53 results on '"Arlette Hammann"'
Search Results
2. Serpin B1 defect and increased apoptosis of neutrophils in Cohen syndrome neutropenia
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Laurence Duplomb, Jean-Noël Bastie, Edward Blair, Julie Riviere, Laurent Delva, Romain Da Costa, Arlette Hammann, Julien Thevenon, Bernard Aral, Jamal Ghoumid, Marie-Anne Gougerot-Pocidalo, Arnaud Lafon, Christel Thauvin-Robinet, Alain Schmitt, Patrick Edery, Virginie Carmignac, Julien Guy, Nathalie Droin, Eric Solary, Laurence Faivre, Gaëtan Jego, Laurence Dubrez, Jessica Racine, Salima El Chehadeh-Djebbar, François Girodon, and Claude Capron
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Adult ,Male ,Programmed cell death ,Neutropenia ,Adolescent ,Neutrophils ,Developmental Disabilities ,Down-Regulation ,Apoptosis ,Fingers ,Young Adult ,03 medical and health sciences ,0302 clinical medicine ,Intellectual Disability ,Drug Discovery ,Myopia ,Humans ,Medicine ,Obesity ,Child ,Serpins ,Genetics (clinical) ,Cohen syndrome ,biology ,business.industry ,Retinal Degeneration ,SERPINB1 ,Middle Aged ,medicine.disease ,VPS13B ,Child, Preschool ,Neutrophil elastase ,Mutation ,Immunology ,Microcephaly ,biology.protein ,Unfolded protein response ,Muscle Hypotonia ,Molecular Medicine ,Female ,business ,030215 immunology - Abstract
Cohen syndrome (CS) is a rare genetic disorder due to mutations in VPS13B gene. Among various clinical and biological features, CS patients suffer from inconsistent neutropenia, which is associated with recurrent but minor infections. We demonstrate here that this neutropenia results from an exaggerate rate of neutrophil apoptosis. Besides this increased cell death, which occurs in the absence of any endoplasmic reticulum stress or defect in neutrophil elastase (ELANE) expression or localization, all neutrophil functions appeared to be normal. We showed a disorganization of the Golgi apparatus in CS neutrophils precursors, that correlates with an altered glycosylation of ICAM-1 in these cells, as evidenced by a migration shift of the protein. Furthermore, a striking decrease in the expression of SERPINB1 gene, which encodes a critical component of neutrophil survival, was detected in CS neutrophils. These abnormalities may account for the excessive apoptosis of neutrophils leading to neutropenia in CS. KEY MESSAGES: Cohen syndrome patients' neutrophils display normal morphology and functions. Cohen syndrome patients' neutrophils have an increased rate of spontaneous apoptosis compared to healthy donors' neutrophils. No ER stress or defective ELA2 expression or glycosylation was observed in Cohen syndrome patients' neutrophils. SerpinB1 expression is significantly decreased in Cohen syndrome neutrophils as well as in VPS13B-deficient cells.
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- 2019
3. Gap junction-mediated transfer of miR-145-5p from microvascular endothelial cells to colon cancer cells inhibits angiogenesis
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Gaëtan Jego, Arlette Hammann, Eric Solary, Carmen Garrido, Kevin Berthenet, Dominique Thuringer, Lipides - Nutrition - Cancer (U866) ( LNC ), Université de Bourgogne ( UB ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon ( ENSBANA ), Hématopoïèse normale et pathologique ( U1170 Inserm ), Université Paris-Sud - Paris 11 ( UP11 ) -Institut Gustave Roussy ( IGR ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc ( CRLCC - CGFL ), Centre National de la Recherche Scientifique (CNRS), Institut National de la Sante et de la Recherche Medicale (INSERM), Ligue Nationale Contre le Cancer, Agence Nationale de la Recherche, Institut National du Cancer (INCa), Lipides - Nutrition - Cancer (U866) (LNC), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA), Hématopoïèse normale et pathologique (U1170 Inserm), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] (UNICANCER/CRLCC-CGFL), and UNICANCER
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0301 basic medicine ,Cell signaling ,Angiogenesis ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,Cell Communication ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,[ SDV.CAN ] Life Sciences [q-bio]/Cancer ,03 medical and health sciences ,Cell Line, Tumor ,microRNA ,Bystander effect ,Humans ,Medicine ,Neovascularization, Pathologic ,[ SDV.BC ] Life Sciences [q-bio]/Cellular Biology ,business.industry ,micro-RNA ,Gap junction ,Endothelial Cells ,Gap Junctions ,tubulogenesis ,Coculture Techniques ,Microvesicles ,Cx43 ,Cell biology ,Endothelial stem cell ,MicroRNAs ,030104 developmental biology ,Oncology ,GJIC ,Colonic Neoplasms ,Cancer cell ,Immunology ,business ,Research Paper ,colorectal tumor - Abstract
IF 5.008; International audience; Gap junctional communication between cancer cells and blood capillary cells is crucial to tumor growth and invasion. Gap junctions may transfer microRNAs (miRs) among cells. Here, we explore the impact of such a transfer in co-culture assays, using the antitumor miR-145 as an example. The SW480 colon carcinoma cells form functional gap junction composed of connexin-43 (Cx43) with human microvascular endothelial cells (HMEC). When HMEC are loaded with miR-145-5p mimics, the miR-145 level drastically increases in SW480. The functional inhibition of gap junctions, using either a gap channel blocker or siRNA targeting Cx43, prevents this increase. The transfer of miR-145 also occurs from SW480 to HMEC but not in non-contact co-cultures, excluding the involvement of soluble exosomes. The miR-145 transfer to SW480 up-regulates their Cx43 expression and inhibits their ability to promote angiogenesis. Our results indicate that the gap junctional communication can inhibit tumor growth by transferring miRs from one endothelial cell to neighboring tumor cells. This "bystander" effect could find application in cancer therapy.
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- 2016
4. HSP110 promotes colorectal cancer growth through STAT3 activation
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Gaëtan Jego, Oleg N. Demidov, Kevin Berthenet, Anaïs Lagrange, Anastasia R. Goloudina, Alex Duval, A’dem Bokhari, Arlette Hammann, Christophe Boudesco, Ada Collura, Carmen Garrido, Sebastien Causse, S Dumont, A de Thonel, Denis Biard, Renaud Seigneuric, Magali Svrcek, Guillaume Marcion, Lipides - Nutrition - Cancer (U866) (LNC), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Bourgogne (UB)-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement, Centre de Recherche Saint-Antoine (UMRS893), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre épigénétique et destin cellulaire (EDC (UMR_7216)), Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA), Centre de Recherche Saint-Antoine (CR Saint-Antoine), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Centre épigénétique et destin cellulaire (EDC), Lipides - Nutrition - Cancer (U866) ( LNC ), Université de Bourgogne ( UB ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon ( ENSBANA ), Centre de Recherche Saint-Antoine ( CR Saint-Antoine ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Centre épigénétique et destin cellulaire ( EDC ), and Université Paris Diderot - Paris 7 ( UPD7 ) -Centre National de la Recherche Scientifique ( CNRS )
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STAT3 Transcription Factor ,0301 basic medicine ,Cancer Research ,Colorectal cancer ,Biopsy ,Mice, Nude ,colorectal cancer ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,Mouse model of colorectal and intestinal cancer ,Biology ,medicine.disease_cause ,Molecular oncology ,[ SDV.CAN ] Life Sciences [q-bio]/Cancer ,STAT3 ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Growth factor receptor ,Cell Line, Tumor ,Genetics ,medicine ,Animals ,Humans ,HSP110 Heat-Shock Proteins ,Intestinal Mucosa ,Phosphorylation ,Molecular Biology ,Cell Proliferation ,Microsatellite instability ,Cell cycle ,medicine.disease ,Molecular biology ,digestive system diseases ,3. Good health ,030104 developmental biology ,030220 oncology & carcinogenesis ,Cancer research ,biology.protein ,Female ,Colorectal Neoplasms ,Carcinogenesis ,Neoplasm Transplantation ,HSP110 ,Protein Binding - Abstract
IF 7.932; International audience; Heat shock protein 110 (HSP110) is induced by different stresses and, through its anti-apoptotic and chaperoning properties, helps cells survive these adverse situations. In colon cancers, HSP110 is abnormally abundant. We have recently shown that colorectal cancer patients with microsatellite instability (MSI) had an improved response to chemotherapy because they harbor an HSP110-inactivating mutation (HSP110DE9). In this work, we used patient biopsies, human colorectal cancer cells grown in vitro and in vivo (xenografts), and intestinal crypts to demonstrate that HSP110 is also involved in colon cancer growth. We showed that HSP110 induces colon cancer cell proliferation and that this effect is associated with STAT3 activation, specifically an increase in STAT3 phosphorylation, nuclear translocation and transcription factor activity. STAT3 inhibition blocks the proliferative effect of HSP110. From a molecular standpoint, we demonstrated that HSP110 directly binds to STAT3, thereby facilitating its phosphorylation by JAK2. Finally, we showed a correlation between HSP110 expression and STAT3 phosphorylation in colon cancer patient samples. Thus, the expression of HSP110 in colon cancer contributes to STAT3-dependent tumor growth and the frequent inactivating mutation of this chaperone is probably an important event underlying the improved prognosis in colon cancer displaying MSI.
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- 2017
5. Abstract LB-017: HSP110 sustains aberrant NFkB signaling in activated B-cell diffuse large B-cell lymphoma through MyD88 stabilization
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Laurent Martin, Oliver Olaf Wolz, Els Verhoeyen, Thierry Fest, Gaëtan Jego, Christophe Boudesco, Sebastien Causse, Arlette Hammann, Alexander N.R. Weber, Carmen Garrido, Université de Bourgogne ( UB ), Institut National de la Santé et de la Recherche Médicale ( INSERM ), CHU Dijon, Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand ( CHU Dijon ), and University of Tübingen
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Cancer Research ,Oncogene ,Biology ,medicine.disease ,Lymphoma ,[ SDV.CAN ] Life Sciences [q-bio]/Cancer ,Small hairpin RNA ,medicine.anatomical_structure ,Oncology ,Cell culture ,immune system diseases ,hemic and lymphatic diseases ,Cancer cell ,medicine ,Cancer research ,Gene silencing ,Diffuse large B-cell lymphoma ,neoplasms ,B cell - Abstract
Diffuse large B cell lymphoma (DLBCL) is an aggressive lymphoproliferative disorder of B lymphocytes accounting for 30 % of adult Non Hodgkin Lymphoma (NHL). Among DLBCL, Activated B Cell - DLBCL (ABC-DLBCL) is the most aggressive form and has a poor prognosis. Heat-shock proteins (HSPs) are molecular chaperons highly expressed in cancer cells and implicated in resistance to radio- and chemotherapy. Therefore, HSPs are envisioned as therapeutic targets in many cancers. Among the different HSPs, HSP110 has been recently identified as a pro-survival factor in germinal center-derived DLBCL (GC-DLBCL), through stabilization of the GC-DLBCL oncogene Bcl-6. Here, we have explored if HSP110 could also be involved in the survival of the most aggressive form of DLBCL. We observed a high HSP110 expression in all ABC-DLBCL patient samples, compared to normal reactive lymph nodes by using IHC staining of ABC-DLBCL tumor sections and transcriptional analysis of ABC-DLBCL patient tumors. Furthermore, shRNA silencing of HSP110 decreases the survival of several ABC-DLBCL cell lines, and downregulates the expression of pro-survival factors such as BcL2 and BcL-XL. SiRNA silencing of HSP110 abrogates NF-kB signaling, which is the major oncogenic pathway in ABC-DLBCL cell lines. In accord with these results, over-expression of HSP110 in DLBCL and non-DLBCL cell lines increases NF-kB signaling, indicating a tight interplay between HSP110 and the NF-kB pathway. Using immune-precipitation and DuolinkTM assays, we identified an in vitro and in cellulo interaction between HSP110 and Myd88, a critical protein of the NF-kB pathway that bears an activated mutation in many ABC-DLBC patients and that is responsible for lymphoma aggressiveness. Finally, we demonstrate that HSP110 stabilizes the wild type as well as the mutated form of Myd88, therefore facilitating the chronic NF-kB pathway activation in those cells. In conclusion, we identified HSP110 as a regulator of NF-kB signaling through MyD88 stabilization in ABC-DLBCL. This finding highlights HSP110 as a new potential therapeutic target in DLBCL. Citation Format: Christophe Boudesco, Sebastien Causse, Arlette Hammann, Els Verhoeyen, Laurent Martin, Thierry Fest, Oliver Wolz, Alexander Weber, Carmen Garrido, Gaetan Jego. HSP110 sustains aberrant NFkB signaling in activated B-cell diffuse large B-cell lymphoma through MyD88 stabilization [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-017. doi:10.1158/1538-7445.AM2017-LB-017
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- 2017
6. Transfer of functional microRNAs between glioblastoma and microvascular endothelial cells through gap junctions
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Eric Solary, Jonathan Boucher, Nicolas Pernet, Carmen Garrido, Dominique Thuringer, Gaëtan Jego, Laurent Cronier, Arlette Hammann, Lipides - Nutrition - Cancer (U866) (LNC), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA), Signalisation et Transports Ioniques Membranaires (STIM), Université de Poitiers-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS), Hématopoïèse normale et pathologique (U1170 Inserm), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR), Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] (UNICANCER/CRLCC-CGFL), UNICANCER, Université de Poitiers-Université de Tours-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Lipides - Nutrition - Cancer (U866) ( LNC ), Université de Bourgogne ( UB ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon ( ENSBANA ), Signalisation et Transports Ioniques Membranaires ( STIM ), Université de Poitiers-Centre National de la Recherche Scientifique ( CNRS ), Laboratoire Chrono-environnement ( LCE ), Université Bourgogne Franche-Comté ( UBFC ) -Centre National de la Recherche Scientifique ( CNRS ) -Université de Franche-Comté ( UFC ), Hématopoïèse normale et pathologique ( U1170 Inserm ), Université Paris-Sud - Paris 11 ( UP11 ) -Institut Gustave Roussy ( IGR ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), and Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc ( CRLCC - CGFL )
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0301 basic medicine ,Survival ,Angiogenesis ,Gene Expression ,Connexin ,Expression ,Cell Communication ,RNA Transport ,Microrna ,[ SDV.CAN ] Life Sciences [q-bio]/Cancer ,Invasion ,U87 ,Cancer ,Tumor ,Neovascularization, Pathologic ,Progression ,Gap junction ,Gap Junctions ,Brain ,Tubulogenesis ,Cell biology ,medicine.anatomical_structure ,Oncology ,Gap Junction ,Research Paper ,medicine.drug ,Cell type ,Endothelium ,Carbenoxolone ,Glioma-Cells ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,United-States ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,Biology ,03 medical and health sciences ,Cell Line, Tumor ,microRNA ,medicine ,Humans ,neoplasms ,[ SDV.BC ] Life Sciences [q-bio]/Cellular Biology ,Endothelial Cells ,nervous system diseases ,body regions ,MicroRNAs ,030104 developmental biology ,Intercellular Communication ,Immunology ,Glioblastoma - Abstract
IF 5.008; International audience; Extensive invasion and angiogenesis are hallmark features of malignant glioblastomas. Here, we co-cultured U87 human glioblastoma cells and human microvascular endothelial cells (HMEC) to demonstrate the exchange of microRNAs that initially involve the formation of gap junction communications between the two cell types. The functional inhibition of gap junctions by carbenoxolone blocks the transfer of the anti-tumor miR-145-5p from HMEC to U87, and the transfer of the pro-invasive miR-5096 from U87 to HMEC. These two microRNAs exert opposite effects on angiogenesis in vitro. MiR-5096 was observed to promote HMEC tubulogenesis, initially by increasing Cx43 expression and the formation of heterocellular gap junctions, and secondarily through a gap-junction independent pathway. Our results highlight the importance of microRNA exchanges between tumor and endothelial cells that in part involves the formation of functional gap junctions between the two cell types.
