Search

Your search keyword '"Theraptosis S.A."' showing total 23 results
Start Over Author "Theraptosis S.A."
23 results on '"Theraptosis S.A."'

1. Role of adherent and invasive E. coli in Crohn's Disease: Lessons from the postoperative recurrence model

Author

Buisson, Anthony, Sokol, Harry, Hammoudi, Nassim, Nancey, Stéphane, Treton, Xavier, Nachury, Maria, Fumery, Mathurin, Hebuterne, Xavier, Rodrigues, Michael, Hugot, Jean-Pierre, Boschetti, Gilles, Stefanescu, Carmen, Wils, Pauline, Seksik, Philippe, Le Bourhis, Lionel, Bezault, Madeleine, Sauvanet, Pierre, Pereira, Bruno, Allez, Matthieu, Barnich, N., CHU Clermont-Ferrand, Service de rhumatologie [CHU Saint-Antoine], CHU Saint-Antoine [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Microbes évolution phylogénie et infections (MEPHI), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Institut Hospitalier Universitaire Méditerranée Infection (IHU Marseille), Centre Hospitalier Lyon Sud [CHU - HCL] (CHLS), Hospices Civils de Lyon (HCL), Centre International de Recherche en Infectiologie - UMR (CIRI), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Hôpital Claude Huriez [Lille], CHU Lille, CHU Amiens-Picardie, Périnatalité et Risques Toxiques - UMR INERIS_I 1 (PERITOX), Institut National de l'Environnement Industriel et des Risques (INERIS)-Université de Picardie Jules Verne (UPJV)-CHU Amiens-Picardie, Département de Gastroentérologie, Hôpital de l'Archet 2, Beaujon, Assistance Publique - Hôpitaux de Paris, Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), Sorbonne Université (SU), Theraptosis S.A., Microbes, Intestin, Inflammation et Susceptibilité de l'Hôte (M2iSH), Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Recherche en Nutrition Humaine d'Auvergne (CRNH d'Auvergne)-Institut National de la Recherche Agronomique (INRA), Institut National de la Santé et de la Recherche Médicale (INSERM), Université Clermont Auvergne [2017-2020] (UCA [2017-2020]), Centre de Recherche en Nutrition Humaine, Service de chirurgie digestive, Hôpital Saint-André, Hôpital Saint-Louis, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Centre de Recherche en Nutrition Humaine d'Auvergne (CRNH d'Auvergne)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), DESSAIVRE, Louise, Service d'Hépatologie Gastro-entérologie [CHU Clermont-Ferrand], CHU Estaing [Clermont-Ferrand], CHU Clermont-Ferrand-CHU Clermont-Ferrand, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Service de Gastroentérologie [Hôpital Beaujon], Hôpital Beaujon [AP-HP], Servcie Hépato-gastroentérolgie et Nutrition clinique [CHU Nice], Centre Hospitalier Universitaire de Nice (CHU Nice), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Recherche en Nutrition Humaine d'Auvergne (CRNH d'Auvergne)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Clermont Auvergne (UCA), Centre de recherche sur l'Inflammation (CRI (UMR_S_1149 / ERL_8252 / U1149)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), 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), Hopital Saint-Louis [AP-HP] (AP-HP), Remind Study Group, Unité de Biostatistiques [CHU Clermont-Ferrand], Direction de la recherche clinique et de l’innovation [CHU Clermont-Ferrand] (DRCI), and European Crohn’s and Colitis Organisation

Subjects
[SDV] Life Sciences [q-bio], [SDV]Life Sciences [q-bio], [SDV.MHEP.HEG]Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, [SDV.MHEP.HEG] Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, ComputingMilieux_MISCELLANEOUS
Abstract

International audience

Published
2022

2. Long-term outcomes after ileocolonic resection in Crohn's disease according to the severity of early endoscopic recurrence

Author

Fumery, Mathurin, Le Bourhis, L., Nancey, S., Allez, M., Grp, Remind Study, Hammoudi, N., Auzolle, C., Minh, M. L. Tran, Boschetti, G., Bezault, M., Buisson, A., Pariente, B., Treton, X., Seksik, P., CHU Amiens-Picardie, Périnatalité et Risques Toxiques - UMR INERIS_I 1 (PERITOX), Institut National de l'Environnement Industriel et des Risques (INERIS)-Université de Picardie Jules Verne (UPJV)-CHU Amiens-Picardie, Centre Hospitalier Lyon Sud [CHU - HCL] (CHLS), Hospices Civils de Lyon (HCL), Hôpital Saint-Louis, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7), Microbes évolution phylogénie et infections (MEPHI), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Institut Hospitalier Universitaire Méditerranée Infection (IHU Marseille), Department of Gastroenterology, Centre Hospitalier Général Cannes, Theraptosis S.A., CHU Clermont-Ferrand, Hôpital Claude Huriez [Lille], CHU Lille, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), CHU Saint-Antoine [AP-HP], and Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)

Subjects
[SDV]Life Sciences [q-bio], ComputingMilieux_MISCELLANEOUS
Abstract

International audience

Published
2020

3. Osteoblastic differentiation and potent osteogenicity of three-dimensional hBMSC-BCP particle constructs

Author

Cordonnier, Thomas, Langonné, Alain, Corre, Pierre, Renaud, Audrey, Sensebé, Luc, Rosset, Philippe, Layrolle, Pierre, Sohier, Jérôme, Jehan, Frederic, Regenerating Bone defects using New biomedical Engineering approaches - REBORNE - - EC:FP7:HEALTH2010-01-01 - 2015-06-30 - 241879 - VALID, Physiopathologie des Adaptations Nutritionnelles (PhAN), Institut National de la Recherche Agronomique (INRA)-Université de Nantes (UN), Laboratoire d'hématopoïèse [Tours], Université Francois Rabelais [Tours], Etablissement Français du Sang Centre-Atlantique (EA3855), EFS, Regenerative Medicine and Skeleton (RMeS), École nationale vétérinaire, agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre hospitalier universitaire de Nantes (CHU Nantes)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN), Département de Stomatologie et chirurgie maxillo-faciale [CHU Nantes], Centre hospitalier universitaire de Nantes (CHU Nantes), Etablissement français du sang [Pyrénées-Méditerranée] (EFS Pyrénées-Méditerranée), Départements de chirurgie orthopédique [CHRU Tours], This work was supported by the French National Research Agency (ATOS project) and partially funded by the Directorate-General for Research of the European Commission (Grant No. 241879) through the REBORNE project., European Project: 241879,EC:FP7:HEALTH,FP7-HEALTH-2009-single-stage,REBORNE(2010), Micro et Nanomédecines Biomimétiques, Université d'Angers (UA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Bretagne Loire (UBL), Theraptosis Research Laboratory, Theraptosis S.A., Regenerative Medicine and Skeleton research lab (RMeS), Ecole Nationale Vétérinaire, Agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre hospitalier universitaire de Nantes (CHU Nantes)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Physiopathologie de la Résorption Osseuse et Thérapie des Tumeurs Osseuses Primitives, Université de Nantes (UN)-Institut National de la Santé et de la Recherche Médicale (INSERM), STROMALab, Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement Français du Sang-Centre National de la Recherche Scientifique (CNRS), Sarcomes osseux et remodelage des tissus calcifiés - Phy-Os [Nantes] (INSERM U1238), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Bretagne Loire (UBL)-Centre hospitalier universitaire de Nantes (CHU Nantes)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Equipe de recherche clinique en Odontologie (ERT1051), Université de Nantes (UN), Micro et Nanomédecines Biomimétiques (MINT), Ecole Nationale Vétérinaire, Agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS)-Centre hospitalier universitaire de Nantes (CHU Nantes)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement Français du Sang-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-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)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Centre Hospitalier Régional Universitaire de Tours (CHRU TOURS)-Université Francois Rabelais [Tours], and Centre Hospitalier Régional Universitaire de Tours (CHRU Tours)-Université Francois Rabelais [Tours]

Subjects
Male, three dimensional constructs, biphasic calcium phosphate, Mice, Nude, Real-Time Polymerase Chain Reaction, Mice, Implants, Experimental, Osteogenesis, Animals, Humans, RNA, Messenger, [SDV.IB.BIO]Life Sciences [q-bio]/Bioengineering/Biomaterials, bone tissue engineering, ComputingMilieux_MISCELLANEOUS, Aged, Cell Proliferation, Aged, 80 and over, [SDV.MHEP.RSOA] Life Sciences [q-bio]/Human health and pathology/Rhumatology and musculoskeletal system, Osteoblasts, Tissue Scaffolds, [SDV.MHEP.GEG] Life Sciences [q-bio]/Human health and pathology/Geriatry and gerontology, [SDV.MHEP.GEG]Life Sciences [q-bio]/Human health and pathology/Geriatry and gerontology, glass beads, Cell Differentiation, Mesenchymal Stem Cells, Middle Aged, Microspheres, osteogenicity in nude mice, human bone marrow stromal cells, [SDV.MHEP.RSOA]Life Sciences [q-bio]/Human health and pathology/Rhumatology and musculoskeletal system, Female, Hydroxyapatites, osteoblastic differentiation
Abstract

