annie, Ståhl, Anne-lie, Gillet, Daniel, Barbier, Julien, Cintrat, Jean-Christophe, Chambon, Valérie, Billet, Anne, Johannes, Ludger, Karpman, Diana, BARBE, Sylviane, BLANC - Génétique chimique de la voie du transport rétrograde - - RETROscreen2011 - ANR-11-BSV2-0018 - BLANC - VALID, Appel à projets générique - Preuve de concept in vivo de molécules contre le syndrome hémolytique et urémique - - AntiHUS2014 - ANR-14-CE16-0004 - Appel à projets générique - VALID, Des cellules aux tissus: au croisement de la Physique et de la Biologie - - CelTisPhyBio2011 - ANR-11-LABX-0038 - LABX - VALID, Initiative d'excellence - Paris Sciences et Lettres - - PSL2010 - ANR-10-IDEX-0001 - IDEX - VALID, Endocytic Membrane Compartmentalization by Galectins - GALECTCOMPART - - EC:FP7:ERC2014-04-01 - 2019-03-31 - 340485 - VALID, Department of Pediatrics [Lund, Sweden] (Clinical Sciences), Lund University [Lund], Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Chimie biologique des membranes et ciblage thérapeutique (CBMCT - UMR 3666 / U1143), Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), ANR-19-CE13-0001-01, ANR-11-BSV2-0018,RETROscreen,Génétique chimique de la voie du transport rétrograde(2011), ANR-14-CE16-0004,AntiHUS,Preuve de concept in vivo de molécules contre le syndrome hémolytique et urémique(2014), ANR-11-LABX-0038,CelTisPhyBio,Des cellules aux tissus: au croisement de la Physique et de la Biologie(2011), ANR-10-IDEX-0001,PSL,Paris Sciences et Lettres(2010), European Project: 340485,EC:FP7:ERC,ERC-2013-ADG,GALECTCOMPART(2014), and Centre National de la Recherche Scientifique (CNRS)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)
Subjects: Erythrocytes, Time Factors, Trihexosylceramides, lcsh:R, Uterine Cervical Neoplasms, lcsh:Medicine, Receptors, Cell Surface, Shiga Toxin 1, Shiga toxin, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Article, Protein Subunits, Protein Transport, Humans, Female, HeLa cells, [SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, extracellular vesicles, red blood cells, globotriaosylceramide
Abstract: Shiga toxin (Stx)-stimulated blood cells shed extracellular vesicles (EVs) which can transfer the toxin to the kidneys and lead to hemolytic uremic syndrome. The toxin can be taken up by renal cells within EVs wherein the toxin is released, ultimately leading to cell death. The mechanism by which Stx is taken up, translocated, and sequestered in EVs was addressed in this study utilizing the B-subunit that binds to the globotriaosylceramide (Gb3) receptor. We found that Stx1B was released in EVs within minutes after stimulation of HeLa cells or red blood cells, detected by live cell imaging and flow cytometry. In the presence of Retro-2.1, an inhibitor of intracellular retrograde trafficking, a continuous release of Stx-positive EVs occurred. EVs from HeLa cells possess the Gb3 receptor on their membrane, and EVs from cells that were treated with a glycosylceramide synthase inhibitor, to reduce Gb3, bound significantly less Stx1B. Stx1B was detected both on the membrane and within the shed EVs. Stx1B was incubated with EVs derived from blood cells, in the absence of cells, and was shown to bind to, and be taken up by, these EVs, as demonstrated by electron microscopy. Using a membrane translocation assay we demonstrated that Stx1B was taken up by blood cell- and HeLa-derived EVs, an effect enhanced by chloropromazine or methyl-&szlig, cyclodextrin, suggesting toxin transfer within the membrane. This is a novel mechanism by which EVs derived from blood cells can sequester their toxic content, possibly to evade the host response.