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- 2016
7. Plasma phospholipid transfer protein (PLTP) modulates adaptive immune functions through alternation of T helper cell polarization
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Stéphanie Lemaire-Ewing, Anabelle Sequeira-Le Grand, Valérie Deckert, Catherine Desrumaux, Nicolas Ogier, Naim Akhtar Khan, Jean-Paul Pais de Barros, Akadiri Yessoufou, Arlette Hammann, Julien Guy, Naig Le Guern, Laurent Lagrost, Laboratoire d'Excellence : Lipoprotéines et Santé : prévention et Traitement des maladies Inflammatoires et du Cancer ( LabEx LipSTIC ), Institut National de la Recherche Agronomique ( INRA ) -Université Montpellier 2 - Sciences et Techniques ( UM2 ) -Université Paris-Sud - Paris 11 ( UP11 ) -École pratique des hautes études ( EPHE ) -Institut Gustave Roussy ( IGR ) -Centre Hospitalier Régional Universitaire de Nancy ( CHRU Nancy ) -Université de Bourgogne ( UB ) -Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand ( CHU Dijon ) -Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc ( CRLCC - CGFL ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Fédération Francophone de la Cancérologie Digestive, FFCD-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-France-Comté] ( EFS [Bourgogne-France-Comté] ) -Centre National de la Recherche Scientifique ( CNRS ) -Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Université de Franche-Comté ( UFC ), Lipides - Nutrition - Cancer (U866) ( LNC ), Université de Bourgogne ( UB ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon ( ENSBANA ), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand ( CHU Dijon ), Plateforme cytométrie en flux [Fédération de Recherche Santé STIC de l’Université de Bourgogne], Université de Bourgogne ( UB ) -IFR100 - Structure fédérative de recherche Santé-STIC, Laboratoire d'Excellence : Lipoprotéines et Santé : prévention et Traitement des maladies Inflammatoires et du Cancer (LabEx LipSTIC), Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Paris-Sud - Paris 11 (UP11)-École pratique des hautes études (EPHE)-Institut Gustave Roussy (IGR)-Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy)-Université de Bourgogne (UB)-Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon)-Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc (CRLCC - CGFL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Fédération Francophone de la Cancérologie Digestive, FFCD-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-France-Comté] (EFS [Bourgogne-France-Comté])-Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire [Besançon] (CHRU Besançon)-Université de Franche-Comté (UFC)-Université de Montpellier (UM), Lipides - Nutrition - Cancer (U866) (LNC), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon), Université de Bourgogne (UB)-IFR100 - Structure fédérative de recherche Santé-STIC, Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Paris-Sud - Paris 11 (UP11)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Gustave Roussy (IGR)-Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy)-Université de Bourgogne (UB)-Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon)-Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] (UNICANCER/CRLCC-CGFL), UNICANCER-UNICANCER-Institut National de la Santé et de la Recherche Médicale (INSERM)-Fédération Francophone de la Cancérologie Digestive, FFCD-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université de Montpellier (UM), université de Bourgogne, LNC, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Gustave Roussy (IGR)-Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy)-Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Université de Bourgogne (UB)-Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon)-Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] (UNICANCER/CRLCC-CGFL), UNICANCER-UNICANCER-Institut National de la Santé et de la Recherche Médicale (INSERM)-Fédération Francophone de la Cancérologie Digestive, FFCD-Université de Montpellier (UM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), and Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)
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0301 basic medicine ,Lymphocyte ,Ipid Transfer Protein ,Adaptive Immunity ,Cardiovascular-Disease ,T-Lymphocytes, Regulatory ,Lipoprotein Metabolism ,Leukocyte Count ,Phospholipid transfer protein ,Polarization ,Immunology and Allergy ,[ SDV.IMM ] Life Sciences [q-bio]/Immunology ,Hypersensitivity, Delayed ,Phospholipid Transfer Proteins ,Cell Polarity ,Cell Differentiation ,T-Lymphocytes, Helper-Inducer ,T helper cell ,Flow Cytometry ,Acquired immune system ,Cell biology ,Infectious Diseases ,medicine.anatomical_structure ,Endothelial-Cells ,Cytokines ,[SDV.IMM]Life Sciences [q-bio]/Immunology ,medicine.symptom ,Research Article ,Density-Lipoprotein ,[SDV.IMM] Life Sciences [q-bio]/Immunology ,Human Atherosclerotic Plaques ,T cell ,Circulating Interleukin-18 ,Immunology ,T Cell ,Antigen-Presenting Cells ,Inflammation ,Acute Myocardial-Infarction ,GATA3 Transcription Factor ,Biology ,03 medical and health sciences ,Immune system ,medicine ,Animals ,Antigen-presenting cell ,Deficient Mice ,Alpha-Tocopherol ,Mice, Inbred C57BL ,030104 developmental biology ,Vitamin-E ,T-Box Domain Proteins ,Biomarkers ,Spleen - Abstract
International audience; Objective: Plasma phospholipid transfer protein (PLTP) is a key determinant of lipoprotein metabolism, and both animal and human studies converge to indicate that PLTP promotes atherogenesis and its thromboembolic complications. Moreover, it has recently been reported that PLTP modulates inflammation and immune responses. Although earlier studies from our group demonstrated that PLTP can modify macrophage activation, the implication of PLTP in the modulation of T-cell-mediated immune responses has never been investigated and was therefore addressed in the present study. Approach and results: In the present study, we demonstrated that PLTP deficiency in mice has a profound effect on CD4(+) Th0 cell polarization, with a shift towards the anti-inflammatory Th2 phenotype under both normal and pathological conditions. In a model of contact hypersensitivity, a significantly impaired response to skin sensitization with the hapten-2,4-dinitrofluorobenzene (DNFB) was observed in PLTP-deficient mice compared to wild-type (WT) mice. Interestingly, PLTP deficiency in mice exerted no effect on the counts of total white blood cells, lymphocytes, granulocytes, or monocytes in the peripheral blood. Moreover, PLTP deficiency did not modify the amounts of CD4(+) and CD8(+) T lymphocyte subsets. However, PLTP-deficiency, associated with upregulation of the Th2 phenotype, was accompanied by a significant decrease in the production of the pro-Th1 cytokine interleukin 18 by accessory cells. Conclusions: For the first time, this work reports a physiological role for PLTP in the polarization of CD4(+) T cells toward the pro-inflammatory Th1 phenotype.
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- 2016
8. Inhibition of HSP27 blocks fibrosis development and EMT features by promoting Snail degradation
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Martin E. Gleave, Pierre-Simon Bellaye, Joëlle Marchal-Somme, Carmen Garrido, Philippe Camus, Andreas Günther, Olivier Micheau, Martin Kolb, Guillaume Wettstein, Bruno Crestani, Adonis Hazoumé, Philippe Bonniaud, Arlette Hammann, J Gauldie, and Paul Soler
- Subjects
endocrine system ,Pathology ,medicine.medical_specialty ,Epithelial-Mesenchymal Transition ,animal structures ,Snails ,HSP27 Heat-Shock Proteins ,Biology ,Biochemistry ,Cell Line ,Rats, Sprague-Dawley ,Transforming Growth Factor beta1 ,03 medical and health sciences ,Idiopathic pulmonary fibrosis ,0302 clinical medicine ,In vivo ,Fibrosis ,Pulmonary fibrosis ,Genetics ,medicine ,Animals ,Humans ,Epithelial–mesenchymal transition ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,Gene knockdown ,Epithelial Cells ,Oligonucleotides, Antisense ,Thionucleotides ,Cadherins ,medicine.disease ,Rats ,3. Good health ,030220 oncology & carcinogenesis ,embryonic structures ,Cancer research ,Myofibroblast ,Transcription Factors ,Biotechnology ,Transforming growth factor - Abstract
Idiopathic pulmonary fibrosis (IPF) is a devastating disease characterized by myofibroblast proliferation. Transition of epithelial/mesothelial cells into myofibroblasts [epithelial-to-mesenchymal transition (EMT)] occurs under the influence of transforming growth factor (TGF)-β1, with Snail being a major transcription factor. We study here the role of the heat-shock protein HSP27 in fibrogenesis and EMT. In vitro, we have up- and down-modulated HSP27 expression in mesothelial and epithelial cell lines and studied the expression of different EMT markers induced by TGF-β1. In vivo, we inhibited HSP27 with the antisense oligonucleotide OGX-427 (in phase II clinical trials as anticancer agent) in our rat subpleural/pulmonary fibrosis models. We demonstrate that HSP27 is strongly expressed during the fibrotic process in patients with IPF and in different in vivo models. We showed that HSP27 binds to and stabilizes Snail and consequently induces EMT. Conversely, HSP27 knockdown leads to Snail proteasomal degradation, thus inhibiting TGF-β1-induced EMT. Inhibition of HSP27 with OGX-427 efficiently blocks EMT and fibrosis development. Controls in vivo were an empty adenovirus that did not induce fibrosis and a control antisense oligonucleotide. The present work opens the possibility of a new therapeutic use for HSP27 inhibitors against IPF, for which there is no conclusively effective treatment.
- Published
- 2013
9. The HSP90 inhibitor, 17AAG, protects the intestinal stem cell niche and inhibits graft versus host disease development
- Author
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Arlette Hammann, Eric Solary, Oleg N. Demidov, Gaëtan Jego, Robinet E, A. Seignez, Sarah Richaud, Anastasia R. Goloudina, Evelyne Kohli, Magali Svrcek, Carmen Garrido, Eric Fourmaux, Deepti A, Anne-Laure Joly, Sophie Hébrard, Ada Collura, Elise Schmitt, Lipides - Nutrition - Cancer (U866) (LNC), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA), Laboratoire d'Excellence : Lipoprotéines et Santé : prévention et Traitement des maladies Inflammatoires et du Cancer (LabEx LipSTIC), Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Paris-Sud - Paris 11 (UP11)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Gustave Roussy (IGR)-Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy)-Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Université de Bourgogne (UB)-Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon)-Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] (UNICANCER/CRLCC-CGFL), UNICANCER-UNICANCER-Institut National de la Santé et de la Recherche Médicale (INSERM)-Fédération Francophone de la Cancérologie Digestive, FFCD-Université de Montpellier (UM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS BFC)-Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), CHU Dijon, Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon), Centre de Recherche Saint-Antoine (UMRS893), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Interaction virus-hôte et maladies du foie, Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Hématopoïèse normale et pathologique, Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] (UNICANCER/CRLCC-CGFL), UNICANCER, Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Paris-Sud - Paris 11 (UP11)-École pratique des hautes études (EPHE)-Institut Gustave Roussy (IGR)-Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy)-Université de Bourgogne (UB)-Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon)-Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc (CRLCC - CGFL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Fédération Francophone de la Cancérologie Digestive, FFCD-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-France-Comté] (EFS [Bourgogne-France-Comté])-Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire [Besançon] (CHRU Besançon)-Université de Franche-Comté (UFC)-Université de Montpellier (UM), Centre de Recherche Saint-Antoine (CR Saint-Antoine), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Gustave Roussy (IGR)-Université Paris-Sud - Paris 11 (UP11), Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc (CRLCC - CGFL), Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Paris-Sud - Paris 11 (UP11)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Gustave Roussy (IGR)-Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy)-Université de Bourgogne (UB)-Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon)-Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] (UNICANCER/CRLCC-CGFL), UNICANCER-UNICANCER-Institut National de la Santé et de la Recherche Médicale (INSERM)-Fédération Francophone de la Cancérologie Digestive, FFCD-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université de Montpellier (UM), Lipides - Nutrition - Cancer (U866) ( LNC ), Université de Bourgogne ( UB ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon ( ENSBANA ), Laboratoire d'Excellence : Lipoprotéines et Santé : prévention et Traitement des maladies Inflammatoires et du Cancer ( LabEx LipSTIC ), Institut National de la Recherche Agronomique ( INRA ) -Université Montpellier 2 - Sciences et Techniques ( UM2 ) -Université Paris-Sud - Paris 11 ( UP11 ) -École pratique des hautes études ( EPHE ) -Institut Gustave Roussy ( IGR ) -Centre Hospitalier Régional Universitaire de Nancy ( CHRU Nancy ) -Université de Bourgogne ( UB ) -Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand ( CHU Dijon ) -Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc ( CRLCC - CGFL ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Fédération Francophone de la Cancérologie Digestive, FFCD-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-France-Comté] ( EFS [Bourgogne-France-Comté] ) -Centre National de la Recherche Scientifique ( CNRS ) -Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Université de Franche-Comté ( UFC ), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand ( CHU Dijon ), Centre de Recherche Saint-Antoine ( CR Saint-Antoine ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université Paris-Sud - Paris 11 ( UP11 ) -Institut Gustave Roussy ( IGR ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc ( CRLCC - CGFL ), and UNICANCER-UNICANCER-Institut National de la Santé et de la Recherche Médicale (INSERM)-Fédération Francophone de la Cancérologie Digestive, FFCD-Université de Montpellier (UM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC)
- Subjects
0301 basic medicine ,X-Box Binding Protein 1 ,Cancer Research ,Lactams, Macrocyclic ,RNA Splicing ,T-Cells ,Graft vs Host Disease ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,Biology ,[ SDV.CAN ] Life Sciences [q-bio]/Cancer ,Hsp90 inhibitor ,03 medical and health sciences ,Mice ,Sensitivity ,Inflammatory-Bowel-disease ,Genetics ,medicine ,Benzoquinones ,Animals ,[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology ,Neural progenitor cells ,HSP90 Heat-Shock Proteins ,Intestinal Mucosa ,Stem Cell Niche ,[ SDV.GEN.GH ] Life Sciences [q-bio]/Genetics/Human genetics ,[ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology ,Molecular Biology ,Leukemia ,[ SDV.BC ] Life Sciences [q-bio]/Cellular Biology ,Bone-Marrow-Transplantation ,Molecules ,medicine.disease ,Stem cell niche ,3. Good health ,Ire1-Alpha ,Intestines ,Mice, Inbred C57BL ,030104 developmental biology ,Graft-versus-host disease ,Er Stress ,[SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics ,Cytoprotection ,Immunology ,Multiple-Myeloma ,Female - Abstract
IF 7.932; International audience; Graft versus host disease (GvHD), which is the primary complication of allogeneic bone marrow transplantation, can alter the intestinal barrier targeted by activated donor T-cells. Chemical inhibition of the stress protein HSP90 was demonstrated in vitro to inhibit T-cell activation and to modulate endoplasmic reticulum (ER) stress to which intestinal cells are highly susceptible. Since the HSP90 inhibitor 17-allylamino-demethoxygeldanamycin (17AAG) is developed in clinics, we explored here its ability to control intestinal acute GvHD in vivo in two mouse GvHD models (C57BL/6 -> BALB/c and FVB/N -> Lgr5-eGFP), ex vivo in intestine organoids and in vitro in intestinal epithelial cultures. We show that 17AAG decreases GvHD-associated mortality without impairing graft versus leukemia effect. While 17AAG effect in T-cell activation is just moderate at the dose used in vivo, we observe a striking intestinal integrity protection. At the intestine level, the drug promotes the splicing of the transcription factor X-box binding protein 1 (XBP1), which is a key component of the ER stress. This effect is associated with a decrease in intestinal damage and an increase in Lgr5(+) stem cells, Paneth cells and defensins production. The importance of XBP1 splicing control is further confirmed in cultured cells and organoids of primary intestinal epithelium where XBP1 is either shRNA depleted or inhibited with toyocamycin. In conclusion, 17AAG has a protective effect on the epithelial intestinal barrier in mouse models of acute GvHD. This compound deserves to be tested in the therapeutic control of acute GvHD.
- Published
- 2016
10. Restoring Anticancer Immune Response by Targeting Tumor-Derived Exosomes With a HSP70 Peptide Aptamer
- Author
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Cédric Rébé, Hajare Mjahed, Nicolas Pernet, Victor Clausse, Carmen Garrido, Renaud Seigneuric, Sarah Richaud, Pierre Fumoleau, Alexandre M.M. Dias, François Ghiringhelli, Guillaume Marcion, Nicolas Isambert, Vincent Goussot, Marine Cordonnier, Frédéric Lirussi, Jessica Gobbo, Aurélie de Thonel, Aurélie Bertaut, Arlette Hammann, Lipides - Nutrition - Cancer (U866) (LNC), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA), Laboratoire d'Excellence : Lipoprotéines et Santé : prévention et Traitement des maladies Inflammatoires et du Cancer (LabEx LipSTIC), Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Paris-Sud - Paris 11 (UP11)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Gustave Roussy (IGR)-Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy)-Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Université de Bourgogne (UB)-Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon)-Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] (UNICANCER/CRLCC-CGFL), UNICANCER-UNICANCER-Institut National de la Santé et de la Recherche Médicale (INSERM)-Fédération Francophone de la Cancérologie Digestive, FFCD-Université de Montpellier (UM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS BFC)-Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Département d'oncologie médicale [Centre Georges-François Leclerc], Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] (UNICANCER/CRLCC-CGFL), UNICANCER-UNICANCER, Institut Charles Sadron (ICS), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Matériaux et Nanosciences Grand-Est (MNGE), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-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 de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), UNICANCER, Unité de biostatistiques [Centre Georges-François Leclerc], Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon), Physiologie de la Nutrition et Toxicologie (NUTox) (U866, Lipides et nutrition, équipe 7) (NUTox), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA)-Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA), Institut National du Cancer, Conseil Regional de Bourgogne, ANR-11-LABX-0021,Lipstic,Lipoprotéines et santé : prévention et traitement des maladies inflammatoires non vasculaires et du cancer(2011), European Project: 248835,EC:FP7:ICT,FP7-ICT-2009-4,SPEDOC(2010), Centre épigénétique et destin cellulaire (EDC (UMR_7216)), Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Paris-Sud - Paris 11 (UP11)-École pratique des hautes études (EPHE)-Institut Gustave Roussy (IGR)-Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy)-Université de Bourgogne (UB)-Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon)-Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc (CRLCC - CGFL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Fédération Francophone de la Cancérologie Digestive, FFCD-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-France-Comté] (EFS [Bourgogne-France-Comté])-Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire [Besançon] (CHRU Besançon)-Université de Franche-Comté (UFC)-Université de Montpellier (UM), Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc (CRLCC - CGFL), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Matériaux et nanosciences d'Alsace, Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), ANR-11-LABX-0021/11-LABX-0021,Lipstic,Lipoprotéines et santé : prévention et traitement des maladies inflammatoires non vasculaires et du(2011), Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Paris-Sud - Paris 11 (UP11)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Gustave Roussy (IGR)-Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy)-Université de Bourgogne (UB)-Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon)-Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] (UNICANCER/CRLCC-CGFL), UNICANCER-UNICANCER-Institut National de la Santé et de la Recherche Médicale (INSERM)-Fédération Francophone de la Cancérologie Digestive, FFCD-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université de Montpellier (UM), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), ANR-11-LABX-0021,Lipstic,Lipoprotéines et santé : prévention et traitement des maladies inflammatoires non vasculaires et du(2011), UNICANCER-UNICANCER-Institut National de la Santé et de la Recherche Médicale (INSERM)-Fédération Francophone de la Cancérologie Digestive, FFCD-Université de Montpellier (UM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), and Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Matériaux et nanosciences d'Alsace (FMNGE)
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Male ,0301 basic medicine ,Cancer Research ,Lung Neoplasms ,[SDV]Life Sciences [q-bio] ,Exosomes ,Mice ,Mechanisms ,Myeloid Cells ,ComputingMilieux_MISCELLANEOUS ,Cancer ,Ovarian Neoplasms ,3. Good health ,Phenotype ,Oncology ,Colonic Neoplasms ,Female ,medicine.drug ,Myeloid suppressor-Cells ,B-cells ,Antineoplastic Agents ,Breast Neoplasms ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,Biology ,03 medical and health sciences ,Lymphocytes, Tumor-Infiltrating ,Immune system ,Cell Line, Tumor ,medicine ,Animals ,Humans ,HSP70 Heat-Shock Proteins ,Cell Proliferation ,Cisplatin ,Macrophages ,Neoplasms, Experimental ,medicine.disease ,Molecular biology ,Toll-Like Receptor 2 ,Microvesicles ,Mice, Inbred C57BL ,Membrane heat-shock-protein-70 Hsp70 ,TLR2 ,Interferometry ,030104 developmental biology ,Tumor progression ,Release ,Cancer cell ,Myeloid-derived Suppressor Cell ,Aptamers, Peptide ,Spleen - Abstract
International audience; Background: Exosomes, via heat shock protein 70 (HSP70) expressed in their membrane, are able to interact with the toll-like receptor 2 (TLR2) on myeloid-derived suppressive cells (MDSCs), thereby activating them.Methods: We analyzed exosomes from mouse (C57Bl/6) and breast, lung, and ovarian cancer patient samples and cultured cancer cells with different approaches, including nanoparticle tracking analysis, biolayer interferometry, FACS, and electron microscopy. Data were analyzed with the Student's t and Mann-Whitney tests. All statistical tests were two-sided.Results: We showed that the A8 peptide aptamer binds to the extracellular domain of membrane HSP70 and used the aptamer to capture HSP70 exosomes from cancer patient samples. The number of HSP70 exosomes was higher in cancer patients than in healthy donors (mean, ng/mL +/- SD = 3.5 +/- 1.7 vs 0.17 +/- 0.11, respectively, P = .004). Accordingly, all cancer cell lines examined abundantly released HSP70 exosomes, whereas "normal" cells did not. HSP70 had higher affinity for A8 than for TLR2; thus, A8 blocked HSP70/TLR2 association and the ability of tumor-derived exosomes to activate MDSCs. Treatment of tumor-bearing C57Bl/6 mice with A8 induced a decrease in the number of MDSCs in the spleen and inhibited tumor progression (n = 6 mice per group). Chemotherapeutic agents such as cisplatin or 5FU increase the amount of HSP70 exosomes, favoring the activation of MDSCs and hampering the development of an antitumor immune response. In contrast, this MDSC activation was not observed if cisplatin or 5FU was combined with A8. As a result, the antitumor effect of the drugs was strongly potentiated.Conclusions: A8 might be useful for quantifying tumor-derived exosomes and for cancer therapy through MDSC inhibition.