International audience; Bone tissue engineering usually consists of associating osteoprogenitor cells and macroporous scaffolds. This study investigated the in vitro osteoblastic differentiation and resulting in vivo bone formation induced by a different approach that uses particles as substrate for human bone marrow stromal cells (hBMSCs), in order to provide cells with a higher degree of freedom and allow them to synthesize a three-dimensional (3D) environment. Biphasic calcium phosphate (BCP) particles (35 mg, ~175 µm in diameter) were therefore associated with 4 X 10[5] hBMSCs. To discriminate the roles of BCP properties and cell-synthesized 3D environments, inert glass beads (GBs) of similar size were used under the same conditions. In both cases, high cell proliferation and extensive extracellular matrix (ECM) production resulted in the rapid formation of thick cell-synthesized 3D constructs. In vitro, spontaneous osteoblastic differentiation was observed in the 3D constructs at the mRNA and protein levels by monitoring the expression of Runx2, BMP2, ColI, BSP and OCN. The hBMSC–BCP particle constructs implanted in the subcutis of nude mice induced abundant ectopic bone formation after 8 weeks (~35%, n = 5/5). In comparison, only fibrous tissue without bone was observed in the implanted hBMSC–GB constructs (n = 0/5). Furthermore, little bone formation (~3%, n = 5/5) was found in hBMSC–macroporous BCP discs (diameter 8 X 3 mm). This study underlines the lack of correspondence between bone formation and in vitro differentiation assays. Furthermore, these results highlight the importance of using BCP as well as a 3D environment for achieving high bone yield of interest for bone engineering.

Published
2014
Full Text
View/download PDF

4. Consistent Osteoblastic Differentiation of Human Mesenchymal Stem Cells with Bone Morphogenetic Protein 4 and Low Serum

Author

Luc Sensebé, Frédéric Deschaseaux, Pierre Layrolle, Alain Langonné, Jérôme Sohier, Philippe Rosset, Thomas Cordonnier, Micro et Nanomédecines Biomimétiques, Université d'Angers (UA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Bretagne Loire (UBL), Theraptosis Research Laboratory, Theraptosis S.A., Physiopathologie de la Résorption Osseuse et Thérapie des Tumeurs Osseuses Primitives, Université de Nantes (UN)-Institut National de la Santé et de la Recherche Médicale (INSERM), Equipe de recherche clinique en Odontologie (ERT1051), Université de Nantes (UN), Sarcomes osseux et remodelage des tissus calcifiés - Phy-Os [Nantes] (INSERM U1238), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Bretagne Loire (UBL)-Centre hospitalier universitaire de Nantes (CHU Nantes)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN), Etablissement Français du Sang Centre-Atlantique (EA3855), EFS, Micro et Nanomédecines Biomimétiques (MINT), Physiopathologie des Adaptations Nutritionnelles (PhAN), Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Sarcomes osseux et remodelage des tissus calcifiés - Phy-Os [Nantes - INSERM U1238] (Phy-Os), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), and Université de Nantes (UN)-Université de Nantes (UN)-Centre hospitalier universitaire de Nantes (CHU Nantes)-Université Bretagne Loire (UBL)

Subjects
Adult, Serum, Ceramics, Biomedical Engineering, Medicine (miscellaneous), Bioengineering, Bone Morphogenetic Protein 4, Bone morphogenetic protein, Regenerative medicine, 03 medical and health sciences, Mice, 0302 clinical medicine, Osteogenesis, medicine, Animals, Humans, [SDV.IB.BIO]Life Sciences [q-bio]/Bioengineering/Biomaterials, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, Aged, Cell Proliferation, Aged, 80 and over, 0303 health sciences, Osteoblasts, Tissue Scaffolds, Chemistry, Gene Expression Profiling, Mesenchymal stem cell, Cell Differentiation, Mesenchymal Stem Cells, Middle Aged, Alkaline Phosphatase, Flow Cytometry, In vitro, 3. Good health, Cell biology, Bone morphogenetic protein 7, Bone morphogenetic protein 6, medicine.anatomical_structure, Bone morphogenetic protein 4, Gene Expression Regulation, 030220 oncology & carcinogenesis, Immunology, Bone marrow
Abstract

Providing fully mature and functional osteoblasts is challenging for bone tissue engineering and regenerative medicine. Such cells could be obtained from multipotent bone marrow mesenchymal stem cells (MSCs) after induction by different osteogenic factors. However, there are some discrepancies in results, notably due to the use of sera and to the type of osteogenic factor. In this study, we compared the osteogenic differentiation of bone marrow MSCs induced by dexamethasone (Dex) or bone morphogenetic proteins (BMPs) by assessing phenotypes in vitro and functional osteoblasts in vivo. Reducing the content of fetal calf serum from 10% to 2% significantly increased the mineral deposition and expression of osteoblastic markers during osteogenesis. In comparison to Dex condition, the addition of BMP4 greatly improved the differentiation of MSCs into fully mature osteoblasts as seen by high expression of Osterix. These results were confirmed in different supportive matrixes, plastic flasks, or biphasic calcium phosphate biomaterials. In contrast to Dex-derived osteoblasts, BMP4-derived osteoblasts from MSCs were significantly able to produce new bone in subcutis of nude mice in accordance with in vitro results. In conclusion, we describe a convenient ex vivo method to produce consistently mature functional osteoblasts from human MSCs with use of BMP4 and low serum.

Published
2011
Full Text
View/download PDF

5. Specific caspase inhibitor Q-VD-OPh prevents neonatal stroke in P7 rat: a role for gender

Author

Jean Mariani, Christiane Charriaut-Marlangue, Etienne Jacotot, Catherine Goyenvalle, Luc-Marie Joly, David Chauvier, Sylvain Renolleau, Sébastien Fau, Service de réanimation néonatale et pédiatrique [CHU Trousseau], Université Pierre et Marie Curie - Paris 6 (UPMC)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Trousseau [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU), Neurobiologie des processus adaptatifs (NPA), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), heraptosis Research Laboratory, Theraptosis S.A., CHU Charles Foix [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), and CHU Trousseau [APHP]

Subjects
MESH: Cell Death, Male, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Pharmacology, Biochemistry, MESH: Animals, Newborn, Amino Acid Chloromethyl Ketones, Brain ischemia, 0302 clinical medicine, MESH: Cerebrovascular Accident, MESH: Neurologic Examination, Medicine, MESH: Animals, MESH: In Situ Nick-End Labeling, Enzyme Inhibitors, Caspase, Neurologic Examination, MESH: Statistics, Nonparametric, 0303 health sciences, Cell Death, biology, Cytochrome c, Gender Identity, MESH: Gene Expression Regulation, Caspase Inhibitors, MESH: Gender Identity, Stroke, MESH: Enzyme Inhibitors, Quinolines, MESH: Quinolines, MESH: Rats, Caspase 1, Caspase 3, Neuroprotection, Statistics, Nonparametric, 03 medical and health sciences, Cellular and Molecular Neuroscience, In Situ Nick-End Labeling, Animals, Humans, Neonatal stroke, 030304 developmental biology, MESH: Caspases, MESH: Humans, business.industry, MESH: Amino Acid Chloromethyl Ketones, medicine.disease, MESH: Male, Rats, Animals, Newborn, Gene Expression Regulation, Apoptosis, Immunology, biology.protein, business, 030217 neurology & neurosurgery
Abstract

Hypoxia-ischaemia in the developing brain results in brain injury with prominent features of apoptosis. In the present study, a third generation dipeptidyl broad-spectrum caspase inhibitor, quinoline-Val-Asp(Ome)-CH2-O-phenoxy (Q-VD-OPh), was tested in a model of unilateral focal ischaemia with reperfusion in 7-day-old rats. Q-VD-OPh (1 mg/kg, i.p.) reduced cell death, resulting in significant neuroprotection at 48 h of recovery (infarct volume of 12.6 +/- 2.8 vs. 24.3 +/- 2.2%, p = 0.006). The neuroprotective effects observed at 48 h post-ischaemia hold up at 21 days of survival time and attenuate neurological dysfunction. Analysis by gender revealed that females were strongly protected (6.7 +/- 3.3%, p = 0.006), in contrast to males in which there was no significant effect, when Q-VD-OPh was given after clip removal on the left common carotid artery. Immunoblot analysis demonstrated that Q-VD-OPh inhibits caspase 3 cleavage into its p17 active form and caspase 1 up-regulation and cleavage in vivo. Following ischaemia in P7 rats, males and females displayed different time course and pattern of cytochrome c release and active p17 caspase 3 during the first 24 h of recovery. In contrast, no significant difference was observed for caspase 1 expression between genders. These results indicate that ischaemia activates caspases shortly after reperfusion and that the sex of the animal may strongly influences apoptotic pathways in the pathogenesis of neonatal brain injury. The specificity, effectiveness, and reduced toxicity of Q-VD-OPh may determine the potential use of peptide-derived irreversible caspase inhibitors as promising therapeutics.