Published: 2020
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43. Revisiting Old Ionophore Lasalocid as a Novel Inhibitor of Multiple Toxins

Author: Daniel Gillet, Jean-Christophe Cintrat, Emmanuel Lemichez, Julien Barbier, Yu Wu, Nassim Mahtal, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Service de Chimie Bio-Organique et de Marquage (SCBM), Toxines bactériennes - Bacterial Toxins, Institut Pasteur [Paris]-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)
Subjects: Health, Toxicology and Mutagenesis, [SDV]Life Sciences [q-bio], Bacterial Toxins, lcsh:Medicine, Exotoxins, Golgi Apparatus, Clostridium difficile toxin B, Endosomes, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], Toxicology, medicine.disease_cause, Protective Agents, Shiga Toxin 1, Article, Microbiology, 03 medical and health sciences, chemistry.chemical_compound, symbols.namesake, 0302 clinical medicine, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], medicine, Escherichia coli, Golgi, Pseudomonas exotoxin, Humans, Veterinary drug, Diphtheria Toxin, toxin, 030304 developmental biology, Lasalocid, Diphtheria toxin, 0303 health sciences, Ionophores, Toxin, lcsh:R, Biological Transport, Golgi apparatus, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, 3. Good health, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, chemistry, lasalocid, 030220 oncology & carcinogenesis, symbols, Lysosomes, Intracellular
Abstract: The ionophore lasalocid is widely used as a veterinary drug against coccidiosis. We found recently that lasalocid protects cells from two unrelated bacterial toxins, the cytotoxic necrotizing factor-1 (CNF1) from Escherichia. coli and diphtheria toxin. We evaluated lasalocid&rsquo, s capacity to protect cells against other toxins of medical interest comprising toxin B from Clostridium difficile, Shiga-like toxin 1 from enterohemorrhagic E. coli and exotoxin A from Pseudomonas aeruginosa. We further characterized the impact of lasalocid on the endolysosomal and the retrograde pathways and organelle integrity, especially the Golgi apparatus. We found that lasalocid protects cells from all toxins tested and impairs the drop of vesicular pH along the trafficking pathways that are required for toxin sorting and translocation to the cytoplasm. Lasalocid also has an impact on the cellular distribution of GOLPH4 and GOLPH2 Golgi markers. Other intracellular trafficking compartments positive for EEA1 and Rab9A display a modified cellular pattern. In conclusion, lasalocid protects cells from multiple deadly bacterial toxins by corrupting vesicular trafficking and Golgi stack homeostasis.
Published: 2020
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44. Gambierol enhances evoked quantal transmitter release and blocks a potassium current in motor nerve terminals of the mouse neuromuscular junction

Author: Jordi Molgó, Sébastien Schlumberger, Makoto Sasaki, Haruhiko Fuwa, Louzao, Carmen M., Botana, Luis M., Denis Servent, Evelyne BENOIT, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut des Neurosciences Paris-Saclay (NeuroPSI), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), Department of Biomolecular Sciences, Graduate School of Life Sciences, Tohoku University [Sendai], Departamento de Farmacología, Facultad de Veterinaria, Universidade de Santiago de Compostela [Spain] (USC ), SIMOPRO, Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Hess P. (Ed.), International Society for the Study of Harmful Algae (ISSHA) and Institut Francais de Recherche pour l'Exploitation de la Mer (Ifremer), in cooperation with the Intergovernmental Oceanographic Commission of the United Nations Educational, Scientific and Cultural Organization (IOC UNESCO), BENOIT, Evelyne, Hess P. (Ed.), International Society for the Study of Harmful Algae (ISSHA) and Institut Francais de Recherche pour l'Exploitation de la Mer (Ifremer), in cooperation with the Intergovernmental Oceanographic Commission of the United Nations Educational, Scientific and Cultural Organization (IOC UNESCO), and Universidad de Santiago de Compostela [Spain] (USC)
Subjects: [SDV.TOX] Life Sciences [q-bio]/Toxicology, Gambierol, potassium current, marine biotoxin, quantal transmitter release, [SDV.TOX]Life Sciences [q-bio]/Toxicology, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], nerve terminal
Abstract: In recent years, a great interest was developed to synthetize biologically-active natural products of marine origin. This is due to both their complex molecular structures and chemical diversity, and also to their unique biological activities. Among ladder-shaped toxins, gambierol, originally isolated from cultured Gambierdiscus toxicus dinoflagellate cells, together with ciguatoxins, has been successfully synthesized permitting detailed analyses of its mode and mechanism of action. Gambierol and analogs are known to inhibit some voltage-gated K+ (Kv) channel subtypes in various cell types. The aim of the present study was (i) to investigate whether gambierol has an action on quantal transmitter release evoked by nerve impulses and (ii) to determine whether Kv channels in motor nerve terminals of the mammalian neuromuscular junction are sensitive to the toxin action. Using electrophysiological techniques, the results obtained show that gambierol (2-20 nM) had no significant action on the resting membrane potential of mouse hemidiaphragm muscle fibers. In addition, spontaneous quantal transmitter release, measured by recording spontaneous miniature endplate potential frequency, remained unaffected by gambierol in resting neuromuscular junction. Gambierol (2 nM) increased about eight-fold the mean quantal content of evoked endplate potentials, as determined at individual junctions of the phrenic-hemidiaphragm preparation equilibrated in a low-Ca2+ and high-Mg2+ medium. The ability of gambierol to enhance quantal transmitter release was related to the reduction of a fast K+ current in nerve terminals. Overall, the present results show for the first time that gambierol enhances evoked quantal transmitter release in response to nerve stimuli, suggesting that it can be used to reverse pre- or post-synaptic neuromuscular blockade.