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- 2016
11. Extracellular HSP110 skews macrophage polarization in colorectal cancer
- Author
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Gaëtan Jego, Carmen Garrido, Sebastien Causse, Nadhir Yousfi, Magali Svrcek, Alex Duval, Christophe Boudesco, Kevin Berthenet, Arlette Hammann, Laurence Duplomb, Kristell Wanherdrick, Sarah Richaud, Ada Collura, Laboratoire d'Excellence : Lipoprotéines et Santé : prévention et Traitement des maladies Inflammatoires et du Cancer ( LabEx LipSTIC ), Institut National de la Recherche Agronomique ( INRA ) -Université Montpellier 2 - Sciences et Techniques ( UM2 ) -Université Paris-Sud - Paris 11 ( UP11 ) -École pratique des hautes études ( EPHE ) -Institut Gustave Roussy ( IGR ) -Centre Hospitalier Régional Universitaire de Nancy ( CHRU Nancy ) -Université de Bourgogne ( UB ) -Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand ( CHU Dijon ) -Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc ( CRLCC - CGFL ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Fédération Francophone de la Cancérologie Digestive, FFCD-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-France-Comté] ( EFS [Bourgogne-France-Comté] ) -Centre National de la Recherche Scientifique ( CNRS ) -Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Université de Franche-Comté ( UFC ), Lipides - Nutrition - Cancer (U866) ( LNC ), Université de Bourgogne ( UB ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon ( ENSBANA ), Centre de Recherche Saint-Antoine ( CR Saint-Antoine ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Laboratoire d'Immunologie et Immunothérapie des Cancers ( LIIC ), Université de Bourgogne ( UB ) -École pratique des hautes études ( EPHE ), Centre de génétique - Centre de référence des maladies rares, anomalies du développement et syndromes malformatifs (CHU de Dijon), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand ( CHU Dijon ), Génétique des Anomalies du Développement ( GAD ), Université de Bourgogne ( UB ) -IFR100 - Structure fédérative de recherche Santé-STIC, Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc ( CRLCC - CGFL ), Laboratoire d'Excellence : Lipoprotéines et Santé : prévention et Traitement des maladies Inflammatoires et du Cancer (LabEx LipSTIC), Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Paris-Sud - Paris 11 (UP11)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Gustave Roussy (IGR)-Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy)-Université de Bourgogne (UB)-Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon)-Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] (UNICANCER/CRLCC-CGFL), UNICANCER-UNICANCER-Institut National de la Santé et de la Recherche Médicale (INSERM)-Fédération Francophone de la Cancérologie Digestive, FFCD-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université de Montpellier (UM), Lipides - Nutrition - Cancer (U866) (LNC), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA), Centre de Recherche Saint-Antoine (UMRS893), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire d'Immunologie et Immunothérapie des Cancers (LIIC), Université de Bourgogne (UB)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon), Génétique des Anomalies du Développement (GAD), IFR100 - Structure fédérative de recherche Santé-STIC-Université de Bourgogne (UB), Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] (UNICANCER/CRLCC-CGFL), UNICANCER, Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Paris-Sud - Paris 11 (UP11)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Gustave Roussy (IGR)-Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy)-Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Université de Bourgogne (UB)-Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon)-Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] (UNICANCER/CRLCC-CGFL), UNICANCER-UNICANCER-Institut National de la Santé et de la Recherche Médicale (INSERM)-Fédération Francophone de la Cancérologie Digestive, FFCD-Université de Montpellier (UM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS BFC)-Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université de Bourgogne (UB), Université de Bourgogne (UB)-IFR100 - Structure fédérative de recherche Santé-STIC, UNICANCER-UNICANCER-Institut National de la Santé et de la Recherche Médicale (INSERM)-Fédération Francophone de la Cancérologie Digestive, FFCD-Université de Montpellier (UM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), École pratique des hautes études (EPHE), Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Paris-Sud - Paris 11 (UP11)-École pratique des hautes études (EPHE)-Institut Gustave Roussy (IGR)-Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy)-Université de Bourgogne (UB)-Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon)-Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc (CRLCC - CGFL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Fédération Francophone de la Cancérologie Digestive, FFCD-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-France-Comté] (EFS [Bourgogne-France-Comté])-Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire [Besançon] (CHRU Besançon)-Université de Franche-Comté (UFC)-Université de Montpellier (UM), Centre de Recherche Saint-Antoine (CR Saint-Antoine), École pratique des hautes études (EPHE)-Université de Bourgogne (UB), and Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc (CRLCC - CGFL)
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0301 basic medicine ,Macrophage ,Immunology ,Macrophage polarization ,Activation ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,Expression ,Biology ,Colon-Cancer ,[ SDV.CAN ] Life Sciences [q-bio]/Cancer ,03 medical and health sciences ,0302 clinical medicine ,In vivo ,Polarization ,Extracellular ,medicine ,Immunology and Allergy ,Cytotoxic T cell ,Kappa-B ,Colorectal ,Heat-Shock Proteins ,Original Research ,Cancer ,Tumor-infiltrating lymphocytes ,Heat-Shock Protein ,Dendritic Cells ,medicine.disease ,Ikk-Beta ,Molecular biology ,3. Good health ,030104 developmental biology ,Oncology ,030220 oncology & carcinogenesis ,TLR4 ,Tumor-Infiltrating Lymphocytes ,Microsatellite Instability ,Chaperone Machines - Abstract
IF 7.644; International audience; HSP110 is induced by different stresses and, through its anti-apoptotic and chaperoning properties, helps the cells to survive these adverse situations. In colon cancers, HSP110 is abnormally abundant. We have recently showed that colorectal cancer (CRC) patients with microsatellite instability (MSI) had an improved response to chemotherapy because they harbor an HSP110 inactivating mutation (HSP110DE9). In this work, we have used patients' biopsies and human CRC cells grown in vitro and in vivo (xenografts) to demonstrate that (1) HSP110 is secreted by CRC cells and that the amount of this extracellular HSP110 is strongly decreased by the expression of the mutant HSP110DE9, (2) Supernatants from CRC cells overexpressing HSP110 or purified recombinant human HSP110 (LPS-free) affect macrophage differentiation/polarization by favoring a pro-tumor, anti-inflammatory profile, (3) Conversely, inhibition of HSP110 (expression of siRNA, HSP110DE9 or immunodepletion) induced the formation of macrophages with a cytotoxic, pro-inflammatory profile. (4) Finally, this effect of extracellular HSP110 on macrophages seems to implicate TLR4. These results together with the fact that colorectal tumor biopsies with HSP110 high were infiltrated with macrophages with a pro-tumoral profile while those with HSP110 low were infiltrated with macrophages with a cytotoxic profile, suggest that the effect of extracellular HSP110 function on macrophages may also contribute to the poor outcomes associated with HSP110 expression.
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- 2016
12. α-Tocopherol Modulates Phosphatidylserine Externalization in Erythrocytes
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Naig Le Guern, Xian-Cheng Jiang, Anne Athias, Arlette Hammann, David Masson, Martina Schneider, Laurent Lagrost, Catherine Desrumaux, Jean-Paul Pais de Barros, Valérie Deckert, Fabienne Dutrillaux, and Alexis Klein
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Vitamin ,medicine.medical_specialty ,Erythrocytes ,Whole Blood Coagulation Time ,medicine.medical_treatment ,alpha-Tocopherol ,Phospholipid ,Cell Separation ,Phosphatidylserines ,Biology ,Fibrin Fibrinogen Degradation Products ,Mice ,chemistry.chemical_compound ,Annexin ,In vivo ,Phospholipid transfer protein ,Internal medicine ,medicine ,Animals ,Tocopherol ,Phospholipid Transfer Proteins ,Blood Coagulation ,Mice, Knockout ,Vitamin E ,Erythrocyte Membrane ,Homozygote ,Phosphatidylserine ,Phenotype ,Endocrinology ,chemistry ,Biochemistry ,Cardiology and Cardiovascular Medicine ,Oxidation-Reduction ,Biomarkers - Abstract
Objective— The aim of the present study was to assess the effect of α-tocopherol, the main vitamin E isomer on phosphatidylserine (PS) exposure at the surface of circulating erythrocytes, and to determine consequences on erythrocyte properties. Methods and Results— In vitro α-tocopherol enrichment of isolated erythrocytes significantly decreased PS externalization as assessed by lower Annexin V-fluorescein isothiocyanate labeling. Plasma phospholipid transfer protein (PLTP) transfers vitamin E, and both α-and γ-tocopherol accumulated in circulating erythrocytes from PLTP-deficient homozygous (PLTP −/− ) mice as compared with wild-type mice. In agreement with in vitro studies, vitamin E–enriched erythrocytes from PLTP −/− mice displayed fewer externalized PS molecules than wild-type controls (−64%, P −/− erythrocytes was accompanied by impairment of their procoagulant properties, with a 20% increase in clotting time as compared with wild-type controls ( P −/− mice compared with wild-type mice. Conclusions— Vitamin E modifies PS externalization in circulating erythrocytes, thus modulating in vivo their PS-dependent procoagulant properties.
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- 2006
13. Heat Shock Protein 70 Neutralization Exerts Potent Antitumor Effects in Animal Models of Colon Cancer and Melanoma
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Loic Maingret, Guido Kroemer, François Ghiringhelli, Elise Schmitt, Anne-Laure Rérole, Eric Solary, Carmen Garrido, Pierre-Emmanuel Puig, and Arlette Hammann
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Cancer Research ,Programmed cell death ,Lactams, Macrocyclic ,Melanoma, Experimental ,CD8-Positive T-Lymphocytes ,Transfection ,Mice ,Rats, Nude ,Lymphocytes, Tumor-Infiltrating ,Heat shock protein ,Benzoquinones ,medicine ,Animals ,Humans ,Cytotoxic T cell ,HSP70 Heat-Shock Proteins ,biology ,Apoptosis Inducing Factor ,Cancer ,Drug Synergism ,medicine.disease ,Combined Modality Therapy ,Hsp90 ,Peptide Fragments ,Rats ,Hsp70 ,Mice, Inbred C57BL ,Disease Models, Animal ,Rifabutin ,Oncology ,Apoptosis ,Immunology ,Cancer cell ,biology.protein ,Cancer research ,Cisplatin ,Colorectal Neoplasms - Abstract
When overexpressed, the stress protein heat shock protein 70 (HSP70) increases the oncogenic potential of cancer cells in rodent models. HSP70 also prevents apoptosis, thereby increasing the survival of cells exposed to a wide range of otherwise lethal stimuli. These protective functions of HSP70 involve its interaction with and neutralization of the adaptor molecule apoptotic protease activation factor-1, implicated in caspase activation, and the flavoprotein apoptosis-inducing factor (AIF), involved in caspase-independent cell death. We have shown previously that a peptide containing the AIF sequence involved in its interaction with HSP70 (ADD70, amino acids 150-228) binds to and neutralizes HSP70 in the cytosol, thereby sensitizing cancer cells to apoptosis induced by a variety of death stimuli. Here, we show that expression of ADD70 in tumor cells decreases their tumorigenicity in syngeneic animals without affecting their growth in immunodeficient animals. ADD70 antitumorigenic effects are associated with an increase in tumor-infiltrating cytotoxic CD8+ T cells. In addition, ADD70 sensitizes rat colon cancer cells (PROb) and mouse melanoma cells (B16F10) to the chemotherapeutic agent cisplatin. ADD70 also shows an additive effect with HSP90 inhibition by 17-allylamino-17-demethoxygeldanamycin in vitro. Altogether, these data indicate the potential interest of targeting the HSP70 interaction with AIF for cancer therapy. (Cancer Res 2006; 66(8): 4191-7)
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- 2006
14. Chemotherapy enhances TNF-related apoptosis-inducing ligand DISC assembly in HT29 human colon cancer cells
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Eric Solary, Jürg Tschopp, Véronique Drouineaud, Olivier Micheau, Marie-Thérèse Dimanche-Boitrel, Arlette Hammann, and Sandrine Lacour
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Cancer Research ,Programmed cell death ,Antineoplastic Agents ,Apoptosis ,Ligands ,Caspase 8 ,Membrane Potentials ,TNF-Related Apoptosis-Inducing Ligand ,Genetics ,Humans ,Cytotoxic T cell ,FADD ,Molecular Biology ,Caspase ,Death domain ,Membrane Glycoproteins ,biology ,Tumor Necrosis Factor-alpha ,Caspase 9 ,Mitochondria ,Enzyme Activation ,Caspases ,Immunology ,Cancer cell ,Cancer research ,biology.protein ,Apoptosis Regulatory Proteins ,Carrier Proteins ,HT29 Cells ,BH3 Interacting Domain Death Agonist Protein - Abstract
Cytokines such as Fas-ligand (Fas-L) and Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL) can induce human colon cancer cell apoptosis through engagement of their death domain receptors. All the cancer cells are not sensitive to these cytokines. We have shown recently that low doses of cytotoxic drugs could restore TRAIL-induced cell death in resistant colon cancer cell lines. The present work further explores the death pathway triggered by the cytotoxic drug/TRAIL combination in HT-29 colon cancer cells (www.alexis-corp.com). Clinically relevant concentrations of cisplatin, doxorubicin and 5-fluorouracil synergize with TRAIL to trigger HT-29 cell death. Activation of this pathway leads to apoptosis that involves both caspases and the mitochondria. An increased recruitment of Fas-associated death domain (FADD) and procaspase-8 to the TRAIL-induced death-inducing signaling complex (DISC) was shown in cells exposed to anticancer drugs. Following caspase-8 activation at the DISC level, the mitochondria-dependent death pathway is activated, as demonstrated by the cleavage of Bid, the dissipation of DeltaPsi(m), the release of mitochondrial proteins in the cytosol and the inhibitory effect of Bcl-2 expression. Importantly, besides mitochondrial potentiation, we show here that cytotoxic drugs sensitize HT-29 colon cancer cells to TRAIL-induced cell death by enhancing FADD and procaspase-8 recruitment to the DISC, a novel mechanism whose efficacy could depend partly on Bcl-2 expression level.
- Published
- 2003
15. Tif1γ regulates the TGF-β1 receptor and promotes physiological aging of hematopoietic stem cells
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Laetitia Saint-Paul, Romain Z. Martin, Anne Largeot, Arlette Hammann, Ronan Quéré, Marie-Lorraine Chretien, Jean-Noël Bastie, Virginie Carmignac, Laurent Delva, and Jean-Paul Pais de Barros
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Aging ,Myeloid ,Receptor, Transforming Growth Factor-beta Type I ,Receptors, Cell Surface ,Cell Separation ,Biology ,Protein Serine-Threonine Kinases ,Transforming Growth Factor beta1 ,Mice ,Signaling Lymphocytic Activation Molecule Family Member 1 ,Antigens, CD ,medicine ,Animals ,Myeloid Cells ,RNA, Messenger ,Polyubiquitin ,Transcription factor ,Cellular Senescence ,Regulation of gene expression ,Multidisciplinary ,Ubiquitination ,hemic and immune systems ,Biological Sciences ,Hematopoietic Stem Cells ,Cell biology ,Hematopoiesis ,Haematopoiesis ,medicine.anatomical_structure ,Physiological Aging ,Phenotype ,Gene Expression Regulation ,Signal transduction ,Stem cell ,Cell aging ,Receptors, Transforming Growth Factor beta ,Signal Transduction ,Transcription Factors - Abstract
The hematopoietic system declines with age. Myeloid-biased differentiation and increased incidence of myeloid malignancies feature aging of hematopoietic stem cells (HSCs), but the mechanisms involved remain uncertain. Here, we report that 4-mo-old mice deleted for transcription intermediary factor 1γ (Tif1γ) in HSCs developed an accelerated aging phenotype. To reinforce this result, we also show that Tif1γ is down-regulated in HSCs during aging in 20-mo-old wild-type mice. We established that Tif1γ controls TGF-β1 receptor (Tgfbr1) turnover. Compared with young HSCs, Tif1γ(-/-) and old HSCs are more sensitive to TGF-β signaling. Importantly, we identified two populations of HSCs specifically discriminated by Tgfbr1 expression level and provided evidence of the capture of myeloid-biased (Tgfbr1(hi)) and myeloid-lymphoid-balanced (Tgfbr1(lo)) HSCs. In conclusion, our data provide a new paradigm for Tif1γ in regulating the balance between lymphoid- and myeloid-derived HSCs through TGF-β signaling, leading to HSC aging.