Published
2007

6. Ryanodine receptor leak mediated by caspase-8 activation leads to left ventricular injury after myocardial ischemia-reperfusion

Author

Jean-Michel Rauzier, Stéphanie Roberge, Anne-Marie Lompré, Jérémy Fauconnier, Yassine Sassi, Jérôme Thireau, David Chauvier, Etienne Jacotot, Patrice Bideaux, Cécile Cassan, Albano C. Meli, Steven Reiken, Alexandre Marchand, Christine Crozier, Jian Shan, Alain Lacampagne, Andrew R. Marks, Physiologie & médecine expérimentale du Cœur et des Muscles [U 1046] (PhyMedExp), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Institute of pharmacology and toxicology, Technische Universität Munchen - Université Technique de Munich [Munich, Allemagne] (TUM), Department of Physiology & Cellular Biophysics, Columbia University [New York], Unité de Recherche sur les Maladies Cardiovasculaires, du Métabolisme et de la Nutrition = Institute of cardiometabolism and nutrition (ICAN), Université Pierre et Marie Curie - Paris 6 (UPMC)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU), Theraptosis Research Laboratory, Theraptosis S.A., Department of Reproductive Biology, Imperial College London, Columbia University College of Physicians and Surgeons, and Unité de Recherche sur les Maladies Cardiovasculaires, du Métabolisme et de la Nutrition = Research Unit on Cardiovascular and Metabolic Diseases (ICAN)

Subjects
Mitochondrial ROS, medicine.medical_specialty, Heart Ventricles, Diastole, Myocardial Reperfusion Injury, 030204 cardiovascular system & hematology, Ryanodine receptor 2, Rats, Inbred WKY, Fluorescence, Nitric oxide, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Internal medicine, medicine, [SDV.MHEP.PHY]Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], Animals, Myocardial infarction, Ventricular remodeling, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Caspase 8, Multidisciplinary, Ventricular Remodeling, Ryanodine receptor, business.industry, Tumor Necrosis Factor-alpha, Myocardium, Ryanodine Receptor Calcium Release Channel, Biological Sciences, medicine.disease, Phenanthridines, Rats, Enzyme Activation, chemistry, Cardiology, cardiovascular system, business, Reperfusion injury
Abstract

Myocardial ischemic disease is the major cause of death worldwide. After myocardial infarction, reperfusion of infracted heart has been an important objective of strategies to improve outcomes. However, cardiac ischemia/reperfusion (I/R) is characterized by inflammation, arrhythmias, cardiomyocyte damage, and, at the cellular level, disturbance in Ca 2+ and redox homeostasis. In this study, we sought to determine how acute inflammatory response contributes to reperfusion injury and Ca 2+ homeostasis disturbance after acute ischemia. Using a rat model of I/R, we show that circulating levels of TNF-α and cardiac caspase-8 activity were increased within 6 h of reperfusion, leading to myocardial nitric oxide and mitochondrial ROS production. At 1 and 15 d after reperfusion, caspase-8 activation resulted in S-nitrosylation of the RyR2 and depletion of calstabin2 from the RyR2 complex, resulting in diastolic sarcoplasmic reticulum (SR) Ca 2+ leak. Pharmacological inhibition of caspase-8 before reperfusion with Q-LETD-OPh or prevention of calstabin2 depletion from the RyR2 complex with the Ca 2+ channel stabilizer S107 (“rycal”) inhibited the SR Ca 2+ leak, reduced ventricular arrhythmias, infarct size, and left ventricular remodeling after 15 d of reperfusion. TNF-α–induced caspase-8 activation leads to leaky RyR2 channels that contribute to myocardial remodeling after I/R. Thus, early prevention of SR Ca 2+ leak trough normalization of RyR2 function is cardioprotective.

Published
2011

7. Targeting neonatal ischemic brain injury with a pentapeptide-based irreversible caspase inhibitor

Author

G Debret, Alain Lacampagne, Catherine I. Rousset, Pierre Gressens, S Holifanjaniaina, R Casimir, Etienne Jacotot, Xinhua Wang, M Bezault, D. Chauvier, J-M Rauzier, Jean Mariani, M Smirnova, Henrik Hagberg, Sylvain Renolleau, Leslie Schwendimann, Stefan Matecki, Ylva Carlsson, Christiane Charriaut-Marlangue, Eric Bernard, P Rustin, A-P Trichet, M Hébert, Johan Hoebeke, S Ankri, Theraptosis S.A., Neurobiologie des processus adaptatifs (NPA), Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC), Université Pierre et Marie Curie - Paris 6 (UPMC), Service de réanimation néonatale et pédiatrique [CHU Trousseau], Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-CHU Trousseau [APHP]-Sorbonne Université (SU), Physiopathologie, conséquences fonctionnelles et neuroprotection des atteintes du cerveau en développement, Université Paris Diderot - Paris 7 (UPD7)-IFR2-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Diderot - Paris 7 (UPD7), Centre for the developing brain, Imperial College London, Department of Physiology, University of Gothenburg (GU), Immunologie et chimie thérapeutiques (ICT), Cancéropôle du Grand Est-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Physique Théorique de la Matière Condensée (LPTMC), Department of Obstetrics & Gynaecology, Sahlgrenska University Hospital, Centre Hospitalier Régional Universitaire de Nîmes (CHRU Nîmes), Physiologie & médecine expérimentale du Cœur et des Muscles [U 1046] (PhyMedExp), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier), Hôpital Charles Foix (UEF), Hôpital Charles Foix Ivry/Seine, French Ministry of Research, Inserm, Foundation Grace de Monaco, ANVAR, MRC, Univesity Pari s7, Premup, Medical Research Council UK, MRC Sweden, ALF-LUA, Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), CHU Trousseau [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Sahlgrenska University Hospital [Gothenburg], Centre Hospitalier Universitaire de Nîmes (CHU Nîmes), Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), CHU Charles Foix [AP-HP], and Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)

Subjects
Cancer Research, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Immunology, Excitotoxicity, Ischemia, Apoptosis, ischemia, Pharmacology, Cysteine Proteinase Inhibitors, medicine.disease_cause, Pentapeptide repeat, Neuroprotection, 03 medical and health sciences, Cellular and Molecular Neuroscience, Mice, 0302 clinical medicine, In vivo, medicine, Animals, Caspase, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Binding Sites, biology, Safety pharmacology, Cytochromes c, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Cell Biology, medicine.disease, Caspase Inhibitors, caspase inhibitor, 3. Good health, Rats, perinatal brain damage, Disease Models, Animal, Neuroprotective Agents, Animals, Newborn, Caspases, Hypoxia-Ischemia, Brain, biology.protein, Quinolines, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, Original Article, neuroprotection, Oligopeptides, 030217 neurology & neurosurgery
Abstract

International audience; Brain protection of the newborn remains a challenging priority and represents a totally unmet medical need. Pharmacological inhibition of caspases appears as a promising strategy for neuroprotection. In a translational perspective, we have developed a pentapeptide-based group II caspase inhibitor, TRP601/ORPHA133563, which reaches the brain, and inhibits caspases activation, mitochondrial release of cytochrome c, and apoptosis in vivo. Single administration of TRP601 protects newborn rodent brain against excitotoxicity, hypoxia–ischemia, and perinatal arterial stroke with a 6-h therapeutic time window, and has no adverse effects on physiological parameters. Safety pharmacology investigations, and toxicology studies in rodent and canine neonates, suggest that TRP601 is a lead compound for further drug development to treat ischemic brain damage in human newborns.

Published
2011

8. Predicting sensorimotor and memory deficits after neonatal ischemic stroke with reperfusion in the rat

Author

Etienne Jacotot, David Chauvier, Madeleine Bezault, Valentine Bouet, Michel Boulouard, Sylvain Renolleau, Steve Ankri, Thomas Freret, Pascale Schumann-Bard, Groupe Mémoire et Plasticité comportementale (GMPc), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), CHU Trousseau [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Department of Reproductive Biology, Imperial College London, Theraptosis Research Laboratory, Theraptosis S.A., Normandie Université (NU), Université de Caen Basse-Normandie, Service de Réanimation Néonatale et Pédiatrique, Université Pierre et Marie Curie - Paris 6 (UPMC) - Assistance publique - Hôpitaux de Paris (AP-HP) - CHU Trousseau [APHP], and Université de Caen Basse-Normandie (UNICAEN)

Subjects
Male, Statistics as Topic, Ischemia, Posterior parietal cortex, Sensory system, Motor Activity, Somatosensory system, Lesion, 03 medical and health sciences, Behavioral Neuroscience, 0302 clinical medicine, Double-Blind Method, Predictive Value of Tests, Pregnancy, Glial Fibrillary Acidic Protein, Reflex, medicine, Avoidance Learning, Animals, Antigens, Rats, Wistar, Maze Learning, Stroke, Gait Disorders, Neurologic, 030304 developmental biology, 0303 health sciences, Analysis of Variance, Memory Disorders, [SCCO.NEUR]Cognitive science/Neuroscience, [SCCO.NEUR] Cognitive science/Neuroscience, Age Factors, Brain, Ectodysplasins, medicine.disease, Motor coordination, Rats, Disease Models, Animal, Gliosis, Animals, Newborn, Ischemic Attack, Transient, Phosphopyruvate Hydratase, Reperfusion, Exploratory Behavior, Female, Proteoglycans, medicine.symptom, Psychology, Neuroscience, 030217 neurology & neurosurgery
Abstract

International audience; Among experimental models of perinatal ischemic stroke, Renolleau's model mimics selected types of stroke at birth, including ischemia and reperfusion. However, its behavioural consequences on development have been poorly described. Here, ischemia-reperfusion was performed in 7-day-old Wistar rats. Between the ages of 9 and 40 days, sensorimotor and memory functions were assessed. The infarcted area was analysed by immunohistochemistry at 40 days of age. The remaining lesion was in the parietal cortex, in the form of a cone-shaped area. This area contained glial cells but neither neurons nor macrophages. Transient focal neonatal ischemia led to sensorimotor alterations in early adulthood, such as postural asymmetry, motor coordination and somatosensory deficits, and hyperactivity, as well as cognitive impairments, such as spatial reference memory deficits. Based on these results, we propose here a selection of behavioural tests that should constitute meaningful tools for assessing sensory and cognitive functions after experimental neonatal ischemic stroke.