Published: 2020

45. Report from the 26th Meeting on Toxinology, 'Bioengineering of Toxins', Organized by the French Society of Toxinology (SFET) and Held in Paris, France, 4–5 December 2019

Author: Evelyne Benoit, Pascale Marchot, Sylvie Diochot, Michel R. Popoff, IPMC - Institut de pharmacologie moléculaire et cellulaire, Institut de pharmacologie moléculaire et cellulaire (IPMC), Centre National de la Recherche Scientifique (CNRS)-Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Université Côte d'Azur (UCA), Toxines bactériennes - Bacterial Toxins, Institut Pasteur [Paris], Centre de recherche du CEA/DSV/iBiTec-S/SIMOPRO, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Architecture et fonction des macromolécules biologiques (AFMB), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Centre National de la Recherche Scientifique (CNRS), Aix Marseille Université (AMU), Université Nice Sophia Antipolis (1965 - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Centre National de la Recherche Scientifique (CNRS)-Université Côte d'Azur (UCA), Institut Pasteur [Paris] (IP), SIMOPRO, and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
Subjects: 0303 health sciences, History, Toxinology, Health, Toxicology and Mutagenesis, [SDV]Life Sciences [q-bio], 030302 biochemistry & molecular biology, lcsh:R, Library science, lcsh:Medicine, Meeting Report, Toxicology, 03 medical and health sciences, n/a, [SDV.TOX]Life Sciences [q-bio]/Toxicology, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology
Abstract: This 26th edition of the annual Meeting on Toxinology (RT26) of the SFET (http://sfet.asso.fr/international) was held at the Institut Pasteur of Paris on 4–5 December 2019 [...]
Published: 2020
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46. Structural Studies based on two Lysine Dioxygenases with Distinct Regioselectivity Brings Insights Into Enzyme Specificity within the Clavaminate Synthase-Like Family

Author: Bastard, Karine, Isabet, Tatiana, Stura, Enrico A., Legrand, Pierre, Zaparucha, Anne, Génomique métabolique (UMR 8030), Genoscope - Centre national de séquençage [Evry] (GENOSCOPE), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS), Synchrotron SOLEIL (SSOLEIL), Centre National de la Recherche Scientifique (CNRS), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), We acknowledge synchrotron SOLEIL (Saint-Aubin, France) for provision of radiation facilities on the PROXIMA-1 and PROXIMA-2A beamlines, and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université d'Évry-Val-d'Essonne (UEVE)
Subjects: Models, Molecular, [SDV]Life Sciences [q-bio], lcsh:R, lcsh:Medicine, Crystallography, X-Ray, Flavobacterium, Article, Mixed Function Oxygenases, Protein Structure, Tertiary, Substrate Specificity, Actinobacteria, Bacterial Proteins, lcsh:Q, lcsh:Science
Abstract: International audience; Iron(II)/α-ketoacid-dependent oxygenases (αKAOs) are enzymes that catalyze the oxidation of unactivated C-H bonds, mainly through hydroxylation. Among these, those that are active towards amino-acids and their derivatives are grouped in the Clavaminate Synthase Like (CSL) family. CSL enzymes exhibit high regio-and stereoselectivities with strict substrate specificity. This study reports the structural elucidation of two new regiodivergent members, KDO1 and KDO5, active towards lysine, and the structural and computational analysis of the whole family through modelling and classification of active sites. The structures of KDO1 and KDO5 in complex with their ligands show that one exact position in the active site controls the regioselectivity of the reaction. Our results suggest that the substrate specificity and high stereoselectivity typical of this family is linked to a lid that closes up in order to form a sub-pocket around the side chain of the substrate. This dynamic lid is found throughout the family with varying sequence and length and is associated with a conserved stable dimeric interface. Results from this study could be a starting-point for exploring the functional diversity of the CSL family and direct in vitro screening in the search for new enzymatic