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- 2014
16. Regulation of the proapoptotic functions of prostate apoptosis response-4 (Par-4) by casein kinase 2 in prostate cancer cells
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Gaëtan Jego, Olivier Micheau, Adonis Hazoumé, Odile Filhol, Valérie Mezger, John E. Eriksson, Vivek M. Rangnekar, E Fourmaux, Vitaly Kochin, H Mjahed, Kimmo O. Isoniemi, Carmen Garrido, A de Thonel, Arlette Hammann, Palma Rocchi, Centre épigénétique et destin cellulaire (EDC (UMR_7216)), Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Lipides - Nutrition - Cancer (U866) (LNC), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA), Sapporo Medical University, Åbo Akademi University [Turku], Biologie du Cancer et de l'Infection (BCI ), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), 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)-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), Stress Cellulaire, Université de la Méditerranée - Aix-Marseille 2-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre épigénétique et destin cellulaire (EDC), Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), University of Kentucky (UK), Valérie, MEZGER, Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut National de la Santé et de la Recherche Médicale (INSERM), and University of Kentucky
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Male ,Cancer Research ,animal structures ,CK2 ,[SDV]Life Sciences [q-bio] ,Immunology ,Amino Acid Motifs ,PAWR ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,[SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC] ,Biology ,[SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology ,[SDV.MHEP.UN]Life Sciences [q-bio]/Human health and pathology/Urology and Nephrology ,Cellular and Molecular Neuroscience ,Prostate cancer ,[SDV.CAN] Life Sciences [q-bio]/Cancer ,Prostate ,Cell Line, Tumor ,[SDV.BC.BC] Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC] ,medicine ,Animals ,Humans ,Casein Kinase II ,ComputingMilieux_MISCELLANEOUS ,Gene knockdown ,Kinase ,phosphorylation ,fungi ,ta1182 ,apoptosis ,Prostatic Neoplasms ,[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology ,Cell Biology ,medicine.disease ,prostate cancer ,[SDV.MHEP.UN] Life Sciences [q-bio]/Human health and pathology/Urology and Nephrology ,Rats ,medicine.anatomical_structure ,Apoptosis ,embryonic structures ,Cancer research ,Phosphorylation ,Original Article ,Casein kinase 2 ,Apoptosis Regulatory Proteins ,Par-4 - Abstract
International audience; The proapoptotic protein, prostate apoptosis response-4 (Par-4), acts as a tumor suppressor in prostate cancer cells. The serine/threonine kinase casein kinase 2 (CK2) has a well-reported role in prostate cancer resistance to apoptotic agents or anticancer drugs. However, the mechanistic understanding on how CK2 supports survival is far from complete. In this work, we demonstrate both in rat and humans that (i) Par-4 is a new substrate of the survival kinase CK2 and (ii) phosphorylation by CK2 impairs Par-4 proapoptotic functions. We also unravel different levels of CK2-dependent regulation of Par-4 between species. In rats, the phosphorylation by CK2 at the major site, S124, prevents caspase-mediated Par-4 cleavage (D123) and consequently impairs the proapoptotic function of Par-4. In humans, CK2 strongly impairs the apoptotic properties of Par-4, independently of the caspase-mediated cleavage of Par-4 (D131), by triggering the phosphorylation at residue S231. Furthermore, we show that human Par-4 residue S231 is highly phosphorylated in prostate cancer cells as compared with their normal counterparts. Finally, the sensitivity of prostate cancer cells to apoptosis by CK2 knockdown is significantly reversed by parallel knockdown of Par-4. Thus, Par-4 seems a critical target of CK2 that could be exploited for the development of new anticancer drugs.
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- 2014
17. Modulation of apoptosis by procaspase-2 short isoform: selective inhibition of chromatin condensation, apoptotic body formation and phosphatidylserine externalization
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Florence Bichat, Arlette Hammann, Eric Solary, Richard Bertrand, Cédric Rébé, and Nathalie Droin
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Cancer Research ,Programmed cell death ,Amino Acid Motifs ,Molecular Sequence Data ,Apoptosis ,DNA Fragmentation ,Phosphatidylserines ,Transfection ,chemistry.chemical_compound ,Tumor Cells, Cultured ,Genetics ,Humans ,Topoisomerase II Inhibitors ,Cysteine ,Enzyme Inhibitors ,Molecular Biology ,Caspase ,Etoposide ,Enzyme Precursors ,Leukemia ,Base Sequence ,biology ,Caspase 2 ,Phosphatidylserine ,Apoptotic body ,Molecular biology ,Chromatin ,Isoenzymes ,chemistry ,Caspases ,Mutation ,biology.protein ,DNA fragmentation ,Topoisomerase-II Inhibitor - Abstract
Procaspase-2 is one of the cysteine aspartate proteases involved in apoptotic cell death. Alternative splicing of CASP-2 messenger RNA generates a long isoform, procaspase-2L, whose overexpression induces cell death and a truncated isoform, procaspase-2S, whose function remains poorly defined. The present study explored the consequences of procaspase-2S overexpression in U937 human leukemic cells exposed to the topoisomerase II inhibitor etoposide as an apoptotic stimulus. Overexpression of procaspase-2S in U937 cells partially prevented nuclear changes associated with etoposide-induced cell death, as determined by Hoechst 33342 staining of nuclear chromatin and electron microscopy studies. Procaspase-2S also prevented the maturation of apoptotic bodies, delayed phosphatidylserine externalization on the plasma membrane and prevented the cleavage and activation of procaspase-2L. These effects were not observed when the cysteine 289 in the consensus QACRG motif was mutated into a serine. Wild-type procaspase-2S overexpression did not influence the cleavage of procaspase-3, procaspase-7 and poly(ADP-ribose)polymerase nor the fragmentation of nuclear DNA into nucleosome-sized fragments. Altogether, these results indicate that the short isoform of procaspase-2 negatively interferes with selective features of apoptosis, an activity that is suppressed by mutation of the cysteine 289.
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- 2001
18. HSP27 favors ubiquitination and proteasomal degradation of p27 Kip1 and helps S‐phase re‐entry in stressed cells
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Arnaud Parcellier, Mathilde Brunet, Elise Schmitt, Edwige Col, Celine Didelot, Arlette Hammann, Keiko Nakayama, Keiichi I. Nakayama, Saadi Khochbin, Eric Solary, Carmen Garrido, and Céline Didelot
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Proteasome Endopeptidase Complex ,Ubiquitin-Protein Ligases ,HSP27 Heat-Shock Proteins ,Resting Phase, Cell Cycle ,Biochemistry ,Culture Media, Serum-Free ,Cell Line ,S Phase ,Ubiquitin ,Hsp27 ,Cyclin-dependent kinase ,Genetics ,Animals ,Humans ,Phosphorylation ,S-Phase Kinase-Associated Proteins ,Molecular Biology ,Heat-Shock Proteins ,biology ,Cell growth ,Kinase ,G1 Phase ,Cell cycle ,Neoplasm Proteins ,Rats ,Cell biology ,Proteasome ,biology.protein ,Cyclin-Dependent Kinase Inhibitor p27 ,Molecular Chaperones ,Biotechnology - Abstract
Stress-inducible HSP27 protects cells from death through various mechanisms. We have recently demonstrated that HSP27 can also enhance the degradation of some proteins through the proteasomal pathway. Here, we show that one of these proteins is the cyclin-dependent kinase (Cdk) inhibitor p27Kip1. The ubiquitination and degradation of this protein that favors progression through the cell cycle was previously shown to involve either a Skp2-dependent mechanism,i.e., at the S-/G2-transition, or a KPC (Kip1 ubiquitination-promoting complex)-dependent mechanism, i.e.,at the G0/G1 transition. In this work, we demonstrate that, in response to serum depletion, p27Kip1 cellular content first increases then progressively decreases as cells begin to die. In this stressful condition, HSP27favors p27Kip1 ubiquitination and degradation by the proteasome. A similar observation was made in response to stress induced by the NO donor glyceryl trinitrate (GTN). HSP27-mediated ubiquitination ofp27Kip1 does not require its phosphorylation on Thr187 or Ser-10, nor does it depend on the SCFSkp2 ubiquitin ligase E3 complex. It facilitates the G1/S transition,which suggests that, in stressful conditions, HSP27might render quiescent cells competent to re-enter the cell cycle.
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- 2006
19. Inconstant Association between 27-kDa Heat-Shock Protein (Hsp27) Content and Doxorubicin Resistance in Human Colon Cancer Cells. The Doxorubicin-Protecting Effect of Hsp27
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Arlette Hammann, Bruno Chauffert, Annie Fromentin, Carmen Garrido, Mahfoud Assem, Patrick Mehlen, and André-Patrick Arrigo
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endocrine system ,Hot Temperature ,animal structures ,Anthracycline ,Cell Survival ,Drug Resistance ,Transfection ,Biochemistry ,Hsp27 ,Heat shock protein ,Tumor Cells, Cultured ,medicine ,Humans ,Doxorubicin ,RNA, Messenger ,Cytotoxicity ,Heat-Shock Proteins ,Cisplatin ,biology ,Cell cycle ,Molecular biology ,Gene Expression Regulation, Neoplastic ,Molecular Weight ,DNA Topoisomerases, Type II ,Colonic Neoplasms ,embryonic structures ,biology.protein ,Fluorouracil ,medicine.drug - Abstract
To investigate the role of the small 27-kDa heat-shock protein (Hsp27) in the intrinsic resistance of colon cancer cells to doxorubicin, we modified Hsp27 expression either genetically by transfection or pharmacologically by cisplatin treatment. HT-29 cells were transfected with a full-length Hsp27 construct in the sense or antisense orientation. We found a good correlation between cell survival after doxorubicin treatment and Hsp27 content. A similar correlation was found for the thermoresistance of the Hsp27-transfected cells. In contrast, the sensitivity of the different transfected cells to 5-fluorouracil was not modified. cis-Platinum(II)diammine dichloride (cisplatin) treatment of HT-29 or Caco2 cells dramatically increased their Hsp27 mRNA and protein content. Accordingly, the cells became thermoresistant. Contrary to what has been previously assumed, however, cell resistance to doxorubicin was reduced. Our data suggest that the decreased resistance of the cells to doxorubicin may be due to a concomitant increase of topoisomerase II expression, the main target of anthracyclines. In conclusion, although Hsp27 seems to participate in the natural resistance of colon cancer cells to anthracyclines, its increase after cisplatin treatment is not associated with a decreased cytotoxicity to doxorubicin.
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- 1996
20. Trim33/Tif1γ is involved in late stages of granulomonopoiesis in mice
- Author
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Laurent Delva, Arlette Hammann, Anne Largeot, Jean-Noël Bastie, Ronan Quéré, Malika Trad, Romain Aucagne, Romain Z. Martin, Caroline Legouge, Marie-Lorraine Chretien, Lipides - Nutrition - Cancer (U866) (LNC), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Bourgogne (UB)-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement, Laboratoire d'Excellence : Lipoprotéines et Santé : prévention et Traitement des maladies Inflammatoires et du Cancer (LabEx LipSTIC), Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Paris-Sud - Paris 11 (UP11)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Gustave Roussy (IGR)-Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy)-Université de Bourgogne (UB)-Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon)-Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] (UNICANCER/CRLCC-CGFL), UNICANCER-UNICANCER-Institut National de la Santé et de la Recherche Médicale (INSERM)-Fédération Francophone de la Cancérologie Digestive, FFCD-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université de Montpellier (UM), Service d'Hématologie Clinique (CHU de Dijon), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon), Processus d'Activation Sélective par Transfert d'Energie Uni-électronique ou Radiatif (UMR 8640) (PASTEUR), Université Pierre et Marie Curie - Paris 6 (UPMC)-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)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Plateforme cytométrie en flux [Fédération de Recherche Santé STIC de l’Université de Bourgogne], Université de Bourgogne (UB)-IFR100 - Structure fédérative de recherche Santé-STIC, Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC (UMR INSERM 1098) (RIGHT), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté]), Association pour la Recherche sur le Cancer (ARC), Ligue Nationale Contre le Cancer (Conference Inter-Regionale du Grand Est), association Cent pour Sang la Vie, Conseil Regional de Bourgogne (CRB) (PART), Societe Francaise d'Hematologie (SFH), ANR-11-LABX-0021,Lipstic,Lipoprotéines et santé : prévention et traitement des maladies inflammatoires non vasculaires et du(2011), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Gustave Roussy (IGR)-Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy)-Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Université de Bourgogne (UB)-Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon)-Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] (UNICANCER/CRLCC-CGFL), UNICANCER-UNICANCER-Institut National de la Santé et de la Recherche Médicale (INSERM)-Fédération Francophone de la Cancérologie Digestive, FFCD-Université de Montpellier (UM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS-PSL), Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Université de Franche-Comté (UFC), ANR-11-LABX-0021,Lipstic,Lipoprotéines et santé : prévention et traitement des maladies inflammatoires non vasculaires et du cancer(2011), Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Paris-Sud - Paris 11 (UP11)-École pratique des hautes études (EPHE)-Institut Gustave Roussy (IGR)-Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy)-Université de Bourgogne (UB)-Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon)-Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc (CRLCC - CGFL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Fédération Francophone de la Cancérologie Digestive, FFCD-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-France-Comté] (EFS [Bourgogne-France-Comté])-Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire [Besançon] (CHRU Besançon)-Université de Franche-Comté (UFC)-Université de Montpellier (UM), École normale supérieure - Paris (ENS Paris)-École normale supérieure - Paris (ENS Paris)-Centre National de la Recherche Scientifique (CNRS), Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC (UMR INSERM 1098) (HOTE GREFFON), Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang [Bourgogne-France-Comté] (EFS [Bourgogne-France-Comté])-Université de Franche-Comté (UFC), ANR-11-LABX-0021/11-LABX-0021,Lipstic,Lipoprotéines et santé : prévention et traitement des maladies inflammatoires non vasculaires et du(2011), Lipides - Nutrition - Cancer (U866) ( LNC ), Université de Bourgogne ( UB ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon ( ENSBANA ), Laboratoire d'Excellence : Lipoprotéines et Santé : prévention et Traitement des maladies Inflammatoires et du Cancer ( LabEx LipSTIC ), Institut National de la Recherche Agronomique ( INRA ) -Université Montpellier 2 - Sciences et Techniques ( UM2 ) -Université Paris-Sud - Paris 11 ( UP11 ) -École pratique des hautes études ( EPHE ) -Institut Gustave Roussy ( IGR ) -Centre Hospitalier Régional Universitaire de Nancy ( CHRU Nancy ) -Université de Bourgogne ( UB ) -Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand ( CHU Dijon ) -Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc ( CRLCC - CGFL ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Fédération Francophone de la Cancérologie Digestive, FFCD-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Etablissement français du sang [Bourgogne-France-Comté] ( EFS [Bourgogne-France-Comté] ) -Centre National de la Recherche Scientifique ( CNRS ) -Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Université de Franche-Comté ( UFC ), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand ( CHU Dijon ), Processus d'Activation Sélective par Transfert d'Energie Uni-électronique ou Radiatif (UMR 8640) ( PASTEUR ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Département de Chimie - ENS Paris, École normale supérieure - Paris ( ENS Paris ) -École normale supérieure - Paris ( ENS Paris ) -Centre National de la Recherche Scientifique ( CNRS ), Université de Bourgogne ( UB ) -IFR100 - Structure fédérative de recherche Santé-STIC, Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC ( HOTE GREFFON ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Etablissement français du sang [Bourgogne-France-Comté] ( EFS [Bourgogne-France-Comté] ) -Université de Franche-Comté ( UFC ), and ANR-11-LABX-0021/11-LABX-0021,Lipstic,Lipoprotéines et santé : prévention et traitement des maladies inflammatoires non vasculaires et du ( 2011 )
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0301 basic medicine ,Cancer Research ,Myeloid ,Apoptosis ,Trim33 ,Mice ,Cell Movement ,Granulocyte-Macrophage Progenitor Cells ,[ SDV.MHEP.HEM ] Life Sciences [q-bio]/Human health and pathology/Hematology ,Myeloid Cells ,Cancer ,Mice, Knockout ,Myelopoiesis ,Epithelial-cells ,Cell Differentiation ,[SDV.MHEP.HEM]Life Sciences [q-bio]/Human health and pathology/Hematology ,Hematology ,Ubiquitin ligase ,Haematopoiesis ,Phenotype ,medicine.anatomical_structure ,Integrin alpha M ,Stem cell ,Transcription ,Tumor-suppressor ,Chronic myelomonocytic leukemia ,Bone Marrow Cells ,Biology ,Immunophenotyping ,03 medical and health sciences ,Hepatocellular-carcinoma ,Genetics ,medicine ,Animals ,Cell Lineage ,Molecular Biology ,Myeloproliferative neoplasm ,Myeloproliferative Disorders ,Cell Biology ,Tif1-Gamma ,TRIM33 ,medicine.disease ,Tripartite motif family ,Suppressor-cells ,Disease Models, Animal ,030104 developmental biology ,Cancer research ,biology.protein ,Smad4 ,Biomarkers ,Transcription Factors - Abstract
IF 2.303; International audience; Trim33/Tif1γ (Trim33) is a member of the tripartite motif family. Using a conditional hematopoietic-specific Trim33 knock-out (Trim33(Δ/Δ)) mouse, we showed previously that Trim33 deficiency in hematopoietic stem cells leads to severe defects in hematopoiesis, resembling the main features of human chronic myelomonocytic leukemia. We also demonstrated that Trim33 is involved in hematopoietic aging through TGFβ signaling. Nevertheless, how Trim33 contributes to the terminal stages of myeloid differentiation remains to be clarified. We reveal here the crucial role of Trim33 expression in the control of mature granulomonocytic differentiation. An important component of Trim33-deficient mice is the alteration of myeloid differentiation, as characterized by dysplastic features, abnormal granulocyte and monocyte maturation, and the expansion of CD11b(+)Ly6G(high)Ly6C(low) myeloid cells, which share some features with polymorphonuclear-myeloid-derived suppressor cells. Moreover, in Trim33(Δ/Δ) mice, we observed the alteration of CSF-1-mediated macrophage differentiation in association with the lack of Csf-1 receptor. Altogether, these results indicate that Trim33 deficiency leads to the expansion of a subset of myeloid cells characterizing the myelodysplastic/myeloproliferative neoplasm.