Published
2010

9. 3D environment on human mesenchymal stem cells differentiation for bone tissue engineering

Author

Jérôme Sohier, Pierre Layrolle, Thomas Cordonnier, Luc Sensebé, Julien Gaillard, Philippe Rosset, Alain Langonné, Ecosystèmes montagnards (UR EMGR), Institut national de recherche en sciences et technologies pour l'environnement et l'agriculture (IRSTEA), Physiopathologie de la Résorption Osseuse et Thérapie des Tumeurs Osseuses Primitives, Université de Nantes (UN)-Institut National de la Santé et de la Recherche Médicale (INSERM), Models for the performance analysis and the control of networks (MAESTRO), Inria Sophia Antipolis - Méditerranée (CRISAM), Institut National de Recherche en Informatique et en Automatique (Inria)-Institut National de Recherche en Informatique et en Automatique (Inria), Theraptosis Research Laboratory, Theraptosis S.A., Etablissement Français du Sang Centre-Atlantique (EA3855), EFS, Université de Nantes - UFR Odontologie, Université de Nantes (UN), Physiopathologie des Adaptations Nutritionnelles (PhAN), Institut National de la Recherche Agronomique (INRA)-Université de Nantes (UN), Université de Nantes - UFR Odontologie (UFR Odonto), Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), STROMALab, Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), and Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement Français du Sang-Centre National de la Recherche Scientifique (CNRS)

Subjects
Calcium Phosphates, Bone sialoprotein, Sialoglycoproteins, Cellular differentiation, Biomedical Engineering, Biophysics, Bone Morphogenetic Protein 2, Bioengineering, Bone and Bones, Biomaterials, Extracellular matrix, 03 medical and health sciences, Imaging, Three-Dimensional, 0302 clinical medicine, Tissue engineering, Integrin-Binding Sialoprotein, Humans, [SDV.IB.BIO]Life Sciences [q-bio]/Bioengineering/Biomaterials, ComputingMilieux_MISCELLANEOUS, Cell Proliferation, 030304 developmental biology, 0303 health sciences, Osteoblasts, Tissue Engineering, biology, Reverse Transcriptase Polymerase Chain Reaction, Mesenchymal stem cell, Biomaterial, Cell Differentiation, Mesenchymal Stem Cells, Alkaline Phosphatase, Cell biology, Durapatite, biology.protein, Polystyrenes, Stem cell, 030217 neurology & neurosurgery, Biomedical engineering
Abstract

In this work a novel method was developed to create a three dimensional environment at a cellular level for bone tissue engineering. Biphasic calcium phosphate (BCP) particles of 140-200 microm were used in association with human mesenchymal stem cells (hMSCs). The cells seeded on these particles adhered and proliferated more rapidly in the first day of culture compared to culture on plastic. Analyses of hMSCs cultured without osteogenic factors on BCP particles revealed an abundant extracellular matrix production forming 3-dimensional (3D) hMSCs/BCP particles constructs after few days. Bone morphogenetic 2 (BMP-2), bone sialoprotein (BSP) and ALP gene expression using real time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) confirmed that expression profiles were modified by the culture substrate while the addition of osteogenic medium enhanced bone markers expression. These results indicate that BCP particles alone are able to induce an osteoblastic differentiation of hMSCs that might be of interest for bone tissue engineering.

Published
2010

10. A flavivirus protein M-derived peptide directly permeabilizes mitochondrial membranes, triggers cell death and reduces human tumor growth in nude mice

Author

Dominique Rebouillat, Olivier Chaloin, Nelly Buron, Sylviane Muller, Magali Brabant, Sylvie Dupont, Alain Langonne, Jean Paul Briand, David Chauvier, Catherine Brenner, Hervé Lecoeur, Myriam Lassalle, Mathieu Porceddu, Olivier Deas, Richard Casimir, Annie Borgne-Sanchez, Etienne Jacotot, Ludwig Baux, Theraptosis Research Laboratory, Theraptosis S.A., Immunologie et chimie thérapeutiques (ICT), Cancéropôle du Grand Est-Centre National de la Recherche Scientifique (CNRS), Department of Reproductive Biology, and Imperial College London

Subjects
Cancer Research, Programmed cell death, Amino Acid Motifs, Molecular Sequence Data, Clinical Biochemistry, Cell, Mice, Nude, Pharmaceutical Science, Peptide, [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], Biology, Mitochondrion, Permeability, Mice, Viral Proteins, 03 medical and health sciences, Transduction (genetics), 0302 clinical medicine, Cell Line, Tumor, medicine, Animals, Humans, Amino Acid Sequence, Cell Proliferation, 030304 developmental biology, Membrane Potential, Mitochondrial, Pharmacology, chemistry.chemical_classification, 0303 health sciences, Cell Death, Flavivirus, Biochemistry (medical), Cell Biology, Survival Analysis, Xenograft Model Antitumor Assays, Molecular biology, Mitochondria, Protein Structure, Tertiary, 3. Good health, Protein M, medicine.anatomical_structure, Ectodomain, chemistry, Apoptosis, 030220 oncology & carcinogenesis, Mitochondrial Membranes, tat Gene Products, Human Immunodeficiency Virus, Mitochondrial Swelling, Peptides
Abstract

International audience; Dengue viruses belong to the Flavivirus family and are responsible for hemorrhagic fever in Human. Dengue virus infection triggers apoptosis especially through the expression of the small membrane (M) protein. Using isolated mitochondria, we found that synthetic peptides containing the C-terminus part of the M ectodomain caused apoptosis-related mitochondrial membrane permeabilization (MMP) events. These events include matrix swelling and the dissipation of the mitochondrial transmembrane potential (DeltaPsi(m)). Protein M Flavivirus sequence alignments and helical wheel projections reveal a conserved distribution of charged residues. Moreover, when combined to the cell penetrating HIV-1 Tat peptide transduction domain (Tat-PTD), this sequence triggers a caspase-dependent cell death associated with DeltaPsi(m) loss and cytochrome c release. Mutational approaches coupled to functional screening on isolated mitochondria resulted in the selection of a protein M derived sequence containing nine residues with potent MMP-inducing properties on isolated mitochondria. A chimeric peptide composed of a Tat-PTD linked to the 9-mer entity triggers MMP and cell death. Finally, local administration of this chimeric peptide induces growth inhibition of xenograft prostate PC3 tumors in immuno-compromised mice, and significantly enhances animal survival. Together, these findings support the notion of using viral genomes as valuable sources to discover mitochondria-targeted sequences that may lead to the development of new anticancer compounds.