activities. Enzymes derived from microorganisms are particularly appealing for the design of chemical processes that are environmentally friendly and sustainable 1. Enzymes catalyze interconversions of many functional groups with well-defined chemo-, regio-and stereoselectivities, resulting in high atom economy and reduced waste during synthesis. Over the past 25 years, the tremendous progress in molecular biology, bioinformatics and process optimization have lowered the biotechnological barriers that were limiting the industrial use of enzymes 2-4. Much research has looked for novel enzymes able to catalyze challenging reactions such as the functionalization of non-activated (i.e. not alpha to CC or C-heteroatom double bonds) C-H bonds. Such reactions still resist most efforts made by organic chemists. However, enzymes that can catalyze the formation of various C-heteroatom bonds have been found 5,6. We have focused our work on the iron(II)/α-KetoAcid-dependent Oxygenases (αKAOs), a mechanistically diverse superfamily of enzymes that use dioxygen as the oxidant to catalyze several reactions via C-H bond activation, mostly hydroxylation 7-9. These enzymes operate with three substrates, diox-ygen, α-ketoglutarate (α-KG) and a primary substrate. One oxygen atom is transferred to a primary substrate to give an oxidized product, and the second oxygen is given to the α-KG, which upon oxidation, forms succinate and CO 2 10,11,12. Many αKAOs are tailoring enzymes involved in secondary metabolite biosynthesis. Such enzymes, often found in bacteria, have been reported to hydroxylate the side chains of free amino acids and derivatives, or tether peptides in non-ribosomal peptide biosynthesis 13. Since hydroxylated amino acids contain several stereogenic centers, they are valuable chiral building blocks for fine chemical synthesis, which justifies the intensive effort
Published: 2018
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47. Late-Stage Isotopic Carbon Labeling of Pharmaceutically Relevant Cyclic Ureas Directly from CO2

Author: Christer Halldin, Olivier Loreau, Nathalie Camus, Bertrand Kuhnast, Antonio Del Vecchio, Magnus Schou, Davide Audisio, Pascal Kessler, Frédéric Taran, Kaisa Horkka, Fabien Caillé, Arnaud Chevalier, Laboratoire de Marquage au Carbone 14 (LMC), Service de Chimie Bio-Organique et de Marquage (SCBM), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Médicaments et Technologies pour la Santé (MTS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Imagerie Moléculaire in Vivo (IMIV - U1023 - ERL9218), Service Hospitalier Frédéric Joliot (SHFJ), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), LTMB équipe 2 (LTMB2), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), European Project: 675071,H2020,H2020-MSCA-ITN-2015,ISOTOPICS(2016), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Gasparini, Sylvaine, and ISOTOPIC LABELING FOR DRUG INNOVATION - ISOTOPICS - - H20202016-01-01 - 2019-12-31 - 675071 - VALID
Subjects: chemistry.chemical_classification, 010405 organic chemistry, Late-stage isotopic labeling, Carbon-13, Substrate (chemistry), chemistry.chemical_element, Heterocycles, General Chemistry, 010402 general chemistry, 01 natural sciences, Combinatorial chemistry, Catalysis, 0104 chemical sciences, Isotopic labeling, Carbon dioxide, chemistry, Isotopes of carbon, [CHIM] Chemical Sciences, carbon-14, [CHIM]Chemical Sciences, Compounds of carbon, Carbon-14, Reactivity (chemistry), carbon-11, Carbon
Abstract: International audience; A robust, click chemistry inspired procedure for radiolabeling of cyclic ureas was developed. This protocol, suitable for all carbon isotopes (11C, 13C, 14C), is based on the direct functionalization of carbon dioxide: the universal building block for carbon radiolabeling. The strategy is operationally simple, reproducible in different radiochemistry centers, exhibits a remarkably wide substrate scope with short reaction times, and demonstrates superior reactivity compared to previously reported systems. With this procedure, a variety of pharmaceuticals and an unprotected peptide were labeled with high radiochemical efficiency.