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- 2016
21. Wee1 inhibition potentiates Wip1-dependent p53-negative tumor cell death during chemotherapy
- Author
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Anastasia R. Goloudina, Elena Y. Kochetkova, Marc Bardou, Arlette Hammann, Oleg N. Demidov, Sarah Richaud, Carmen Garrido, Olga A. Fedorova, Nickolai A. Barlev, Burhan Uyanik, V Clausse, Lipides - Nutrition - Cancer (U866) ( LNC ), Université de Bourgogne ( UB ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon ( ENSBANA ), Institute of Cytology of the Russian Academy of Science (St. Petersburg), Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc ( CRLCC - CGFL ), Fondation ARC, laboratoire d'excellence ARC, La Ligue Contre le Cancer CCIR-GE (14-15-00636 ), fondation scientifique russe, Lipides - Nutrition - Cancer (U866) (LNC), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA), Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] (UNICANCER/CRLCC-CGFL), and UNICANCER
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Wip1 ,Apoptosis ,Cell Cycle Proteins ,Pharmacology ,MESH: G2 Phase Cell Cycle Checkpoints ,Histones ,MESH : Phosphorylation ,Mice ,MESH : Cell Cycle Proteins ,MESH: Animals ,MESH: Tumor Suppressor Protein p53 ,MESH: Histones ,Kinase ,Tp53 mutations ,MESH : Mice, Transgenic ,3. Good health ,Protein Phosphatase 2C ,Survival Rate ,MESH : Antineoplastic Agents ,H2ax phosphorylation ,P53 activation ,MESH: Protein Phosphatase 2C ,RNA Interference ,MESH : Colorectal Neoplasms ,MESH : Carrier Proteins ,Histone H2ax ,MESH: Mitochondria ,Immunology ,Human fibroblasts ,MESH: Carrier Proteins ,Antineoplastic Agents ,MESH: Protein-Tyrosine Kinases ,MESH: Protein-Serine-Threonine Kinases ,MESH : Cisplatin ,03 medical and health sciences ,MESH: Cell Cycle Proteins ,Genotoxic stress ,MESH : Protein-Tyrosine Kinases ,Humans ,MESH : Histones ,Anticancer Therapy ,MESH: DNA Damage ,Cisplatin ,MESH: Humans ,MESH: Phosphorylation ,[ SDV.BC ] Life Sciences [q-bio]/Cellular Biology ,MESH : Humans ,MESH : Nuclear Proteins ,030104 developmental biology ,Cancer cell ,MESH: Antineoplastic Agents ,Carrier Proteins ,MESH: Nuclear Proteins ,MESH : Apoptosis ,Dna-damage response ,0301 basic medicine ,Cancer Research ,MESH: Caspase 3 ,MESH : Caspase 3 ,Phosphorylation ,Cytotoxicity ,MESH : DNA Damage ,Sensitization ,medicine.diagnostic_test ,Caspase 3 ,Nuclear Proteins ,Protein-Tyrosine Kinases ,MESH : Survival Rate ,Mitochondria ,G2 Phase Cell Cycle Checkpoints ,Wee1 ,medicine.anatomical_structure ,MESH : Protein Phosphatase 2C ,Original Article ,MESH : Mitochondria ,Colorectal Neoplasms ,medicine.drug ,MESH : Protein-Serine-Threonine Kinases ,MESH: Cell Line, Tumor ,MESH: Survival Rate ,MESH: Mice, Transgenic ,MESH: RNA Interference ,Phosphatase ,Mice, Transgenic ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,Biology ,Protein Serine-Threonine Kinases ,Flow cytometry ,Cellular and Molecular Neuroscience ,Cell Line, Tumor ,MESH : Mice ,medicine ,Animals ,MESH: Mice ,MESH : Cell Line, Tumor ,MESH: Apoptosis ,Cell Biology ,MESH : Tumor Suppressor Protein p53 ,MESH: Cisplatin ,Cancer research ,biology.protein ,MESH : Animals ,MESH : G2 Phase Cell Cycle Checkpoints ,MESH : RNA Interference ,Tumor Suppressor Protein p53 ,MESH: Colorectal Neoplasms ,DNA Damage - Abstract
Inactivation of p53 found in more than half of human cancers is often associated with increased tumor resistance to anti-cancer therapy. We have previously shown that overexpression of the phosphatase Wip1 in p53-negative tumors sensitizes them to chemotherapeutic agents, while protecting normal tissues from the side effects of anti-cancer treatment. In this study, we decided to search for kinases that prevent Wip1-mediated sensitization of cancer cells, thereby interfering with efficacy of genotoxic anti-cancer drugs. To this end, we performed a flow cytometry-based screening in order to identify kinases that regulated the levels of γH2AX, which were used as readout. Another criterion of the screen was increased sensitivity of p53-negative tumor cells to cisplatin (CDDP) in a Wip1-dependent manner. We have found that a treatment with a low dose (75 nM) of MK-1775, a recently described specific chemical inhibitor of Wee1, decreases CDDP-induced H2AX phosphorylation in p53-negative cells and enhances the Wip1-sensitization of p53-negative tumors. We were able to reduce CDDP effective concentration by 40% with a combination of Wip1 overexpression and Wee1 kinase inhibition. We have observed that Wee1 inhibition potentiates Wip1-dependent tumor sensitization effect by reducing levels of Hipk2 kinase, a negative regulator of Wip1 pathway. In addition, during CDDP treatment, the combination of Wee1 inhibition and Wip1 overexpression has a mild but significant protective effect in normal cells and tissues. Our results indicate that inhibition of the negative regulators of Wip1 pathway, Wee1 and Hipk2, in p53-negative tumors could potentiate efficiency of chemotherapeutic agents without concomitant increase of cytotoxicity in normal tissues. The development and clinical use of Wee1 and Hipk1 kinase chemical inhibitors might be a promising strategy to improve anti-cancer therapy.
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- 2016
22. Wip1 promotes RUNX2-dependent apoptosis in p53-negative tumors and protects normal tissues during treatment with anticancer agents
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Sharlyn J. Mazur, Dmitry V. Bulavin, Ettore Appella, Xavier Le Guezennec, Oleg N. Demidov, Carmen Garrido, Arlette Hammann, Eric Fourmaux, Anastasia R. Goloudina, and Kan Tanoue
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Blotting, Western ,Phosphatase ,Antineoplastic Agents ,Apoptosis ,Core Binding Factor Alpha 1 Subunit ,Biology ,Pyruvate dehydrogenase phosphatase ,Real-Time Polymerase Chain Reaction ,Cell Line ,Mice ,Bcl-2-associated X protein ,Neoplasms ,Phosphoprotein Phosphatases ,Animals ,Humans ,Immunoprecipitation ,DNA Primers ,bcl-2-Associated X Protein ,Regulation of gene expression ,Multidisciplinary ,Reverse Transcriptase Polymerase Chain Reaction ,Drug Synergism ,Immunohistochemistry ,Gene Expression Regulation, Neoplastic ,Protein Phosphatase 2C ,Blot ,PNAS Plus ,Cell culture ,Cancer research ,biology.protein ,Female ,Tumor Suppressor Protein p53 ,Plasmids - Abstract
The inactivation of the p53 tumor suppressor pathway in many cancers often increases their resistance to anticancer therapy. Here we show that a previously proposed strategy directed to Wip1 inhibition could be ineffective in tumors lacking p53. On the contrary, Wip1 overexpression sensitized these tumors to chemotherapeutic agents. This effect was mediated through interaction between Wip1 and RUNX2 that resulted, in response to anticancer treatment, in RUNX2-dependent transcriptional induction of the proapoptotic Bax protein. The potentiating effects of Wip1 overexpression on chemotherapeutic agents were directed only to tumor cells lacking p53. The overexpression of Wip1 in normal tissues provided protection from cisplatin-induced apoptosis through decreased strength of upstream signaling to p53. Thus, Wip1 phosphatase promotes apoptosis in p53-negative tumors and protects normal tissues during treatment with anticancer agents.
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- 2011
23. Cellular Inhibitor of Apoptosis Protein-1 (cIAP1) Can Regulate E2F1 Transcription Factor-mediated Control of Cyclin Transcription
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Arlette Hammann, Beatrice Eymin, Jean Berthelet, Arthur Marivin, Eric Solary, Laurence Dubrez, Laurent Delva, Valérie Edmond, Brice Lagrange, Alban Dupoux, Stéphanie Plenchette, Jessy Cartier, Simon Gemble, Lipides - Nutrition - Cancer (U866) (LNC), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA), Equipe 2 - Bases Moléculaires de la Progression des Cancers du Poumon, Inserm U823, Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), and Eymin, Beatrice
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Transcription, Genetic ,Cellular differentiation ,[SDV]Life Sciences [q-bio] ,Cyclin A ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,Response Elements ,Inhibitor of apoptosis ,Biochemistry ,Inhibitor of Apoptosis Proteins ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Cyclin E ,Animals ,Humans ,E2F1 ,Gene Silencing ,E2F ,[SDV.BC] Life Sciences [q-bio]/Cellular Biology ,Molecular Biology ,Cell Proliferation ,030304 developmental biology ,Cell Nucleus ,0303 health sciences ,biology ,E2F1 Transcription Factor ,Cell Biology ,Cell cycle ,Molecular biology ,Protein Structure, Tertiary ,3. Good health ,Cell biology ,[SDV] Life Sciences [q-bio] ,030220 oncology & carcinogenesis ,biology.protein ,biological phenomena, cell phenomena, and immunity ,Chromatin immunoprecipitation ,HeLa Cells - Abstract
International audience; The inhibitor of apoptosis protein cIAP1 (cellular inhibitor of apoptosis protein-1) is a potent regulator of the tumor necrosis factor (TNF) receptor family and NF-B signaling pathways in the cytoplasm. However, in some primary cells and tumor cell lines, cIAP1 is expressed in the nucleus, and its nuclear function remains poorly understood. Here, we show that the N-terminal part of cIAP1 directly interacts with the DNA binding domain of the E2F1 transcription factor. cIAP1 dramatically increases the transcriptional activity of E2F1 on synthetic and CCNE promoters. This function is not conserved for cIAP2 and XIAP, which are cytoplasmic proteins. Chromatin immunoprecipitation experiments demonstrate that cIAP1 is recruited on E2F binding sites of the CCNE and CCNA promoters in a cell cycle-and differentiation-dependent manner. cIAP1 silencing inhibits E2F1 DNA binding and E2F1-mediated transcriptional activation of the CCNE gene. In cells that express a nuclear cIAP1 such as HeLa, THP1 cells and primary human mammary epithelial cells, down-regulation of cIAP1 inhibits cyclin E and A expression and cell proliferation. We conclude that one of the functions of cIAP1 when localized in the nucleus is to regulate E2F1 transcriptional activity.
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- 2011
24. Peptides and Aptamers Targeting HSP70: A Novel Approach for Anticancer Chemotherapy
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Anne-Laure, Rérole, Jessica, Gobbo, Aurelie, De Thonel, Elise, Schmitt, Jean Paul, Pais de Barros, Arlette, Hammann, David, Lanneau, Eric, Fourmaux, Oleg N, Demidov, Oleg, Deminov, Olivier, Micheau, Laurent, Lagrost, Pierre, Colas, Guido, Kroemer, Carmen, Garrido, Centre épigénétique et destin cellulaire (EDC (UMR_7216)), and Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP)
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Cancer Research ,Aptamer ,[SDV]Life Sciences [q-bio] ,Melanoma, Experimental ,Peptide ,Antineoplastic Agents ,Apoptosis ,Transfection ,03 medical and health sciences ,Mice ,0302 clinical medicine ,In vivo ,Heat shock protein ,medicine ,Animals ,Humans ,HSP70 Heat-Shock Proteins ,Molecular Targeted Therapy ,ComputingMilieux_MISCELLANEOUS ,030304 developmental biology ,chemistry.chemical_classification ,0303 health sciences ,Chemistry ,Melanoma ,medicine.disease ,3. Good health ,Hsp70 ,Protein Structure, Tertiary ,Rats ,Mice, Inbred C57BL ,Oncology ,Biochemistry ,030220 oncology & carcinogenesis ,Cancer research ,Peptides ,Aptamers, Peptide ,HeLa Cells - Abstract
The inhibition of heat shock protein 70 (HSP70) is an emerging strategy in cancer therapy. Unfortunately, no specific inhibitors are clinically available. By yeast two-hybrid screening, we have identified multiple peptide aptamers that bind HSP70. When expressed in human tumor cells, two among these peptide aptamers—A8 and A17—which bind to the peptide-binding and the ATP-binding domains of HSP70, respectively, specifically inhibited the chaperone activity, thereby increasing the cells' sensitivity to apoptosis induced by anticancer drugs. The 13-amino acid peptide from the variable region of A17 (called P17) retained the ability to specifically inhibit HSP70 and induced the regression of subcutaneous tumors in vivo after local or systemic injection. This antitumor effect was associated with an important recruitment of macrophages and T lymphocytes into the tumor bed. Altogether, these data indicate that peptide aptamers or peptides that target HSP70 may be considered as novel lead compounds for cancer therapy. Cancer Res; 71(2); 484–95. ©2011 AACR.
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- 2011
25. Involvement of T lymphocytes in curative effect of a new immunomodulator OM 163 on rat colon cancer metastases
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Nathalie Onier, Pierre Hirt, Jean-François Jeannin, Arlette Hammann, Patricia Lagadec, Monique Martin, Pascale Lejeune, and Jacques Bauer
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Male ,Cancer Research ,Pathology ,medicine.medical_specialty ,Lung Neoplasms ,Time Factors ,Colorectal cancer ,T-Lymphocytes ,medicine.medical_treatment ,Dose-Response Relationship, Immunologic ,Metastasis ,Rats, Nude ,Adjuvants, Immunologic ,Escherichia coli ,medicine ,Splenocyte ,Animals ,Peritoneal Neoplasms ,Curative effect ,Lung ,business.industry ,Immunotherapy ,medicine.disease ,Rats, Inbred F344 ,Rats ,Peritoneal carcinomatosis ,medicine.anatomical_structure ,Oncology ,Colonic Neoplasms ,Female ,business ,CD8 - Abstract
In a model of colon cancer in syngeneic rats, a new immunomodulator, OM 163, induced the complete disappearance of peritoneal carcinomatosis (nodules measuring 1–5 mm) in 41 out of 82 rats. Those results were confirmed in a survival experiment in which 3 out of 10 treated rats died free of tumour 10, 18 and 28 months after the tumour cell injection while all the untreated control rats died of their tumours within 3 months. OM 163 had a systemic effect, since injected intraperitoneally it completely inhibited the growth of lung metastases in 13 out of 20 rats. The antitumour effect of OM 163 was also observed in two rat strains on original tumours. Lymphocyte infiltration was observed in the tumours mainly constituted of CD4 + and CD8 + cells. The treatment had no effect in nude rats, confirming the involvement of T lymphocytes. Furthermore, rats cured by OM 163 were protected against a second challenge of tumour cells and in a Winn's assay, splenocytes from cured rats protected normal rats against tumour cells.
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- 1993
26. Transactivation of the epidermal growth factor receptor by heat shock protein 90 via Toll-like receptor 4 contributes to the migration of glioblastoma cells
- Author
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Dominique Thuringer, Naïma Benikhlef, Arlette Hammann, Guillaume Wettstein, Eric Solary, Carmen Garrido, Eric Fourmaux, and André Bouchot
- Subjects
Transcriptional Activation ,Biology ,Biochemistry ,Adenosine Triphosphate ,Membrane Microdomains ,Cell surface receptor ,Cell Movement ,Heat shock protein ,Cell Line, Tumor ,Extracellular ,Humans ,Calcium Signaling ,HSP90 Heat-Shock Proteins ,Protein kinase A ,Molecular Biology ,Apyrase ,Purinergic receptor ,Cell migration ,Cell Biology ,Cell biology ,ErbB Receptors ,Toll-Like Receptor 4 ,Protein Kinase C-delta ,Signal transduction ,Glioblastoma ,Signal Transduction - Abstract
Extracellular heat shock protein HSP90α was reported to participate in tumor cell growth, invasion, and metastasis formation through poorly understood signaling pathways. Herein, we show that extracellular HSP90α favors cell migration of glioblastoma U87 cells. More specifically, externally applied HSP90α rapidly induced endocytosis of EGFR. This response was accompanied by a transient increase in cytosolic Ca(2+) appearing after 1-3 min of treatment. In the presence of EGF, U87 cells showed HSP90α-induced Ca(2+) oscillations, which were reduced by the ATP/ADPase, apyrase, and inhibited by the purinergic P(2) inhibitor, suramin, suggesting that ATP release is requested. Disruption of lipid rafts with methyl β-cyclodextrin impaired the Ca(2+) rise induced by extracellular HSP90α combined with EGF. Specific inhibition of TLR4 expression by blocking antibodies suppressed extracellular HSP90α-induced Ca(2+) signaling and the associated cell migration. HSPs are known to bind lipopolysaccharides (LPSs). Preincubating cells with Polymyxin B, a potent LPS inhibitor, partially abrogated the effects of HSP90α without affecting Ca(2+) oscillations observed with EGF. Extracellular HSP90α induced EGFR phosphorylation at Tyr-1068, and this event was prevented by both the protein kinase Cδ inhibitor, rottlerin, and the c-Src inhibitor, PP2. Altogether, our results suggest that extracellular HSP90α transactivates EGFR/ErbB1 through TLR4 and a PKCδ/c-Src pathway, which induces ATP release and cytosolic Ca(2+) increase and finally favors cell migration. This mechanism could account for the deleterious effects of HSPs on high grade glioma when released into the tumor cell microenvironment.
- Published
- 2010
27. Transcription intermediary factor 1γ is a tumor suppressor in mouse and human chronic myelomonocytic leukemia
- Author
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Nathalie Droin, Anne Largeot, Eric Solary, Jérôme Paggetti, Jean-Noël Bastie, Arlette Hammann, Kai-Ping Yan, Romain Aucagne, Amandine Bataille, Laurent Delva, Pierre Fenaux, Brice Lagrange, Régine Losson, and Laurent Martin
- Subjects
Male ,Aging ,Antimetabolites, Antineoplastic ,Tumor suppressor gene ,Cellular differentiation ,Molecular Sequence Data ,Chronic myelomonocytic leukemia ,Receptor, Macrophage Colony-Stimulating Factor ,Biology ,Decitabine ,chemistry.chemical_compound ,Mice ,medicine ,Animals ,Humans ,Genes, Tumor Suppressor ,Promoter Regions, Genetic ,Transcription factor ,Aged ,Aged, 80 and over ,Mice, Knockout ,Base Sequence ,Gene Expression Regulation, Leukemic ,Cell Differentiation ,Leukemia, Myelomonocytic, Chronic ,General Medicine ,DNA Methylation ,Middle Aged ,medicine.disease ,TRIM33 ,Hematopoietic Stem Cells ,Molecular biology ,Demethylating agent ,Hematopoiesis ,Neoplasm Proteins ,Specific Pathogen-Free Organisms ,Haematopoiesis ,chemistry ,DNA methylation ,Cancer research ,Azacitidine ,Female ,Transcription Factors ,Research Article - Abstract
Transcription intermediary factor 1γ (TIF1γ) was suggested to play a role in erythropoiesis. However, how TIF1γ regulates the development of different blood cell lineages and whether TIF1γ is involved in human hematological malignancies remain to be determined. Here we have shown that TIF1γ was a tumor suppressor in mouse and human chronic myelomonocytic leukemia (CMML). Loss of Tif1g in mouse HSCs favored the expansion of the granulo-monocytic progenitor compartment. Furthermore, Tif1g deletion induced the age-dependent appearance of a cell-autonomous myeloproliferative disorder in mice that recapitulated essential characteristics of human CMML. TIF1γ was almost undetectable in leukemic cells of 35% of CMML patients. This downregulation was related to the hypermethylation of CpG sequences and specific histone modifications in the gene promoter. A demethylating agent restored the normal epigenetic status of the TIF1G promoter in human cells, which correlated with a reestablishment of TIF1γ expression. Together, these results demonstrate that TIF1G is an epigenetically regulated tumor suppressor gene in hematopoietic cells and suggest that changes in TIF1γ expression may be a biomarker of response to demethylating agents in CMML.