Published
2009

11. Broad-spectrum caspase inhibitors: from myth to reality?

Author

Etienne Jacotot, D Chauvier, S Ankri, Christiane Charriaut-Marlangue, R Casimir, Theraptosis Research Laboratory, Theraptosis S.A., Neurobiologie des processus adaptatifs (NPA), Université Pierre et Marie Curie - Paris 6 (UPMC) - Centre National de la Recherche Scientifique (CNRS), and Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Hydrocarbons, Fluorinated, MESH: Benzyl Compounds, MESH: Neurons, Computational biology, Amino Acid Chloromethyl Ketones, Mice, 03 medical and health sciences, Broad spectrum, 0302 clinical medicine, Benzyl Compounds, Animals, Humans, MESH: Animals, [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Enzyme Inhibitors, Molecular Biology, MESH: Mice, 030304 developmental biology, Neurons, 0303 health sciences, Caspase inhibitors, MESH: Caspases, MESH: Humans, Chemistry, [SCCO.NEUR]Cognitive science/Neuroscience, [SCCO.NEUR] Cognitive science/Neuroscience, MESH: Amino Acid Chloromethyl Ketones, Cell Biology, Caspase Inhibitors, Cathepsins, MESH: Hydrocarbons, Fluorinated, MESH: Enzyme Inhibitors, MESH: Cathepsins, Quinolines, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], 030217 neurology & neurosurgery, MESH: Quinolines
Published
2007

12. Targeted Vpr-derived Peptides Reach Mitochondria to Induce Apoptosis of aVb3-Expressing Endothelial Cells

Author

Pierre Rustin, Etienne Jacotot, Dominique Rebouillat, J. Brière, Hervé Lecoeur, Déas O, Deniaud A, Myriam Lassalle, Johan Hoebeke, Lena Edelman, Christine Péchoux, D. Chauvier, Magali Brabant, Roux P, Brenner C, Dupont S, Baux L, Alain Langonne, J. P. Briand, A. Borgne-Sanchez, Laboratoire de génétique et biologie cellulaire (LGBC), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Theraptosis Research Laboratory, Theraptosis S.A., Institut National de la Santé et de la Recherche Médicale (INSERM), Imagerie Dynamique (Plate-Forme) (PFID), Institut Pasteur [Paris], Unité de recherche génomique et physiologie de la lactation (GPL), Institut National de la Recherche Agronomique (INRA), Immunologie et chimie thérapeutiques (ICT), Cancéropôle du Grand Est-Centre National de la Recherche Scientifique (CNRS), Physiopathologie et neuroprotection des atteintes du cerveau en développement, Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM), Physiopathologie, conséquences fonctionnelles et neuroprotection des atteintes du cerveau en développement, Université Paris Diderot - Paris 7 (UPD7)-IFR2-Institut National de la Santé et de la Recherche Médicale (INSERM), Génomique et Physiologie de la Lactation (GPL), Centre National de la Recherche Scientifique (CNRS)-École pratique des hautes études (EPHE)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Institut Pasteur [Paris] (IP), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-École Pratique des Hautes Études (EPHE), and Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)

Subjects
[SDV]Life Sciences [q-bio], Apoptosis, MESH: Amino Acid Sequence, Mitochondrion, APOPTOSE, Mitochondrial apoptosis-induced channel, MESH: Dose-Response Relationship, Drug, Mice, 0302 clinical medicine, MESH: Mitochondrial Membranes, ENDOTHELIAL CELLS, BIOLOGIE CELLULAIRE, MESH: Animals, MESH: Endothelial Cells, Peptide sequence, Inbred BALB C, 0303 health sciences, Mice, Inbred BALB C, biology, MESH: Peptides, Cytochrome c, Adenine nucleotide translocator, PEPTIDES, Cell biology, Mitochondria, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], MESH: Integrin alphaVbeta3, MESH: Cell Survival, 030220 oncology & carcinogenesis, MESH: Permeability, Mitochondrial Membranes, Mitochondrial fission, Drug, CELLULAR TRAFFICKING APOPTOSIS, Voltage-dependent anion channel, MESH: Gene Products, vpr, MESH: Mitochondria, Cell Survival, Integrin, Molecular Sequence Data, MESH: Mice, Inbred BALB C, PTP, Permeability, Dose-Response Relationship, 03 medical and health sciences, Gene Products, Animals, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, [INFO]Computer Science [cs], Amino Acid Sequence, Molecular Biology, MESH: Mice, 030304 developmental biology, MESH: Molecular Sequence Data, MESH: Humans, Dose-Response Relationship, Drug, MESH: Apoptosis, Gene Products, vpr, vpr, Cell Biology, Integrin alphaVbeta3, biology.protein, Lysosomes, MESH: Lysosomes
Abstract

International audience; The HIV-1 encoded apoptogenic protein Vpr induces mitochondrial membrane permeabilization (MMP) via interactions with the voltage-dependent anion channel (VDAC) and the adenine nucleotide translocator (ANT). We have designed a peptide, TEAM-VP, composed of two functional domains, one a tumor blood vessel RGD-like 'homing' motif and the other an MMP-inducing sequence derived from Vpr. When added to isolated mitochondria, TEAM-VP interacts with ANT and VDAC, reduces oxygen consumption and overcomes Bcl-2 protection to cause inner and outer MMP. TEAM-VP specifically recognizes cell-surface expressed alpha(V)beta(3) integrins, internalizes, temporarily localizes to lysosomes and progressively co-distributes with the mitochondrial compartment with no sign of lysosomal membrane permeabilization. Finally TEAM-VP reaches mitochondria of angiogenic endothelial cells to induce mitochondrial fission, dissipation of the mitochondrial transmembrane potential (DeltaPsi(m)), cytochrome c release and apoptosis hallmarks. Hence, this chimeric peptide constitutes the first example of a virus-derived mitochondriotoxic compound as a candidate to kill selectively tumor neo-endothelia.

Published
2006

13. Peptido-targeting of the mitochondrial transition pore complex for therapeutic apoptosis induction

Author

Johan Hoebeke, Sylviane Muller, Catherine Brenner, Jean-Paul Briand, Aurélien Deniaud, Etienne Jacotot, Immunopathologie et immunointervention thérapeutique, Université Pierre et Marie Curie - Paris 6 (UPMC)-IFR58-Institut National de la Santé et de la Recherche Médicale (INSERM), Immunologie et chimie thérapeutiques (ICT), Cancéropôle du Grand Est-Centre National de la Recherche Scientifique (CNRS), Institut de biologie moléculaire et cellulaire (IBMC), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Institut de RadioAstronomie Millimétrique (IRAM), Centre National de la Recherche Scientifique (CNRS), Academia Sinica Institute of Astronomy and Astrophysics (ASIAA), Academia Sinica, Laboratoire Biodiversité et Fonctionnement des Ecosystèmes (LBFE), Université Paul Verlaine - Metz (UPVM), Laboratoire de Chimie et Applications (LCA), Theraptosis Research Laboratory, Theraptosis S.A., Laboratoire de génétique et biologie cellulaire (LGBC), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL), Université Pierre et Marie Curie - Paris 6 (UPMC) - IFR58 - Institut National de la Santé et de la Recherche Médicale (INSERM), Cancéropôle du Grand Est - Centre National de la Recherche Scientifique (CNRS), Institute of Astronomy and Astrophysics (ASIAA Sinica), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ) - École pratique des hautes études (EPHE) - Centre National de la Recherche Scientifique (CNRS), and Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)

Subjects
MESH: Signal Transduction, Pore complex, Aging, Apoptosis, Mitochondrion, Mitochondrial Membrane Transport Proteins, MESH: Neurodegenerative Diseases, 0302 clinical medicine, Drug Delivery Systems, Neoplasms, Drug Discovery, Homeostasis, MESH: Aging, MESH: Animals, MESH: Neoplasms, 0303 health sciences, MESH: Oxidative Stress, MESH: Peptides, Liver Diseases, Neurodegeneration, Neurodegenerative Diseases, MESH: Mitochondrial Membrane Transport Proteins, Cell biology, Mitochondria, MESH: Homeostasis, Reperfusion Injury, Intermembrane space, Signal Transduction, Programmed cell death, MESH: Liver Diseases, Stereochemistry, MESH: Mitochondria, MESH: Drug Delivery Systems, Biology, 03 medical and health sciences, medicine, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Animals, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, 030304 developmental biology, Pharmacology, MESH: Humans, Mitochondrial Permeability Transition Pore, MESH: Apoptosis, medicine.disease, Cytosol, Oxidative Stress, Mitochondrial permeability transition pore, MESH: Reperfusion Injury, Peptides, 030217 neurology & neurosurgery
Abstract

The permeability transition pore (PTPC), a polyprotein complex, participates in the mitochondrial homeostasis as well as in the mitochondrial phase of the intrinsic pathway of apoptosis. It integrates multiple death signals including alterations of the intracellular milieu, translocation of pro-apoptotic members of the Bax/Bcl-2 family, p53, and viral proteins. As a consequence, PTPC can act as a coordinator of the pro-apoptotic mitochondrial membrane permeabilization process and the release of pro-apoptotic intermembrane space proteins into the cytosol. Moreover, the deregulation of PTPC has been involved in several major human pathologies such as cancer, neurodegeneration, ischemia/reperfusion, aging, as well as hepatotoxicity. Therefore, PTPC has emerged as a promising potential therapeutic target. Here, we will review the current knowledge concerning the two opposite functions of the PTPC and its implication in various pathologies. We will discuss the possibility to target this complex with peptides to modulate apoptosis in an innovative therapeutic perspective.