Published: 2018
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48. C-terminal truncation of IFN-γ inhibits proinflammatory macrophage responses and is deficient in autoimmune disease

Author: Jacob Rozmus, Antoine Dufour, Nestor Solis, Theo Klein, Vincent Dive, Nikolaus Fortelny, Caroline L. Bellac, Paul Pavlidis, Christopher M. Overall, Sean J. Barbour, Jennifer Mark, Ulrich Eckhard, Reinhild Kappelhoff, Parker G. Jobin, Department of Oral Biological and Medical Sciences, University of British Columbia (UBC), Centre for Blood Research (CBR), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Signalisation et physiopathologie cardiovasculaire (UMRS1180), and Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: Male, 0301 basic medicine, THP-1 Cells, Biopsy, [SDV]Life Sciences [q-bio], General Physics and Astronomy, Matrix metalloproteinase, Kidney, Mice, 0302 clinical medicine, Macrophage, THP1 cell line, skin and connective tissue diseases, lcsh:Science, Mice, Knockout, Multidisciplinary, biology, Chemistry, Lupus Nephritis, 3. Good health, Thioglycolates, Female, Collagen, medicine.symptom, Signal Transduction, Science, Primary Cell Culture, Inflammation, Matrix Metalloproteinase Inhibitors, Peritonitis, Article, General Biochemistry, Genetics and Molecular Biology, Autoimmune Diseases, Cell Line, Proinflammatory cytokine, Interferon-gamma, 03 medical and health sciences, Matrix Metalloproteinase 12, Synovitis, medicine, Animals, Humans, Autoimmune disease, MHC class II, Arthritis, Macrophages, General Chemistry, Macrophage Activation, medicine.disease, Mice, Inbred C57BL, Disease Models, Animal, RAW 264.7 Cells, 030104 developmental biology, Proteolysis, Immunology, biology.protein, lcsh:Q, 030217 neurology & neurosurgery
Abstract: Controlled macrophage differentiation and activation in the initiation and resolution of inflammation is crucial for averting progression to chronic inflammatory and autoimmune diseases. Here we show a negative feedback mechanism for proinflammatory IFN-γ activation of macrophages driven by macrophage-associated matrix metalloproteinase 12 (MMP12). Through C-terminal truncation of IFN-γ at 135Glu↓Leu136 the IFN-γ receptor-binding site was efficiently removed thereby reducing JAK-STAT1 signaling and IFN-γ activation of proinflammatory macrophages. In acute peritonitis this signature was absent in Mmp12–/– mice and recapitulated in Mmp12+/+ mice treated with a MMP12-specific inhibitor. Similarly, loss-of-MMP12 increases IFN-γ–dependent proinflammatory markers and iNOS+/MHC class II+ macrophage accumulation with worse lymphadenopathy, arthritic synovitis and lupus glomerulonephritis. In active human systemic lupus erythematosus, MMP12 levels were lower and IFN-γ higher compared to treated patients or healthy individuals. Hence, macrophage proteolytic truncation of IFN-γ attenuates classical activation of macrophages as a prelude for resolving inflammation., IFN-γ is central in inflammatory pathogenesis, response to infection and autoimmune diseases. Here the authors show that MMP12 expression is reduced in patients with SLE and that MMP12 post-translationally truncates IFN-y, inhibiting its function and affecting pathogenesis of mouse models of peritonitis, SLE and rheumatoid arthritis.