- Published
- 2010
28. HSP27 controls GATA-1 protein level during erythroid cell differentiation
- Author
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Subramaniam Selvakumar, Joan Boyes, Olivier Hermine, Arlette Hammann, David Lanneau, Mathilde Brunet, Geneviève Courtois, Eric Solary, Julie Vandekerckhove, Aurélie de Thonel, Anne Sophie Gabet, J.-A. Ribeil, Yael Zermati, Sebastien Maurel, Adonis Hazoumé, Carmen Garrido, Cytokines, hématopoïèse et réponse immune (CHRI), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut International de Paléoprimatologie, Paléontologie Humaine : Evolution et Paléoenvironnement (IPHEP), Université de Poitiers-Centre National de la Recherche Scientifique (CNRS), Groupe français des myéloplastes and groupe ouest est des leucemies aigües myéloïde (SERVICE DES MALADIES DU SANG), Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), Instituto Andaluz de Ciencias de la Tierra (IACT), Consejo Superior de Investigaciones Científicas [Spain] (CSIC)-Universidad de Granada (UGR), Slama, Catherine, Centre épigénétique et destin cellulaire (EDC (UMR_7216)), Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Consejo Superior de Investigaciones Científicas [Madrid] (CSIC)-Universidad de Granada (UGR), Cytokines, hématopoïèse et réponse immune ( CHRI ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Institut International de Paléoprimatologie, Paléontologie Humaine : Evolution et Paléoenvironnement ( IPHEP ), Université de Poitiers-Centre National de la Recherche Scientifique ( CNRS ), Groupe français des myéloplastes and groupe ouest est des leucemies aigües myéloïde ( SERVICE DES MALADIES DU SANG ), Centre Hospitalier Régional Universitaire [Lille] ( CHRU Lille ), Instituto Andaluz de Ciencias de la Tierra ( IACT ), and Consejo Superior de Investigaciones Científicas [Spain] ( CSIC ) -Universidad de Granada ( UGR )
- Subjects
Leupeptins ,Pyridines ,[SDV]Life Sciences [q-bio] ,Cellular differentiation ,Cell ,HSP27 Heat-Shock Proteins ,Antigens, CD34 ,Biochemistry ,p38 Mitogen-Activated Protein Kinases ,0302 clinical medicine ,Transforming Growth Factor beta ,hemic and lymphatic diseases ,Chlorocebus aethiops ,GATA1 Transcription Factor ,Phosphorylation ,ComputingMilieux_MISCELLANEOUS ,Cells, Cultured ,Heat-Shock Proteins ,0303 health sciences ,biology ,Imidazoles ,Cell Differentiation ,Hematology ,[SDV] Life Sciences [q-bio] ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,embryonic structures ,COS Cells ,RNA Interference ,Signal transduction ,Proteasome Inhibitors ,Protein Binding ,Proteasome Endopeptidase Complex ,Immunology ,Immunoblotting ,03 medical and health sciences ,Hsp27 ,Erythroid Cells ,Heat shock protein ,medicine ,Animals ,Humans ,Transcription factor ,030304 developmental biology ,Cell Nucleus ,Interleukin-6 ,Ubiquitination ,Cell Biology ,Transforming growth factor beta ,Molecular biology ,Chaperone (protein) ,biology.protein ,K562 Cells ,HeLa Cells ,Molecular Chaperones - Abstract
Heat shock protein 27 (HSP27) is a chaperone whose cellular expression increases in response to various stresses and protects the cell either by inhibiting apoptotic cell death or by promoting the ubiquitination and proteasomal degradation of specific proteins. Here, we show that globin transcription factor 1 (GATA-1) is a client protein of HSP27. In 2 models of erythroid differentiation; that is, in the human erythroleukemia cell line, K562 induced to differentiate into erythroid cells on hemin exposure and CD34+ human cells ex vivo driven to erythroid differentiation in liquid culture, depletion of HSP27 provokes an accumulation of GATA-1 and impairs terminal maturation. More specifically, we demonstrate that, in the late stages of the erythroid differentiation program, HSP27 is phosphorylated in a p38-dependent manner, enters the nucleus, binds to GATA-1, and induces its ubiquitination and proteasomal degradation, provided that the transcription factor is acetylated. We conclude that HSP27 plays a role in the fine-tuning of terminal erythroid differentiation through regulation of GATA-1 content and activity.
- Published
- 2010
29. Histological reactivity of a monoclonal antibody against rat colon cancer cells on human and rat normal gut and colonic tumours
- Author
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Rafael Oriol, Arlette Hammann, Philippe Genne, François Martin, Nils-Olivier Olsson, Anne Caignard, and J. Bara
- Subjects
Male ,Pathology ,medicine.medical_specialty ,Colorectal cancer ,medicine.drug_class ,Molecular Sequence Data ,Immunocytochemistry ,Oligosaccharides ,Adenocarcinoma ,Biology ,Monoclonal antibody ,Epitope ,Pathology and Forensic Medicine ,Immunoenzyme Techniques ,Epitopes ,symbols.namesake ,Carbohydrate Conformation ,medicine ,Animals ,Humans ,Intestinal Mucosa ,Molecular Biology ,Dimethylhydrazines ,chemistry.chemical_classification ,Human gastrointestinal tract ,Antibodies, Monoclonal ,Rats, Inbred Strains ,Cell Biology ,General Medicine ,Immunogold labelling ,Golgi apparatus ,medicine.disease ,1,2-Dimethylhydrazine ,Rats ,medicine.anatomical_structure ,Carbohydrate Sequence ,chemistry ,Gastric Mucosa ,Organ Specificity ,Colonic Neoplasms ,Blood Group Antigens ,Carcinogens ,symbols ,Female ,Glycoprotein ,Fluorescein-5-isothiocyanate - Abstract
A monoclonal antibody, F11C, was raised against rat colon cancer cells. Its immunoreactivity on normal human and rat gut as well as human and rat colonic tumours was studied by the avidin-biotin-peroxidase complex technique. In both normal rat and human gastrointestinal tract, F11C stained surface epithelial cells from the fundus to distal colon, mainly as supranuclear vesicles. These vesicles appeared to be part of the Golgi apparatus on electron microscopy with immunogold labelling. Twenty primary rat colon tumours and 28 of 43 human colon tumours were also stained, with a heterogeneous pattern but much more strongly than the normal colonic mucosa. Biochemical purification suggested that in rat tumours F11C epitope was carried by a high molecular weight glycoprotein. Absorption experiments with synthetic oligosaccharides showed that F11C monoclonal antibody reacted with blood group A-related oligosaccharides. Nevertheless, F11C reactivity on human tissues was not related to the individual ABO or Lewis phenotype.
- Published
- 1992
30. Heat shock protein 27 is involved in SUMO-2/3 modification of heat shock factor 1 and thereby modulates the transcription factor activity
- Author
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Guillaume Bossis, M Brunet Simioni, Jacques Landry, Carmen Garrido, A de Thonel, Marc Piechaczyk, A Bouchot, Eric Fourmaux, Arlette Hammann, Anne-Laure Joly, Institut de Génétique Moléculaire de Montpellier (IGMM), and Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)
- Subjects
Protein sumoylation ,Transcriptional Activation ,Cancer Research ,endocrine system ,animal structures ,SUMO protein ,HSP27 Heat-Shock Proteins ,Biology ,urologic and male genital diseases ,environment and public health ,Substrate Specificity ,03 medical and health sciences ,Transactivation ,0302 clinical medicine ,Heat Shock Transcription Factors ,Heat shock protein ,Genetics ,Animals ,Humans ,Animals Cell Nucleus/metabolism DNA-Binding Proteins/*metabolism HSP27 Heat-Shock Proteins/chemistry/*metabolism Hela Cells Humans Protein Multimerization Protein Structure ,[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology ,HSF1 ,Protein Structure, Quaternary ,Molecular Biology ,Transcription factor ,Ubiquitins ,Heat-Shock Proteins ,030304 developmental biology ,Cell Nucleus ,0303 health sciences ,Molecular biology ,Hsp70 ,Cell biology ,Heat shock factor ,DNA-Binding Proteins ,Protein Transport ,Quaternary Protein Transport Small Ubiquitin-Related Modifier Proteins/*metabolism Substrate Specificity Transcription Factors/*metabolism Transcriptional Activation Ubiquitins/*metabolism ,030220 oncology & carcinogenesis ,embryonic structures ,Small Ubiquitin-Related Modifier Proteins ,Protein Multimerization ,HeLa Cells ,Molecular Chaperones ,Transcription Factors - Abstract
Heat shock protein 27 (HSP27) accumulates in stressed cells and helps them to survive adverse conditions. We have already shown that HSP27 has a function in the ubiquitination process that is modulated by its oligomerization/phosphorylation status. Here, we show that HSP27 is also involved in protein sumoylation, a ubiquitination-related process. HSP27 increases the number of cell proteins modified by small ubiquitin-like modifier (SUMO)-2/3 but this effect shows some selectivity as it neither affects all proteins nor concerns SUMO-1. Moreover, no such alteration in SUMO-2/3 conjugation is achievable by another HSP, such as HSP70. Heat shock factor 1 (HSF1), a transcription factor responsible for HSP expression, is one of the targets of HSP27. In stressed cells, HSP27 enters the nucleus and, in the form of large oligomers, binds to HSF1 and induces its modification by SUMO-2/3 on lysine 298. HSP27-induced HSF1 modification by SUMO-2/3 takes place downstream of the transcription factor phosphorylation on S303 and S307 and does not affect its DNA-binding ability. In contrast, this modification blocks HSF1 transactivation capacity. These data show that HSP27 exerts a feedback inhibition of HSF1 transactivation and enlighten the strictly regulated interplay between HSPs and HSF1. As we also show that HSP27 binds to the SUMO-E2-conjugating enzyme, Ubc9, our study raises the possibility that HSP27 may act as a SUMO-E3 ligase specific for SUMO-2/3.
- Published
- 2009
31. Growth characteristics and metastatic potential of seven intestinal carcinoma lines serially passaged in syngeneic rats
- Author
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Arlette Hammann, M. F. Michel, Monique Martin, and François Martin
- Subjects
Pathology ,medicine.medical_specialty ,Epithelioma ,Carcinoma ,Mucin ,Rats, Inbred Strains ,Cell Biology ,General Medicine ,Biology ,medicine.disease ,Rats ,Pathology and Forensic Medicine ,Metastasis ,Transplantation ,Transplantation, Isogeneic ,Intestinal Neoplasms ,Tumor Cells, Cultured ,Dimethylhydrazine ,medicine ,Homologous chromosome ,Animals ,Duodenal Carcinoma ,Molecular Biology ,Neoplasm Transplantation - Abstract
Transplantable tumour lines were obtained from one duodenal carcinoma induced by N-methyl-N'-nitro-N-nitrosoguanidine in the Lewis rat and from six colonic carcinomas induced by 1,2 dimethylhydrazine in BDIX or Fisher rats. The tumours were serially transplanted by the subcutaneous route into homologous syngeneic rats. The seven tumours differ from one another in their histological structure, five of them being well or moderately differentiated adenocarcinomas, and in their capacity to produce neutral or acidic mucins. The seven tumours also differ in their growth rate. The seven lines produced metastases; the metastatic potential and the location of the metastases differed from one line to another. The seven lines kept their original differentiation characteristics through multiple passages, representing several years of transplantation into syngeneic hosts. These tumours represent a useful and diversified model of metastatic intestinal carcinoma, available for basic research and therapeutic trials.
- Published
- 1991
32. MOZ/TIF2-induced acute myeloid leukaemia in transgenic fish
- Author
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Julia Zhuravleva, Arlette Hammann, Laurent Delva, Jérôme Paggetti, Jean-Noël Bastie, Eric Solary, and Laurent Martin
- Subjects
Myeloid ,Microinjections ,Oncogene Proteins, Fusion ,Transgene ,Biology ,Kidney ,MYST3 ,Fusion gene ,Animals, Genetically Modified ,Nuclear Receptor Coactivator 2 ,hemic and lymphatic diseases ,medicine ,Animals ,Zebrafish ,Gene ,Histone Acetyltransferases ,SPI1 ,Reverse Transcriptase Polymerase Chain Reaction ,Hematology ,medicine.disease ,biology.organism_classification ,Molecular biology ,Leukemia ,Disease Models, Animal ,Leukemia, Myeloid, Acute ,medicine.anatomical_structure ,embryonic structures ,Cancer research ,Gene Fusion - Abstract
The inv(8)(p11q13) chromosomal abnormality, described in acute myeloid leukaemias (AML), fuses the histone acetyl-transferase (HAT) MYST3 (MOZ) gene with another HAT gene, NCOA2 (TIF2). We generated a transgenic zebrafish in which the MYST3/NCOA2 fusion gene was expressed under control of the spi1 promoter. An AML developed in 2 of 180 MYST3/NCOA2-EGFP-expressing embryos, 14 and 26 months after injection of the fusion gene in a one-cell embryo, respectively. This leukaemia was characterised by an extensive invasion of kidneys by myeloid blast cells. This model, which is the first zebrafish model of AML, demonstrates the oncogenic potency of MYST3/NCOA2 fusion gene.
- Published
- 2008
33. Analysis of CD36 expression on human monocytic cells and atherosclerotic tissue sections with quantum dots: investigation by flow cytometry and spectral imaging microscopy
- Author
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Edmond, Kahn, Anne, Vejux, Franck, Ménétrier, Christophe, Maiza, Arlette, Hammann, Anabelle, Sequeira-Le Grand, Frédérique, Frouin, Yves, Tourneur, Frédéric, Brau, Jean-Marc, Riedinger, Eric, Steinmetz, Andrew, Todd-Pokropek, and Gérard, Lizard
- Subjects
CD36 Antigens ,Microscopy, Confocal ,Quantum Dots ,Image Processing, Computer-Assisted ,Fluorescent Antibody Technique ,Humans ,Phycoerythrin ,Arteries ,U937 Cells ,Atherosclerosis ,Flow Cytometry ,Fluorescein-5-isothiocyanate ,Monocytes - Abstract
To demonstrate CD36 expression with quantum dots (QDs) 525 and/or 605 on human monocytic U937 cells and atherosclerotic tissue sections by means of flow cytometry (FCM) and/or confocal laser scanning microscopy (CLSM).U937 cells and tissue sections were analyzed by means of FCM and/or CLSM. FCM was performed, using different ultraviolet (UV) and visible (488/532 nm) excitation modes. In the visible mode, fluorescence intensities of QDs, phycoerythrin (PE) and fluorescein isothiocyanate (FITC) were compared. Three-dimensional (3-D) sequences of images were obtained by spectral analysis in a CLSM and analyzed by the factor analysis of medical image sequences (FAMIS) algorithm, providing factor curves and images. Factor images are the result of the FAMIS image processing method, which differentiates emission spectra from 3D sequences of images. In CLSM analysis, preparations are screened in a UV excitation mode to optimize the possibilities of QDs and have the benefit of 4',6-diamino-2-phenylindole or Hoechst 33342 counterstaining of nuclei.FCM and CLSM revealed CD36 expression by means of QDs 525 and/or 605. Fluorescence intensity of PE and of FITC was higher than that of QDs 525 and of 605. As factor curves and images show the red emission of QDs 605 only, subsequent reliable identification and localization of CD36 was obtained.QDs 525 and 605 are useful to analyze antigenic expression. Following FCM, which is well adapted to detect fluorescence emission of QDs in the UV or visible excitation mode, CLSM and subsequent spectral analysis assess more specific characterization of QD fluorescent emissions.
- Published
- 2006
34. Small heat shock proteins HSP27 and alphaB-crystallin: cytoprotective and oncogenic functions
- Author
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Arnaud Parcellier, Mathilde Brunet, Arlette Hammann, Carmen Garrido, Elise Schmitt, and Eric Solary
- Subjects
Programmed cell death ,Proteasome Endopeptidase Complex ,Physiology ,Cell Survival ,Upstream and downstream (transduction) ,Clinical Biochemistry ,Apoptosis ,Biochemistry ,Hsp27 ,Heat shock protein ,Neoplasms ,medicine ,Humans ,Molecular Biology ,Heat-Shock Proteins ,General Environmental Science ,biology ,alpha-Crystallin B Chain ,Cell Biology ,Hsp70 ,Cell biology ,Heat shock factor ,Shock (circulatory) ,biology.protein ,General Earth and Planetary Sciences ,Protein folding ,medicine.symptom - Abstract
Heat shock protein-27 (HSP27) and alphaB-crystallin are ubiquitous small heat shock proteins whose expression is induced in response to a wide variety of physiological and environmental insults. They allow the cells to survive in otherwise lethal conditions. Various mechanisms have been proposed to account for the cytoprotective functions of these small heat shock proteins. First, these proteins are powerful molecular chaperones whose main function is to prevent the aggregation of nascent and stress-accumulated misfolded proteins. Second, they interact directly with various components of the tightly regulated programmed cell death machinery, upstream and downstream of the mitochondrial events. Third, they appear to play a role in the proteasome-mediated degradation of selected proteins. Both HSP27 and alphaB-crystallin were also proposed to participate in the development of neurodegenerative diseases and malignant tumors in which their overexpression could induce drug resistance. Altogether, these properties suggest that these small heat shock proteins are appropriate targets for modulating cell death pathways.