Published
2006

14. N,N'-linked oligoureas as foldamers: chain length requirements for helix formation in protic solvent investigated by circular dichroism, NMR spectroscopy, and molecular dynamics

Author

Jean-Paul Briand, Richard Casimir, Vincent Semetey, Roland Graff, Christine Hemmerlin, Marie-Christine Averlant-Petit, Gilles Guichard, Aude Violette, Didier Rognan, Michel Marraud, Immunologie et chimie thérapeutiques (ICT), Cancéropôle du Grand Est-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie Physique Macromoléculaire (LCPM), Institut de Chimie du CNRS (INC)-Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS), Theraptosis Research Laboratory, Theraptosis S.A., Faculté de Chimie, Université Louis Pasteur - Strasbourg I, Institut Gilbert-Laustriat : Biomolécules, Biotechnologie, Innovation Thérapeutique, Centre National de la Recherche Scientifique (CNRS), ANR-05-BLAN-0119,SYNTEFOLDAME,Synthèse et étude structurale de foldamères pseudopeptidiques azotés(2005), Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), Cancéropôle du Grand Est - Centre National de la Recherche Scientifique (CNRS), Université de Lorraine (UL) - Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg - Centre National de la Recherche Scientifique (CNRS), ANR-05-BLAN-0119, SYNTEFOLDAME, Synthèse et étude structurale de foldamères pseudopeptidiques azotés(2005), Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS), Klotz, Evelyne, and Erhart, Marianne

Subjects
Models, Molecular, Circular dichroism, Stereochemistry, Molecular Conformation, MESH: Solvents, Nuclear Overhauser effect, 010402 general chemistry, 01 natural sciences, Biochemistry, Oligomer, Catalysis, MESH: Circular Dichroism, chemistry.chemical_compound, Colloid and Surface Chemistry, Tetramer, Biomimetic Materials, MESH: Nuclear Magnetic Resonance, Biomolecular, [CHIM.CHEM] Chemical Sciences/Cheminformatics, Side chain, Urea, Nuclear Magnetic Resonance, Biomolecular, MESH: Urea, MESH: Molecular Conformation, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], 010405 organic chemistry, MESH: Peptides, Circular Dichroism, Chemical shift, MESH: Biomimetic Materials, [SDV.BBM.BS] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], General Chemistry, Nuclear magnetic resonance spectroscopy, 0104 chemical sciences, [CHIM.THEO]Chemical Sciences/Theoretical and/or physical chemistry, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], [CHIM.THEO] Chemical Sciences/Theoretical and/or physical chemistry, chemistry, Solvents, Thermodynamics, MESH: Thermodynamics, Peptides, [CHIM.CHEM]Chemical Sciences/Cheminformatics, MESH: Models, Molecular, Protic solvent
Abstract

International audience; N,N'-linked oligoureas with proteinogenic side chains are peptide backbone mimetics belonging to the gamma-peptide lineage. In pyridine, heptamer 4 adopts a stable helical fold reminiscent of the 2.6(14) helical structure proposed for gamma-peptide foldamers. In the present study, we have used a combination of CD and NMR spectroscopies to correlate far-UV chiroptical properties and conformational preferences of oligoureas as a function of chain length from tetramer to nonamer. Both the intensity of the CD spectra and NMR chemical shift differences between alphaCH2 diastereotopic protons experienced a marked increase for oligomers between four and seven residues. No major change in CD spectra occurred between seven and nine residues, thus suggesting that seven residues could be the minimum length required for stabilizing a dominant conformation. Unexpectedly, in-depth NMR conformational investigation of heptamer 4 in CD3OH revealed that the 2.5 helix probably coexists with partially (un)folded conformations and that Z-E urea isomerization occurs, to some degree, along the backbone. Removing unfavorable electrostatic interactions at the amino terminal end of 4 and adding one H-bond acceptor by acylation with alkyl isocyanate (4 --> 7) was found to reinforce the 2.5 helical population. The stability of the 2.5 helical fold in MeOH is further discussed in light of unrestrained molecular dynamics (MD) simulation. Taken together, these new data provide additional insight into the folding propensity of oligoureas in protic solvent and should be of practical value for the design of helical bioactive oligoureas.

Published
2005

15. Upstream control of apoptosis by caspase-2 in serum-deprived primary neurons

Author

Jean-Claude Martinou, Dominique Rebouillat, D. Chauvier, Jean Mariani, Etienne Jacotot, Hervé Lecoeur, Alain Langonne, A. Borgne-Sanchez, Theraptosis Research Laboratory, Theraptosis S.A., Neurobiologie des processus adaptatifs (NPA), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), Department of Cell Biology, Sciences III, University of Geneva [Switzerland], Université Pierre et Marie Curie - Paris 6 (UPMC) - Centre National de la Recherche Scientifique (CNRS), and Université de Genève = University of Geneva (UNIGE)

Subjects
Serum, caspase-2, neuronal apoptosis, Cancer Research, Programmed cell death, Neurite, Cell Survival, Clinical Biochemistry, Caspase 2, Pharmaceutical Science, Apoptosis, Mitochondrion, serum deprivation, Models, Biological, Mice, 03 medical and health sciences, 0302 clinical medicine, checkpoint, Neurotrophic factors, Animals, Humans, Enzyme Inhibitors, Cells, Cultured, Caspase, bcl-2-Associated X Protein, 030304 developmental biology, Neurons, Pharmacology, 0303 health sciences, biology, Cytochrome c, Biochemistry (medical), Cytochromes c, [SDV.NEU.SC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Cognitive Sciences, Cell Biology, Cell biology, Enzyme Activation, mitochondria, Protein Transport, Bax, biology.protein, RNA Interference, Peptides, [SDV.NEU.SC] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Cognitive Sciences, 030217 neurology & neurosurgery
Abstract

During development as well as in pathological situations, neurons that fail to find appropriate targets or neurotrophic factors undergo cell death. Using primary cortical neurons subjected to acute serum-deprivation (SD), we have examined caspases activation, mitochondrial dysfunction and cell death parameters. Among a panel of metabolic, signaling and caspases inhibitors only those able to interfere with caspase-2 like activity protect primary neurons against SD-induced cell death. In situ detection and subcellular fractionation demonstrate a very early activation of cytosolic caspase-2, which controls Bax cleavage, relocalization and mitochondrial membrane permeabilization (MMP). Both z-VDVAD-fmk and a siRNA specific for caspase-2 abolish Bax changes, mitochondrial membranes permeabilization, as well as cytochrome c release-dependent activation of caspase-9/caspase-3, nuclear alterations, phosphatidylserine exposure, neurites dismantling and neuronal death. Hence, caspase-2 is an early checkpoint for apoptosis initiation in primary neurons subjected to serum deprivation.

Published
2005

16. Dynamic analysis of apoptosis in primary cortical neurons by fixed- and real-time cytofluorometry

Author

Hervé Lecoeur, Jean Mariani, Alain Langonne, Etienne Jacotot, Dominique Rebouillat, Bernard Brugg, D. Chauvier, Lena Edelman, Theraptosis Research Laboratory, Theraptosis S.A., Neurobiologie des processus adaptatifs (NPA), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), Technologie cellulaire anti-tumorale, Institut Pasteur [Paris] (IP), and Institut Pasteur [Paris]

Subjects
Cancer Research, cerebellum, Clinical Biochemistry, Pharmaceutical Science, Apoptosis, Mitochondrion, Flow cytometry, Membrane Potentials, cell counts, 03 medical and health sciences, chemistry.chemical_compound, Mice, 0302 clinical medicine, medicine, Fluorescence microscope, Animals, Caspase, 030304 developmental biology, Pharmacology, Membrane potential, Cerebral Cortex, Neurons, 0303 health sciences, biology, medicine.diagnostic_test, MPTP, naturally occurring cell death, Biochemistry (medical), [SDV.NEU.SC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Cognitive Sciences, Cell Biology, Flow Cytometry, Cell biology, Mitochondria, medicine.anatomical_structure, chemistry, biology.protein, Neuron, 030217 neurology & neurosurgery
Abstract

We describe here a cytofluorometric technology for the characterization of decision, execution, and degradation steps of neuronal apoptosis. Multiparametric flow cytometry was developped and combined to detailled fluorescence microscopy observations to establish the chronology and hierarchy of death-related events: neuron morphological changes, mitochondrial transmembrane potential (Δ Ψ m ) collapse, caspase-3 and -9 activation, phosphatidyl-serine exposure, nuclear dismantling and final plasma membrane permeabilization. Moreover, we developped a reliable real-time flow cytometric monitoring of Δ Ψ m and plasma membrane integrity in response to neurotoxic insults including MPTP treatment. Taking advantage of recently developped specific fluorescent probes and a third generation pan-caspase inhibitor, this integrated approach will be pertinent to study the cell biology of neuronal apoptosis and to characterize new neuro-toxic/protective molecules.