Published: 2018
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49. DMF-MALDI: droplet based microfluidic combined to MALDI-TOF for focused peptide detection

Author: Robert Thai, Sarah Bregant, Kiarach Mesbah, Florent Malloggi, Laboratoire Interdisciplinaire sur l'Organisation Nanométrique et Supramoléculaire (LIONS), Nanosciences et Innovation pour les Matériaux, la Biomédecine et l'Energie (ex SIS2M) (NIMBE UMR 3685), Institut Rayonnement Matière de Saclay (IRAMIS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Institut Rayonnement Matière de Saclay (IRAMIS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), ANR-10-NANO-02, CEA Program - Technologie pour la Santé, Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut Rayonnement Matière de Saclay (IRAMIS), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)
Subjects: 0301 basic medicine, Analyte, Science, Microfluidics, Sus scrofa, Peptide, Bioinformatics, 01 natural sciences, Article, 03 medical and health sciences, Protein sequencing, Animals, Humans, chemistry.chemical_classification, Multidisciplinary, Aqueous solution, Chromatography, Angiotensin II, 010401 analytical chemistry, [CHIM.MATE]Chemical Sciences/Material chemistry, Integrated approach, 0104 chemical sciences, 030104 developmental biology, chemistry, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Medicine, Sensitivity limit, Peptides
Abstract: We present an automated droplet microfluidic system (DMF) to generate monitored nanoliter aqueous droplets in oil and their deposition on a commercial stainless steel plate for MALDI-TOF analysis of peptides or protein digests. We demonstrate that DMF-MALDI combination focuses the analyte on the MALDI plate, increasing considerably the homogeneity of the dried material. This results in a 30times enhanced MALDI-TOF MS signal for a model peptide, allowing a significant improvement of the detection sensitivity limit (down to few tens of attomoles). Moreover, positive detection can be achieved from sub-nanomolar peptides solutions and better overall protein sequence coverages are obtained from few tens attomoles of protein digest. These results make DMF-MALDI a promising approach for the treatment of peptides samples as well as a key component for an integrated approach in the proteomic field.
Published: 2017
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50. Novel Matrix Metalloproteinase 12 Selective Radiotracers for Vascular Molecular Imaging

Author: Hye-Yeong Kim, Yunpeng Ye, Olivier Lequin, Jae-Joon Jung, Kiran Babu Gona, Vincent Dive, Fabrice Beau, Mahmoud Razavian, Isabelle Correia, Jakub Toczek, Mehran M. Sadeghi, Dimitris Georgiadis, Laurent Devel, Thomas Bordenave, Université Joseph Fourier - Grenoble 1 (UJF), Centre de recherche du CEA/DSV/iBiTec-S/SIMOPRO, Université Pierre et Marie Curie - Paris 6 (UPMC), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Yale School of Medicine [New Haven, Connecticut] (YSM), University of Athens, Athens, Laboratoire des biomolécules (LBM UMR 7203), Chimie Moléculaire de Paris Centre (FR 2769), École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Département de Chimie - ENS Paris, and Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Male, Biodistribution, [SDV.IB.IMA]Life Sciences [q-bio]/Bioengineering/Imaging, Matrix metalloproteinase, Matrix Metalloproteinase Inhibitors, 01 natural sciences, Article, 03 medical and health sciences, Aortic aneurysm, Mice, In vivo, medicine.artery, Matrix Metalloproteinase 12, Drug Discovery, medicine, Animals, Humans, [CHIM]Chemical Sciences, Tissue Distribution, Radioactive Tracers, Aorta, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Metalloproteinase, Radiochemistry, Chemistry, Organotechnetium Compounds, medicine.disease, Molecular biology, In vitro, 0104 chemical sciences, Molecular Imaging, Up-Regulation, Mice, Inbred C57BL, 010404 medicinal & biomolecular chemistry, cardiovascular system, Molecular Medicine, Molecular imaging, Aortic Aneurysm, Abdominal
Abstract: Matrix metalloproteinase-12 (MMP-12) is highly upregulated in several inflammatory diseases, including abdominal aortic aneurysm (AAA). Here we report four novel (99m)Tc-labeled radiotracers derived from a highly selective competitive MMP-12 inhibitor. These tracers in their (99g)Tc version were assessed in vitro on a set of human metalloproteases and displayed high affinity and selectivity toward MMP-12. Their radiolabeling with (99m)Tc was shown to be efficient and stable in both buffer and mouse blood. The tracers showed major differences in their biodistribution and blood clearance. Based on its in vivo performance, [(99m)Tc]-1 was selected for evaluation in murine AAA, where MMP-12 gene expression is upregulated. Autoradiography of aortae at two hours post-injection revealed high uptake of [(99m)Tc]-1 in AAA relative to adjacent aorta. Tracer uptake specificity was demonstrated through in vivo competition. This study paves the way for further evaluation of [(99m)Tc]-1 for imaging AAA and other MMP-12-associated diseases.
Published: 2019
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