- Published
- 2005
35. Cisplatin-induced CD95 redistribution into membrane lipid rafts of HT29 human colon cancer cells
- Author
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Marie-Thérèse Dimanche-Boitrel, Sandrine Lacour, Eric Solary, Arlette Hammann, Odile Sergent, Solène Grazide, Philippe Gambert, Guy Laurent, Anne Athias, and Dominique Lagadic-Gossmann
- Subjects
Cancer Research ,Programmed cell death ,Ceramide ,Nystatin ,Membrane Fluidity ,Antineoplastic Agents ,Apoptosis ,Biology ,Ceramides ,chemistry.chemical_compound ,Membrane Microdomains ,medicine ,Membrane fluidity ,Humans ,Drug Interactions ,fas Receptor ,Lipid raft ,Cisplatin ,Plasma membrane raft ,Cell biology ,Sphingomyelin Phosphodiesterase ,Oncology ,Biochemistry ,chemistry ,biological phenomena, cell phenomena, and immunity ,Acid sphingomyelinase ,HT29 Cells ,medicine.drug - Abstract
We have shown previously that the death receptor CD95 could contribute to anticancer drug-induced apoptosis of colon cancer cells. In addition, anticancer drugs cooperate with CD95 cognate ligand or agonistic antibodies to trigger cancer cell apoptosis. In the present study, we show that the anticancer drug cisplatin induces clustering of CD95 at the surface of the human colon cancer cell line HT29, an event inhibited by the inhibitor of acid sphingomyelinase (aSMase) imipramine. The cholesterol sequestering agent nystatin also prevents cisplatin-induced CD95 clustering and decreases HT29 cell sensitivity to cisplatin-induced apoptosis and the synergy between cisplatin and anti-CD95 agonistic antibodies. CD95, together with the adaptor molecule Fas-associated death domain and procaspase-8, is redistributed into cholesterol- and sphingolipid-enriched cell fractions after cisplatin treatment, suggesting plasma membrane raft involvement. Interestingly, nystatin prevents the translocation of the aSMase to the extracellular surface of plasma membrane and the production of ceramide, suggesting that these early events require raft integrity. In addition, nystatin prevents cisplatin-induced transient increase in plasma membrane fluidity that could be required for CD95 translocation. Together, these results demonstrate that cisplatin activates aSMase and induces ceramide production, which triggers the redistribution of CD95 into the plasma membrane rafts. Such redistribution contributes to cell death and sensitizes tumor cells to CD95-mediated apoptosis.
- Published
- 2004
36. Increased immunogenicity of colon cancer cells by selective depletion of cytochrome C
- Author
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Eric Solary, Arlette Hammann, François Ghiringhelli, Elise Schmitt, Ahmed Hamaï, Noelia Casares, Sandeep Gurbuxani, Guido Kroemer, Monique Moutet, Carmen Garrido, Marie O. Pequignot, Nathalie Droin, Annie Fromentin, and Arnaud Parcellier
- Subjects
Cancer Research ,Programmed cell death ,Colorectal cancer ,Down-Regulation ,Apoptosis ,Transfection ,DNA, Antisense ,Mice ,Immune system ,In vivo ,medicine ,Animals ,Humans ,Etoposide ,chemistry.chemical_classification ,Reactive oxygen species ,biology ,Immunogenicity ,Cytochrome c ,Macrophages ,Cytochromes c ,medicine.disease ,Acquired immune system ,HCT116 Cells ,Staurosporine ,Rats ,Oncology ,Biochemistry ,chemistry ,Doxorubicin ,Colonic Neoplasms ,Cancer research ,biology.protein ,Cisplatin ,HT29 Cells - Abstract
We and others have previously reported in an in vivo rat colon cancer cell model that cell death precedes and is necessary for the development of a specific antitumoral immune response. To sensitize colon cancer cells to death, we depleted cytochrome c by stable transfection with an antisense construct. Cytochrome c depletion sensitizes human and rat colon cancer cells to a nonapoptotic, nonautophagic death induced by various stimuli. This increased sensitization to a necrosis-like cell death may be related to a decrease in cellular ATP levels and an increase in reactive oxygen species production caused by cytochrome c depletion. In vivo, depletion of cytochrome c decreases the tumorigenicity of colon cancer cells in syngeneic rats without influencing their growth in immune-deficient animals. Furthermore, decreased expression of cytochrome c in tumor cells facilitates in vivo “necrotic” cell death and the induction of a specific immune response. These results delineate a novel strategy to sensitize colon cancer cells to chemotherapy and to increase their immunogenicity in immuno-competent hosts.
- Published
- 2004
37. Influence of the nitric oxide donor glyceryl trinitrate on apoptotic pathways in human colon cancer cells
- Author
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Ali Bettaieb, Eric Solary, Arlette Hammann, Flore Renaud, Laurent Prevotat, Bernard Mignotte, Anne Millet, Jean-François Jeannin, Laboratoire de génétique et biologie cellulaire (LGBC), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), 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
Programmed cell death ,[SDV]Life Sciences [q-bio] ,CASP8 and FADD-Like Apoptosis Regulating Protein ,Caspase 3 ,Apoptosis ,Cytochrome c Group ,Flow cytometry ,03 medical and health sciences ,Nitroglycerin ,0302 clinical medicine ,medicine ,Tumor Cells, Cultured ,Humans ,Nitric Oxide Donors ,fas Receptor ,Caspase 10 ,Caspase ,ComputingMilieux_MISCELLANEOUS ,030304 developmental biology ,0303 health sciences ,Hepatology ,medicine.diagnostic_test ,biology ,Chemistry ,Receptor Aggregation ,Gastroenterology ,Intracellular Signaling Peptides and Proteins ,Fas receptor ,Mitochondria ,Enzyme Activation ,Biochemistry ,030220 oncology & carcinogenesis ,Caspases ,Death-inducing signaling complex ,Colonic Neoplasms ,biology.protein ,Cancer research ,Carrier Proteins - Abstract
Background & Aims: We have previously reported the role of nitric oxide in colon tumor regression in vivo. The present study was designed to explore the influence of an endogenous nitric oxide donor, glyceryl trinitrate (GTN), on cell death pathways in colon cancer cells. Methods: Human colon cancer cell lines were treated with the NO donor GTN. Apoptosis was identified by morphological criteria and the terminal deoxynucleotidyl transferase–mediated deoxyuridine (TUNEL) method. The mitochondrial transmembrane potential was studied by flow cytometry, cytochrome c release by Western blot, and caspase activation by combining fluorogenic peptide substrates, peptide inhibitors, and immunoblotting. Expression of death receptors was studied by flow cytometry and confocal microscopy. Results: GTN induces a dose- and time-dependent cell death by apoptosis in colon cancer cells. This cell death pathway involves the mitochondria and caspases, mainly caspase-1 and caspase-10. In contrast, caspase-3 activation is a late and limited event. Death receptors are not involved in GTN-mediated cell death, while GTN sensitizes tumor cells to Fas-ligand–induced apoptosis. This permissive effect correlates with an increased expression of Fas receptor and a decreased expression of several endogenous inhibitors of apoptosis (IAPs). Conclusions: Our results indicate that GTN (1) activates an unusual caspase cascade to induce apoptosis in colon cancer cells and (2) sensitizes these cells to Fas-mediated cell death by increasing the expression of Fas and decreasing the expression of several IAPs. GASTROENTEROLOGY 2002;123:235-246
- Published
- 2002
38. Involvement of caspase-2 long isoform in Fas-mediated cell death of human leukemic cells
- Author
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Arlette Hammann, Anne Wotawa, Olivier Sordet, Florence Bichat, Richard Bertrand, Nathalie Droin, Eric Solary, and Cédric Rébé
- Subjects
Immunology ,Caspase 2 ,CASP8 and FADD-Like Apoptosis Regulating Protein ,Apoptosis ,Cytochrome c Group ,Caspase 8 ,Transfection ,Biochemistry ,Fas ligand ,DNA, Antisense ,Tumor Cells, Cultured ,Humans ,Protein Isoforms ,FADD ,fas Receptor ,Leukemia ,biology ,Caspase 3 ,Intracellular Signaling Peptides and Proteins ,Cell Biology ,Hematology ,Fas receptor ,Caspase 9 ,Cell biology ,Caspases ,Death-inducing signaling complex ,biology.protein ,Carrier Proteins ,BH3 Interacting Domain Death Agonist Protein - Abstract
Engagement of the plasma membrane receptor Fas can induce apoptosis of leukemic cells. Signaling through Fas requires the formation of a death-inducing signaling complex (DISC) that involves the cytoplasmic domain of Fas, the adaptor molecule FADD/MORT-1, and procaspase-8. The present study investigated whether another caspase, known as procaspase-2L, played a role in Fas-mediated cell death. A series of human leukemic variant cells was derived by stable transfection with aCASP2L antisense construct (CASP2L/AS).Specific down-regulation of procaspase-2L decreased the sensitivity of these cells to apoptosis induced by an agonistic anti-Fas antibody (Ab, clone CH11), as determined by studying DNA fragmentation, chromatin condensation, and externalization of phosphatidylserine on the plasma membrane. In leukemic cells transfected with an empty vector, anti-Fas Ab treatment activated caspase-8, decreased the expression of the BH3 domain-only protein Bid, triggered the release of cytochrome c from the mitochondria to the cytosol, and activated caspase-3. All these events could not be observed when CASP2L/AS cells were similarly treated with anti-Fas Abs. CASP2L/AStransfection did not inhibit the formation of the DISC and no direct interaction between procaspase-2L and either Fas or FADD or procaspase-8 was identified. Down-regulation of procaspase-2L inhibited anti-Fas Ab–mediated cleavage of c-FLIP (FLICE-inhibitory protein), a protein that interferes with the formation of a functional DISC. These results suggest that the long isoform of caspase-2 plays a role in the Fas-mediated pathway to cell death by contributing to caspase-8 activation at the DISC level.
- Published
- 2001
39. Cutting edge: the tumor counterattack hypothesis revisited: colon cancer cells do not induce T cell apoptosis via the Fas (CD95, APO-1) pathway
- Author
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Bernard Bonnotte, Nathalie Favre-Felix, François Martin, Annie Fromentin, Arlette Hammann, and Eric Solary
- Subjects
Fas Ligand Protein ,T-Lymphocytes ,Immunology ,Apoptosis ,Biology ,Jurkat cells ,Fas ligand ,Jurkat Cells ,NK-92 ,Cancer stem cell ,Antigens, Neoplasm ,Immunology and Allergy ,Cytotoxic T cell ,Animals ,Humans ,RNA, Messenger ,fas Receptor ,Antigen-presenting cell ,Leukemia L1210 ,Lymphokine-activated killer cell ,Membrane Glycoproteins ,hemic and immune systems ,Coculture Techniques ,Growth Inhibitors ,Cell biology ,Cancer cell ,Antigens, Surface ,Colonic Neoplasms ,Cancer research ,Caco-2 Cells ,HT29 Cells ,Cell Division ,Signal Transduction - Abstract
The counterattack hypothesis, suggesting that cancer cells express Fas ligand (FasL) and are able to kill Fas-expressing tumor-infiltrating activated T cells, was supported by reports of the killing of Jurkat cells by FasL-expressing human colon cancer cell lines. Through the use of an improved cytotoxic assay in which soluble FasL and FasL-transfected KFL9 cells were used as positive controls, we show that none of seven human colon cancer cell lines induce apoptosis of two Fas-expressing target cell lines, Jurkat and L1210-Fas cells. Moreover, in coculture experiments, cancer cell monolayers do not inhibit the growth of Fas-expressing lymphoid cells. Although FasL mRNA and protein were detected in the extracts of the colon cancer cell lines, flow cytometry and confocal microscopy failed to detect the protein on the surface of tumor cells. These results suggest that the counterattack of tumor-infiltrating T lymphocytes by cancer cells may not account for immune tolerance toward tumor cells.
- Published
- 2000
40. HSP27 inhibits cytochrome c-dependent activation of procaspase-9
- Author
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Annie Fromentin, André Patrick Arrigo, Eric Solary, Jean-Marie Bruey, Carmen Garrido, and Arlette Hammann
- Subjects
endocrine system ,Programmed cell death ,animal structures ,Caspase 3 ,Apoptosis ,Cytochrome c Group ,urologic and male genital diseases ,Biochemistry ,Deoxyadenine Nucleotides ,Hsp27 ,Heat shock protein ,Genetics ,Humans ,Molecular Biology ,Heat-Shock Proteins ,Etoposide ,Caspase-9 ,Enzyme Precursors ,biology ,Chemistry ,Cytochrome c ,Cytochrome P450 reductase ,U937 Cells ,Antineoplastic Agents, Phytogenic ,Caspase Inhibitors ,Caspase 9 ,Cell biology ,Enzyme Activation ,Proto-Oncogene Proteins c-bcl-2 ,Drug Resistance, Neoplasm ,embryonic structures ,biology.protein ,Biotechnology - Abstract
We have previously shown that the small heat shock protein HSP27 inhibited apoptotic pathways triggered by a variety of stimuli in mammalian cells. The present study demonstrates that HSP27 overexpression decreases U937 human leukemic cell sensitivity to etoposide-induced cytotoxicity by preventing apoptosis. As observed for Bcl-2, HSP27 overexpression delays poly(ADP-ribose)polymerase cleavage and procaspase-3 activation. In contrast with Bcl-2, HSP27 overexpression does not prevent etoposide-induced cytochrome c release from the mitochondria. In a cell-free system, addition of cytochrome c and dATP to cytosolic extracts from untreated cells induces the proteolytic activation of procaspase-3 in both control and bcl-2-transfected U937 cells but fails to activate procaspase-3 in HSP27-overexpressing cells. Immunodepletion of HSP27 from cytosolic extracts increases cytochrome c/dATP-mediated activation of procaspase-3. Overexpression of HSP27 also prevents procaspase-9 activation. In the cell-free system, immunodepletion of HSP27 increases LEDH-AFC peptide cleavage activity triggered by cytochrome c/dATP treatment. We conclude that HSP27 inhibits etoposide-induced apoptosis by preventing cytochrome c and dATP-triggered activity of caspase-9, downstream of cytochrome c release.
- Published
- 1999
41. STAT-1-independent upregulation of FADD and procaspase-3 and -8 in cancer cells treated with cytotoxic drugs
- Author
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Eric Solary, Marie-Thérèse Dimanche-Boitrel, Olivier Micheau, and Arlette Hammann
- Subjects
Transcription, Genetic ,Fas-Associated Death Domain Protein ,Apoptosis ,Biochemistry ,Receptors, Tumor Necrosis Factor ,Tumor Cells, Cultured ,Cytotoxic T cell ,FADD ,bcl-2-Associated X Protein ,Caspase 8 ,Enzyme Precursors ,biology ,Chemistry ,Caspase 3 ,Caspase 2 ,Fas receptor ,Caspase 9 ,Cell biology ,Up-Regulation ,DNA-Binding Proteins ,Gene Expression Regulation, Neoplastic ,Isoenzymes ,STAT1 Transcription Factor ,Proto-Oncogene Proteins c-bcl-2 ,Caspases ,Colonic Neoplasms ,Dactinomycin ,Poly(ADP-ribose) Polymerases ,medicine.drug ,Programmed cell death ,Mitomycin ,Biophysics ,Antineoplastic Agents ,Downregulation and upregulation ,Proto-Oncogene Proteins ,medicine ,Humans ,RNA, Messenger ,fas Receptor ,Molecular Biology ,Adaptor Proteins, Signal Transducing ,Cisplatin ,Cell Biology ,Doxorubicin ,Cancer cell ,biology.protein ,Trans-Activators ,Carrier Proteins - Abstract
We have previously shown that treatment by anticancer drugs sensitized tumor cells to Fas (APO-1/CD95)-mediated cell death. The present study demonstrates that the cytotoxic drugs cisplatin, doxorubicin and mitomycin C induce the accumulation of the Fas receptor, the FADD adaptor molecule, the procaspases-8, -3 and -2L and the proapoptotic molecule Bax in several human colon cancer cells. This upregulation is also observed in U3A myeloblastoma cells that do not express STAT-1, a transcription factor involved in the constitutive expression of procaspases. We conclude that anticancer drugs sensitize tumor cells to Fas-mediated cell death by a STAT-1-independent upregulation of molecules involved in this apoptotic pathway.
- Published
- 1999
42. Fas ligand-independent, FADD-mediated activation of the Fas death pathway by anticancer drugs
- Author
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Arlette Hammann, Eric Solary, Marie-Thérèse Dimanche-Boitrel, and Olivier Micheau
- Subjects
Fatty Acid Desaturases ,Fas Ligand Protein ,Antineoplastic Agents ,Apoptosis ,Ligands ,Transfection ,Vinblastine ,Biochemistry ,Fas ligand ,Receptors, Tumor Necrosis Factor ,Jurkat Cells ,Tumor Cells, Cultured ,Cytotoxic T cell ,Humans ,FADD ,fas Receptor ,Molecular Biology ,Death domain ,Etoposide ,Plant Proteins ,Membrane Glycoproteins ,biology ,Arabidopsis Proteins ,Cell Biology ,U937 Cells ,Oligonucleotides, Antisense ,Fas receptor ,Flow Cytometry ,Cell biology ,Gene Expression Regulation ,Death-inducing signaling complex ,Antigens, Surface ,biology.protein ,Caspase 10 ,biological phenomena, cell phenomena, and immunity ,Cisplatin - Abstract
Trimerization of the Fas receptor (CD95, APO-1), a membrane bound protein, triggers cell death by apoptosis. The main death pathway activated by Fas receptor involves the adaptor protein FADD (for Fas-associated death domain) that connects Fas receptor to the caspase cascade. Anticancer drugs have been shown to enhance both Fas receptor and Fas ligand expression on tumor cells. The contribution of Fas ligand-Fas receptor interactions to the cytotoxic activity of these drugs remains controversial. Here, we show that neither the antagonistic anti-Fas antibody ZB4 nor the Fas-IgG molecule inhibit drug-induced apoptosis in three different cell lines. The expression of Fas ligand on the plasma membrane, which is identified in untreated U937 human leukemic cells but remains undetectable in untreated HT29 and HCT116 human colon cancer cell lines, is not modified by exposure to various cytotoxic agents. These drugs induce the clustering of Fas receptor, as observed by confocal laser scanning microscopy, and its interaction with FADD, as demonstrated by co-immunoprecipitation. Overexpression of FADD by stable transfection sensitizes tumor cells to drug-induced cell death and cytotoxicity, whereas down-regulation of FADD by transient transfection of an antisense construct decreases tumor cell sensitivity to drug-induced apoptosis. These results were confirmed by transient transfection of constructs encoding either a FADD dominant negative mutant or MC159 or E8 viral proteins that inhibit the FADD/caspase-8 pathway. These results suggest that drug-induced cell death involves the Fas/FADD pathway in a Fas ligand-independent fashion.