Published
2004

17. HIV-1 Tat protein directly induces mitochondrial membrane permeabilization and inactivates cytochrome c oxidase

Author

Olivier Chaloin, Etienne Daniel Francois Jacotot, Pierre Rustin, Mathieu Porceddu, Alain Langonne, Aurélien Deniaud, Magali Brabant, Catherine Brenner, Hervé Lecoeur, S. Muller, J-J Brière, J. P. Briand, Christine Péchoux, Nelly Buron, Dominique Rebouillat, Ralph El-Khoury, A. Borgne-Sanchez, Immunophysiologie et Parasitisme Intracellulaire, Institut Pasteur [Paris] (IP), Theraptosis S.A., Theraptosis SA, Mitologics SAS, Hôpital Robert Debré, Immunologie et chimie thérapeutiques (ICT), Cancéropôle du Grand Est-Centre National de la Recherche Scientifique (CNRS), Physiopathologie, conséquences fonctionnelles et neuroprotection des atteintes du cerveau en développement, Université Paris Diderot - Paris 7 (UPD7)-IFR2-Institut National de la Santé et de la Recherche Médicale (INSERM), Physiopathologie et neuroprotection des atteintes du cerveau en développement, Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM), Unité de recherche génomique et physiologie de la lactation (GPL), Institut National de la Recherche Agronomique (INRA), Laboratoire de génétique et biologie cellulaire (LGBC), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL), Signalisation et physiopathologie cardiaque, Université Paris-Sud - Paris 11 (UP11)-IFR141-Institut National de la Santé et de la Recherche Médicale (INSERM), INSERM UMR676, Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Laboratoire de Biochimie, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Robert Debré-Université Paris Diderot - Paris 7 (UPD7)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Robert Debré, Department of Reproductive Biology, Imperial College London, French Ministry of Research, ANVAR, Sidaction, AFM, Ammi, CNRS, Inserm, CRITT Ile de France, Institut Pasteur [Paris], CNRS FRE 2445, Centre National de la Recherche Scientifique (CNRS), Génomique et Physiologie de la Lactation (GPL), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Robert Debré-Université Paris Diderot - Paris 7 (UPD7)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Robert Debré, Pansiot, Sylvie, Laboratoire de génétique et biologie cellulaire, Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Centre National de la Recherche Scientifique (CNRS), and Jacotot, E

Subjects
Cancer Research, Cytochrome, Virologie, [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], Mitochondrion, 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid, medicine.disease_cause, MESH: 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid, Oxidative Phosphorylation, chemistry.chemical_compound, Mice, 0302 clinical medicine, membrane mitochondriale, MESH: Mitochondrial Membranes, MESH: Membrane Potential, Mitochondrial, cytochrome c oxidase, MESH: Animals, Membrane Potential, Mitochondrial, 0303 health sciences, Mice, Inbred BALB C, cytochrome c oxydase, biology, Cytochrome c, microscopie électronique, Brain, Cytochromes c, MESH: Cytochromes c, 3. Good health, Mitochondria, protéine virale, HIV-1, Tat, mitochondria, Liver, Proto-Oncogene Proteins c-bcl-2, mitochondrie, DIDS, MESH: Permeability, Mitochondrial Membranes, Original Article, tat Gene Products, Human Immunodeficiency Virus, MESH: Myocardium, Viral protein, MESH: Mitochondria, Immunology, MESH: Mice, Inbred BALB C, polarographie, potentiel transmembranaire, Oxidative phosphorylation, spectrophotométrie, spectrofluorométrie, Permeability, Electron Transport Complex IV, 03 medical and health sciences, Cellular and Molecular Neuroscience, MESH: Brain, MESH: Electron Transport Complex IV, MESH: Oxidative Phosphorylation, Virology, [SDV.BC.BC] Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], medicine, Cytochrome c oxidase, Animals, Humans, MESH: Mice, technique elisa, 030304 developmental biology, MESH: tat Gene Products, Human Immunodeficiency Virus, MESH: Humans, Ion Transport, synthèse de protéine, perméabilité cellulaire, Myocardium, Cell Biology, Molecular biology, MESH: Ion Transport, chemistry, MESH: Proto-Oncogene Proteins c-bcl-2, biology.protein, virus de l'immunodéficience humaine, 030217 neurology & neurosurgery, MESH: Liver
Abstract

International audience; The Trans-activator protein (Tat) of human immunodeficiency virus (HIV) is a pleiotropic protein involved in different aspects of AIDS pathogenesis. As a number of viral proteins Tat is suspected to disturb mitochondrial function. We prepared pure synthetic full-length Tat by native chemical ligation (NCL), and Tat peptides, to evaluate their direct effects on isolated mitochondria. Submicromolar doses of synthetic Tat cause a rapid dissipation of the mitochondrial transmembrane potential (ΔΨ(m)) as well as cytochrome c release in mitochondria isolated from mouse liver, heart, and brain. Accordingly, Tat decreases substrate oxidation by mitochondria isolated from these tissues, with oxygen uptake being initially restored by adding cytochrome c. The anion-channel inhibitor 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) protects isolated mitochondria against Tat-induced mitochondrial membrane permeabilization (MMP), whereas ruthenium red, a ryanodine receptor blocker, does not. Pharmacologic inhibitors of the permeability transition pore, Bax/Bak inhibitors, and recombinant Bcl-2 and Bcl-XL proteins do not reduce Tat-induced MMP. We finally observed that Tat inhibits cytochrome c oxidase (COX) activity in disrupted mitochondria isolated from liver, heart, and brain of both mouse and human samples, making it the first described viral protein to be a potential COX inhibitor.

Published
2012

18. Use of Human Cancer Cell Lines Mitochondria to Explore the Mechanisms of BH3 Peptides and ABT-737-Induced Mitochondrial Membrane Permeabilization

Author

Pierre Rustin, Christine Péchoux, Magali Brabant, Diana Desgué, Etienne Jacotot, Nelly Buron, Myriam Lassalle, Mathieu Porceddu, Annie Borgne-Sanchez, Cindy Racoeur, Theraptosis S.A., Mitologics SAS, Hôpital Robert Debré, Unité de recherche génomique et physiologie de la lactation (GPL), Institut National de la Recherche Agronomique (INRA), AP-HP Hôpital universitaire Robert-Debré [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Université Paris Diderot - Paris 7 (UPD7), Department of Reproductive Biology, and Imperial College London

Subjects
Non-Clinical Medicine/Research Methods, [SDV]Life Sciences [q-bio], Mitochondrion, Mitochondrial apoptosis-induced channel, Piperazines, Membrane Potentials, Nitrophenols, Mice, 0302 clinical medicine, BIOLOGIE CELLULAIRE, Inner mitochondrial membrane, Mice, Inbred BALB C, Sulfonamides, 0303 health sciences, Multidisciplinary, biology, Cytochrome c, Cell Biology/Cellular Death and Stress Responses, CANCER, Mitochondria, 3. Good health, Cell biology, PROTEINE BCL-2, Gene Expression Regulation, Neoplastic, Cross-Linking Reagents, Proto-Oncogene Proteins c-bcl-2, Biochemistry, mitochondrie, protéine, 030220 oncology & carcinogenesis, Mitochondrial Membranes, Medicine, Female, Biotechnology/Protein Chemistry and Proteomics, Biochemistry/Drug Discovery, biological phenomena, cell phenomena, and immunity, Bacterial outer membrane, Research Article, Science, Oncology/Oncology Agents, Permeability, 03 medical and health sciences, Cell Line, Tumor, Proto-Oncogene Proteins, Animals, Humans, [INFO]Computer Science [cs], 030304 developmental biology, apoptose, Biphenyl Compounds, Peptide Fragments, CYTOCHROME-C RELEASE, BCL-2 FAMILY PROTEINS, MIMETIC ABT-737, OUTER-MEMBRANE, CASPASE ACTIVATION, BAX, APOPTOSIS, DEATH, INHIBITORS, DOMAINS, membrane cellulaire, Apoptosis, Cancer cell, biology.protein, Apoptosome, humain
Abstract

International audience; Current limitations of chemotherapy include toxicity on healthy tissues and multidrug resistance of malignant cells. A number of recent anti-cancer strategies aim at targeting the mitochondrial apoptotic machinery to induce tumor cell death. In this study, we set up protocols to purify functional mitochondria from various human cell lines to analyze the effect of peptidic and xenobiotic compounds described to harbour either Bcl-2 inhibition properties or toxic effects related to mitochondria. Mitochondrial inner and outer membrane permeabilization were systematically investigated in cancer cell mitochondria versus non-cancerous mitochondria. The truncated (t-) Bid protein, synthetic BH3 peptides from Bim and Bak, and the small molecule ABT-737 induced a tumor-specific and OMP-restricted mitochondrio-toxicity, while compounds like HA-14.1, YC-137, Chelerythrine, Gossypol, TW-37 or EM20-25 did not. We found that ABT-737 can induce the Bax-dependent release of apoptotic proteins (cytochrome c, Smac/Diablo and Omi/HtrA2 but not AIF) from various but not all cancer cell mitochondria. Furthermore, ABT-737 addition to isolated cancer cell mitochondria induced oligomerization of Bax and/or Bak monomers already inserted in the mitochondrial membrane. Finally immunoprecipatations indicated that ABT-737 induces Bax, Bak and Bim desequestration from Bcl-2 and Bcl-xL but not from Mcl-1L. This study investigates for the first time the mechanism of action of ABT-737 as a single agent on isolated cancer cell mitochondria. Hence, this method based on MOMP (mitochondrial outer membrane permeabilization) is an interesting screening tool, tailored for identifying Bcl-2 antagonists with selective toxicity profile against cancer cell mitochondria but devoid of toxicity against healthy mitochondria.

Published
2010

19. Targeted Vpr-derived peptides reach mitochondria to induce apoptosis of alphaVbeta3-expressing endothelial cells.

Author

Borgne-Sanchez A, Dupont S, Langonné A, Baux L, Lecoeur H, Chauvier D, Lassalle M, Déas O, Brière JJ, Brabant M, Roux P, Péchoux C, Briand JP, Hoebeke J, Deniaud A, Brenner C, Rustin P, Edelman L, Rebouillat D, and Jacotot E

Subjects
Amino Acid Sequence, Animals, Apoptosis, Cell Survival, Dose-Response Relationship, Drug, Endothelial Cells metabolism, Gene Products, vpr pharmacology, Humans, Lysosomes metabolism, Mice, Mice, Inbred BALB C, Mitochondrial Membranes metabolism, Molecular Sequence Data, Peptides pharmacology, Permeability, Endothelial Cells physiology, Gene Products, vpr pharmacokinetics, Integrin alphaVbeta3 metabolism, Mitochondria metabolism, Peptides pharmacokinetics
Abstract

The HIV-1 encoded apoptogenic protein Vpr induces mitochondrial membrane permeabilization (MMP) via interactions with the voltage-dependent anion channel (VDAC) and the adenine nucleotide translocator (ANT). We have designed a peptide, TEAM-VP, composed of two functional domains, one a tumor blood vessel RGD-like 'homing' motif and the other an MMP-inducing sequence derived from Vpr. When added to isolated mitochondria, TEAM-VP interacts with ANT and VDAC, reduces oxygen consumption and overcomes Bcl-2 protection to cause inner and outer MMP. TEAM-VP specifically recognizes cell-surface expressed alpha(V)beta(3) integrins, internalizes, temporarily localizes to lysosomes and progressively co-distributes with the mitochondrial compartment with no sign of lysosomal membrane permeabilization. Finally TEAM-VP reaches mitochondria of angiogenic endothelial cells to induce mitochondrial fission, dissipation of the mitochondrial transmembrane potential (DeltaPsi(m)), cytochrome c release and apoptosis hallmarks. Hence, this chimeric peptide constitutes the first example of a virus-derived mitochondriotoxic compound as a candidate to kill selectively tumor neo-endothelia.

Published
2007
Full Text
View/download PDF

20. Upstream control of apoptosis by caspase-2 in serum-deprived primary neurons.

Author

Chauvier D, Lecoeur H, Langonné A, Borgne-Sanchez A, Mariani J, Martinou JC, Rebouillat D, and Jacotot E

Subjects
Animals, Caspase 2 deficiency, Cell Survival drug effects, Cells, Cultured, Cytochromes c metabolism, Enzyme Activation drug effects, Enzyme Inhibitors pharmacology, Humans, Mice, Mitochondria drug effects, Mitochondria enzymology, Mitochondria metabolism, Models, Biological, Neurons drug effects, Peptides pharmacology, Protein Transport drug effects, RNA Interference, bcl-2-Associated X Protein metabolism, Apoptosis drug effects, Caspase 2 metabolism, Neurons cytology, Neurons enzymology, Serum
Abstract

During development as well as in pathological situations, neurons that fail to find appropriate targets or neurotrophic factors undergo cell death. Using primary cortical neurons subjected to acute serum-deprivation (SD), we have examined caspases activation, mitochondrial dysfunction and cell death parameters. Among a panel of metabolic, signaling and caspases inhibitors only those able to interfere with caspase-2 like activity protect primary neurons against SD-induced cell death. In situ detection and subcellular fractionation demonstrate a very early activation of cytosolic caspase-2, which controls Bax cleavage, relocalization and mitochondrial membrane permeabilization (MMP). Both z-VDVAD-fmk and a siRNA specific for caspase-2 abolish Bax changes, mitochondrial membranes permeabilization, as well as cytochrome c release-dependent activation of caspase-9/caspase-3, nuclear alterations, phosphatidylserine exposure, neurites dismantling and neuronal death. Hence, caspase-2 is an early checkpoint for apoptosis initiation in primary neurons subjected to serum deprivation.

Published
2005
Full Text
View/download PDF

21. Real-time flow cytometry analysis of permeability transition in isolated mitochondria.

Author

Lecoeur H, Langonné A, Baux L, Rebouillat D, Rustin P, Prévost MC, Brenner C, Edelman L, and Jacotot E

Subjects
Alamethicin pharmacology, Animals, Benzimidazoles, Calcium pharmacology, Carbocyanines, Cell Membrane Permeability, Fluorescent Dyes, Intracellular Membranes metabolism, Kinetics, Membrane Potentials, Mice, Mice, Inbred BALB C, Mitochondria drug effects, Mitochondrial Membrane Transport Proteins, Mitochondrial Permeability Transition Pore, Flow Cytometry methods, Ion Channels metabolism, Mitochondria physiology, Mitochondria ultrastructure
Abstract

Mitochondrial membrane permeabilization (MMP) is a key event in necrotic and (intrinsic) apoptotic processes. MMP is controlled by a few major rate-limiting events, one of which is opening of the permeability transition pore (PTP). Here we develop a flow cytometry (FC)-based approach to screen and study inducers and blockers of MMP in isolated mitochondria. Fixed-time and real-time FC permits to co-evaluate and order modifications of mitochondrial size, structure and inner membrane (IM) electrochemical potential (DeltaPsi(m)) during MMP. Calcium, a major PTP opener, and alamethicin, a PTP-independent MMP inducer, trigger significant mitochondrial forward scatter (FSC) increase and side scatter (SSC) decrease, correlating with spectrophotometrically detected swelling. FC-based fluorescence detection of the DeltaPsi(m)-sensitive cationic lipophilic dye JC-1 permits to detect DeltaPsi(m) variations induced by PTP openers or specific inducers of inner MMP such as carbonylcyanide m-chlorophenylhydrazone (mClCCP). These simple, highly sensitive and quantitative FC-based methods will be pertinent to evaluate compounds for their ability to control MMP.

Published
2004
Full Text
View/download PDF

22. Dynamic analysis of apoptosis in primary cortical neurons by fixed- and real-time cytofluorometry.

Author

Lecoeur H, Chauvier D, Langonné A, Rebouillat D, Brugg B, Mariani J, Edelman L, and Jacotot E

Subjects
Animals, Cerebral Cortex cytology, Membrane Potentials physiology, Mice, Mitochondria physiology, Apoptosis physiology, Cerebral Cortex physiology, Flow Cytometry, Neurons physiology
Abstract

We describe here a cytofluorometric technology for the characterization of decision, execution, and degradation steps of neuronal apoptosis. Multiparametric flow cytometry was developed and combined to detailed fluorescence microscopy observations to establish the chronology and hierarchy of death-related events: neuron morphological changes, mitochondrial transmembrane potential (DeltaPsi(m)) collapse, caspase-3 and -9 activation, phosphatidyl-serine exposure, nuclear dismantling and final plasma membrane permeabilization. Moreover, we developed a reliable real-time flow cytometric monitoring of DeltaPsi(m) and plasma membrane integrity in response to neurotoxic insults including MPTP treatment. Taking advantage of recently developed specific fluorescent probes and a third generation pan-caspase inhibitor, this integrated approach will be pertinent to study the cell biology of neuronal apoptosis and to characterize new neuro-toxic/protective molecules., (Copyright 2004 Kluwer Academic Publishers)

Published
2004
Full Text
View/download PDF

23. Nuclear apoptosis detection by flow cytometry: influence of endogenous endonucleases.

Author

Lecoeur H

Subjects
Animals, Cell Nucleus, DNA Fragmentation, Enzyme Activation, Flow Cytometry methods, Humans, Apoptosis, Endonucleases metabolism
Abstract

Nuclear apoptosis is characterized by chromatin condensation and progressive DNA cleavage into high-molecular-weight fragments and oligonucleosomes. These complex phenomena can be mediated by the activation of a multiplicity of enzymes, characterized by specific patterns of cation dependance, pH requirement, and mode of activation. The significance of this multiplicity of enzymes that cleave genomic DNA has been attributed to the need of death effector pathways specific for cell types/tissues, the level of cell differenciation, and the nature of the apoptotic stimuli. The activation of these factors contributes to the development of alterations that can be detected specifically by flow cytometric assays, namely, propidium iodide assays, acridine orange/ethidium bromide double staining, the TUNEL and ISNT techniques, and the assays of DNA sensitivity to denaturation. Although applicable to a wide spectrum of cell types, an increasing body of literature indicates that these techniques cannot be universally applied to all cell lines and apoptotic conditions: The requirement of a particular mediator(s) of nuclear apoptosis or the absence of endonuclease activity can limit the relevance of certain techniques. Finally, endonucleases recruited during primary necrosis can introduce nuclear alterations detected by some assays and raise the problem of their specificity. This review underlines the need for strategies to accurately detect and quantify nuclear apoptosis by flow cytometry when new cell systems and apoptotic conditions are considered., ((c) 2002 Elsevier Science (USA).)

Published
2002
Full Text
View/download PDF

Catalog

Books, media, physical & digital resources
See catalog results