- Published
- 1999
43. Contribution of the cyclin-dependent kinase inhibitor p27KIP1 to the confluence-dependent resistance of HT29 human colon carcinoma cells
- Author
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Olivier Micheau, Monika Haugg, Bruno Chauffert, Arlette Hammann, Eric Solary, Marie-Thérèse Dimanche-Boitrel, Beatrice Eymin, Institut de recherche en santé, environnement et travail (Irset), Université d'Angers (UA)-Université de Rennes (UR)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Lipides - Nutrition - Cancer (U866) (LNC), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA), Institut National de la Santé et de la Recherche Médicale (INSERM), Groupe de Recherche sur le Cancer du Poumon (EA2021), Institut Albert Bonniot, Service d'oncologie, CHU Amiens-Picardie, Hématopoïèse normale et pathologique, Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Eymin, Beatrice, Université d'Angers (UA)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Gustave Roussy (IGR)-Université Paris-Sud - Paris 11 (UP11)
- Subjects
G2 Phase ,Cancer Research ,[SDV]Life Sciences [q-bio] ,Cyclin B ,Mitosis ,Apoptosis ,Cell Cycle Proteins ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,Transfection ,Cell Line ,HT29 Cells ,Cyclin-dependent kinase ,Cell Adhesion ,medicine ,Humans ,Point Mutation ,Cytotoxic T cell ,Enzyme Inhibitors ,[SDV.BC] Life Sciences [q-bio]/Cellular Biology ,Cisplatin ,Confluency ,biology ,Tumor Suppressor Proteins ,Cell Cycle ,Cell cycle ,Genes, p53 ,Cyclin-Dependent Kinases ,Recombinant Proteins ,3. Good health ,[SDV] Life Sciences [q-bio] ,Oncology ,Drug Resistance, Neoplasm ,Immunology ,biology.protein ,Cancer research ,Tumor Suppressor Protein p53 ,Microtubule-Associated Proteins ,Cyclin-Dependent Kinase Inhibitor p27 ,medicine.drug - Abstract
We have previously shown that growth of HT29 human colorectal cancer cells at confluence increased their resistance to the cytotoxic agent cisplatin. This study further explores the mechanisms of this resistance phenotype. DNA platination induced by cisplatin exposure is slightly reduced by confluence. However, at an equivalent DNA platination level, non-confluent cells accumulate in the G2/M phase of the cell cycle, demonstrate aberrant mitotic figures and die by apoptosis, while confluent cells progress slowly through the cell cycle, do not reach mitosis and are more resistant to drug-induced cell death. At a molecular level, cisplatin enhances cyclin B and p34cdc2 levels and histone H1 kinase activity in non-confluent, but not in confluent, cells. Furthermore, when HT29 cells reach confluence, expression of the cyclin-dependent kinase inhibitor p27Kip1 increases and cells accumulate in the G0/G1 phase of the cell cycle. Transfection-mediated over-expression of p27Kip1 in non-confluent HT29 cells decreases the cytotoxic activity of cisplatin as well as its ability to trigger apoptosis. Non-confluent HT29 cells over-expressing p27Kip1 are also more resistant to doxorubicin, etoposide and 5-fluorouracil. Our results suggest that p27Kip1 contributes to the confluence-dependent resistance phenotype. Int. J. Cancer 77:796–802, 1998. © 1998 Wiley-Liss, Inc.
- Published
- 1998
44. Sensitization of cancer cells treated with cytotoxic drugs to fas-mediated cytotoxicity
- Author
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François Martin, Olivier Micheau, Marie-Theérèse Dimanche-Boitrel, Eric Solary, and Arlette Hammann
- Subjects
Cancer Research ,Fas Ligand Protein ,Antineoplastic Agents ,Apoptosis ,Biology ,Fas ligand ,Mice ,Immune system ,Antigen ,medicine ,Tumor Cells, Cultured ,Cytotoxic T cell ,Animals ,Humans ,RNA, Messenger ,fas Receptor ,Membrane Glycoproteins ,Fas receptor ,Flow Cytometry ,Molecular biology ,Oncology ,Cancer cell ,Colonic Neoplasms ,Cisplatin ,Drug Screening Assays, Antitumor ,HT29 Cells ,Camptothecin ,medicine.drug - Abstract
Background The transmembrane receptor Fas, together with its protein-binding partner (Fas ligand), is a key regulator of programmed cell death (i.e., apoptosis). Fas and Fas ligand also influence the ability of cytotoxic T lymphocytes and natural killer cells to eliminate tumor cells. However, by inducing apoptosis in activated T cells, the Fas/Fas ligand system may protect some tumor cells from clearance by the immune system. Anticancer drugs enhance Fas ligand expression on the surface of Fas receptor-expressing leukemia cells, thus suggesting that apoptosis caused by these drugs may be mediated via the Fas/Fas ligand system. Purpose This study was conducted to further investigate the relationship between the modulation of Fas receptor gene and protein expression by treatment of cells with cytotoxic drugs and the immune clearance of tumor cells. Methods Fas expression on human HT29 colon carcinoma cells treated with a variety of anticancer drugs (cisplatin, doxorubicin, mitomycin C, fluorouracil, and camptothecin) was analyzed by use of quantitative flow cytometry. Human HCT8R and HCT116 colon carcinoma cells and human U937 leukemia cells were treated with cisplatin only and analyzed in the same way. Fas ligand messenger RNA and protein levels were studied by use of a reverse transcription-polymerase chain reaction assay and by flow cytometry. Fas gene expression and messenger RNA levels in cisplatin-treated HT29 cells were characterized by use of in vitro nuclear run-on and northern blot hybridization assays. The cytotoxic activities of agonistic anti-Fas antibodies, Fas ligand, and allogeneic peripheral blood leukocytes, in the absence or presence of Fas-blocking monoclonal antibodies, against tumor cells were assessed by methylene blue staining and chromium-51 release assays. Results Clinically relevant concentrations of cisplatin, doxorubicin, mitomycin C, fluorouracil, or camptothecin enhanced Fas receptor expression on the plasma membrane of HT29 cells. Cisplatin-mediated increases in Fas expression were confirmed in HCT8R, HCT116, and U937 cells. The enhancement of Fas protein expression was associated with an increased sensitivity of cisplatin-treated tumor cells to agonistic anti-Fas antibodies, to soluble Fas ligand, and to allogeneic peripheral blood leukocyte-mediated cytotoxicity. Each of these effects was blocked by co-treatment of the cells with antagonistic anti-Fas antibody. Conclusion and implications In addition to their direct cytotoxic effects, chemotherapeutic drugs sensitize tumor cells to Fas-mediated cytotoxicity and Fas-dependent immune clearance. On the basis of these findings, new strategies might be developed to improve the efficacy of these drugs.
- Published
- 1997
45. Abnormal basement membrane in tumors induced by rat colon cancer cells
- Author
-
Arlette Hammann, Monique Martin, Philippe Pujuguet, and François Martin
- Subjects
Pathology ,medicine.medical_specialty ,Colorectal cancer ,Colon ,Perlecan ,Biology ,Adenocarcinoma ,Basement Membrane ,Extracellular matrix ,Type IV collagen ,Laminin ,Reference Values ,medicine ,Carcinoma ,Animals ,Basement membrane ,Hepatology ,Staining and Labeling ,Gastroenterology ,Muscle, Smooth ,Fibroblasts ,medicine.disease ,Immunohistochemistry ,Extracellular Matrix ,Rats ,medicine.anatomical_structure ,Colonic Neoplasms ,biology.protein ,Collagen ,Myofibroblast ,Neoplasm Transplantation - Abstract
Background/Aims: Colonic mucosa basement membrane results from a cooperation between epithelial cells and pericryptal fibroblasts characterized as myofibroblasts. This cooperation may be abnormal in colorectal carcinoma resulting in basement membrane alteration. Methods: Basement membrane composition and myofibroblast distribution were studied in normal rat colon and two colon carcinoma models by immunohistochemistry. Colon cancer cells and tumor-associated myofibroblasts were also studied for their capacity to deposit three basement membrane components (laminin, heparan sulfate proteoglycan, and type IV collagen) in vitro. Results: A continuous, type IV collagen-containing basement membrane, such as that observed in normal colon, was found only in the most differentiated tumor model and was restricted to the areas in which myofibroblasts were closely apposed to carcinoma cells. In other areas of this tumor and in the poorly differentiated tumor model, myofibroblasts dissociated from the epithelial cells and the basement membrane was devoid of type IV collagen. In vitro, carcinoma cells deposited laminin and heparan sulfate proteoglycan but not type IV collagen. Tumor-associated myofibroblasts deposited type IV collagen only in the presence of tumor cell extracellular matrix or laminin coating. Conclusions: The colon cancer basement membrane defect in type IV collagen may result from a physical disruption in the association between epithelial cancer cells and myofibroblasts.
- Published
- 1994
46. In vivo and in vitro invasiveness of a rat colon-cancer cell line maintaining E-cadherin expression: an enhancing role of tumor-associated myofibroblasts
- Author
-
Arlette Hammann, François Martin, Monique Martin, Luc Vakaet, Bruno Chauffert, Frans van Roy, Marc Mareel, Marie Thearèse Dimanche‐Boitrel, and Philippe Pujuguet
- Subjects
Cancer Research ,Pathology ,medicine.medical_specialty ,Cell ,Biology ,Metastasis ,Extracellular matrix ,In vivo ,medicine ,Tumor Cells, Cultured ,Animals ,Neoplasm Invasiveness ,Cadherin ,Myocardium ,Rats, Inbred Strains ,Fibroblasts ,medicine.disease ,Cadherins ,Immunohistochemistry ,In vitro ,Cell biology ,Rats ,medicine.anatomical_structure ,Oncology ,Cell culture ,Colonic Neoplasms ,Collagenase ,Chickens ,medicine.drug - Abstract
In various cell systems, an inverse relationship was found between expression of E-cadherin, a molecule involved in the Ca2+-dependent homophylic cell-to-cell attachment of epithelial cells, and the capacity to invade extracellular matrix gels or normal tissues in vitro., DHD/K12/TRb (PROb) cells, maintained as a cell line derived from a rat colon carcinoma, homogeneously expressed. in vitro immunoreactive E-cadherin, which was functional as shown in cell dissociation-reassociation assays. PROb cells were found to be non-invasive in 3 different assays in vitro., However, tumors resulting from a s.c. injection of PROb cells into syngeneic BD-IX rats were invasive, although PROb cells maintained E-cadherin expression in the tumors. Cells from a freshly dissociated PROb tumor showed, not only PROb cells but also tumor-associated myofibrobfasts and were able to cross a Matrigel-coated filter. PROb tumors were indeed infiltrated by numerous myofibroblasts, mainly located at the invasive edge of the tumor. Cells from an established culture of tumor-infiltrating myofibroblasts were able to confer upon PROb cells invasiveness through Matrigel-coated filter or into chick-heart fragments. PROb cells maintained their capacity to express E-cadherin after myofibroblast-enhanced Matrigel invasion. Tumor-associated myofibroblasts, but not PROb cells, secreted a 72-kDa collagenase that could play a role in tumor-cell invasion. These results strongly suggest that cells from the tumor stroma, and more specifically myofibroblasts, may be involved in the invasiveness of epithelial tumor cells in vivo, even when E-cadherin expression prevents tumor-cell invasiveness in different in vitro assays.
- Published
- 1994
47. Tif1gamma Is Essential for Macrophage Differentiation
- Author
-
Laurent Delva, Jean-Noël Bastie, Eric Solary, Marie-Lorraine Chretien, Nathalie Droin, Romain Aucagne, and Arlette Hammann
- Subjects
Tumor suppressor gene ,Monocyte ,medicine.medical_treatment ,Immunology ,CD34 ,Cell Biology ,Hematology ,Biology ,Biochemistry ,Cell biology ,Haematopoiesis ,medicine.anatomical_structure ,Cytokine ,medicine ,Bone marrow ,Stem cell ,Progenitor cell - Abstract
Abstract 2370 TIF1gamma (or TRIM33) is an ubiquitous nuclear protein that belongs to the transcriptional intermediary factor 1 family. Human and mouse TIF1gamma are closely related to zebrafish moonshine (mon), a gene whose mutations disrupt embryonic and adult hematopoiesis with severe red blood cell aplasia. Targeted deletion of Tif1gamma is embryonic lethal in mice. In zebrafish and human CD34+ cells, TIF1gamma functionally links positive elongation factors such as p-TEFb and FACT to blood specific transcription complexes (e.g. the SCL/TAL1 complex) to regulate elongation of genes by antagonizing Pol II pausing. TIF1gamma also affects the human hematopoietic progenitor cell response to the cytokines of the transforming growth factor-beta superfamily through various mechanisms. Recently, we showed that the loss of Tif1gamma in mouse hematopoietic stem cells (cFES-Cre-Tif1gamma) favors the expansion of the granulo-monocytic progenitor compartment. The gene deletion induces the age-dependent appearance of a cell-autonomous myeloproliferative disorder with myelodysplastic features, monocytosis, and hepatosplenomegaly that recapitulates essential features of human chronic myelomonocytic leukemia (CMML). Interestingly, TIF1gamma is almost undetectable in leukemic cells of 35% of patients with CMML. This down-regulation is related to the hyper-methylation of CpG sequences in the gene promoter. Our results demonstrated that TIF1gamma is an epigenetically regulated tumor suppressor gene in hematopoietic cells. In addition, an altered production of peritoneal macrophages was observed in our mouse model. These macrophages did not adhere to the plastic and were morphologically abnormal in vitro. In bone marrow and in Lin- progenitor cells, Tif1gamma deletion leads to a significant decrease of cfms (Csf-1r) expression, required for the differentiation, proliferation, and survival of monocytic phagocytes. We also identified in CMML patients the association between low levels of TIF1gamma and cFMS (Aucagne et al., J. Clin. Invest., 121, 2361–2370, 2011). To gain insight into the possible mechanism accounting for diminished accumulation of macrophages, we examined the expression of c-Fms. We show that level of its expression is reduced significantly in blood monocytes isolated from Tif1gamma-deleted mice. When Tif1gamma-deleted sorted myeloid cells were induced to differentiate into macrophages in presence of CSF-1, a delayed production of few abnormal large macrophages was observed. Apoptosis was associated with this alteration of differentiation. This phenomenon was also characterized in young mice not developing the disease yet. The morphological abnormalities were correlated with very important alterations of specific macrophage differentiation markers. Expression of specific transcription factors involved in macrophage differentiation was deeply deregulated. Moreover, macrophage function such as migration, cytokine or chemokine secretion in response to LPS was altered. Likewise, in vitro differentiation of monocytes into dendritic cells was also abnormal. Altogether, our results suggest that monocyte plasticity is at least partially orchestrated by Tif1gamma. Disclosures: No relevant conflicts of interest to declare.
- Published
- 2011
48. Surface phenotype and functions of tumor-infiltrating dendritic cells: CD8 expression by a cell subpopulation
- Author
-
François Martin, Arlette Hammann, Pascal Chaux, and Monique Martin
- Subjects
Pathology ,medicine.medical_specialty ,CD8 Antigens ,Immunology ,Antigen presentation ,Antigen-Presenting Cells ,Biology ,Major histocompatibility complex ,Cell morphology ,Lymphocytes, Tumor-Infiltrating ,Antigen ,Antigens, Neoplasm ,medicine ,Immunology and Allergy ,Animals ,Antibodies, Monoclonal ,Rats, Inbred Strains ,Dendritic cell ,Dendritic Cells ,Neoplasms, Experimental ,Molecular biology ,Tumor antigen ,Rats ,Phenotype ,Cancer cell ,biology.protein ,CD8 - Abstract
Although the function and significance of tumor-infiltrating dendritic cells (TIDC) in the immune response to tumor have never been clearly demonstrated, their location suggests that they play a critical role in the presentation of tumor antigen to specific T cells. We studied the morphological and functional characteristics of interstitial dendritic cells (DC) located inside tumors obtained by injection of cancer cells into syngeneic rats. Single and double immunostaining of tumor sections revealed a dense network of cells which expressed class II major histocompatibility complex (MHC II) molecules. Cell morphology and surface markers were characteristic of DC populations in other tissues. These DC were in close contact with tumor cells and increased in number as the tumor grew larger. Unexpectedly, a subpopulation of morphologically characteristic TIDC expressed both CD8 and MHC II molecules. TIDC were purified from tumors by gradient centrifugation and immunobeads and characterized by morphology, ultrastructural study and surface markers studied by flow cytometry. TIDC were negative for the CD5 molecule (a pan T cell marker), and were not labeled with 3.2.3 monoclonal antibody (mAb) (an NK cell marker) or with Ki-M2R mAb (a macrophage marker). A subpopulation of TIDC expressed the CD8 molecule, confirming the in situ results. TIDC expressed high levels of class I and class II MHC molecules and the adhesion molecule ICAM-1. This expression is compatible with effective antigen presenting function. Purified TIDC triggered rapid and high levels of proliferation of tumor-immune T cells in vitro, demonstrating the potential of these cells to constitutively process and present tumor-associated antigens.
- Published
- 1993
49. MYST3/NCOA2-Induced Acute Myeloid Leukemia in Transgenic Fish
- Author
-
Arlette Hammann, Eric Solary, Julia Zhuravleva, Laurent Martin, Laurent Delva, Jean-Noël Bastie, and Jérôme Paggetti
- Subjects
Myeloid ,biology ,Immunology ,RUNX1T1 ,Myeloid leukemia ,Cell Biology ,Hematology ,biology.organism_classification ,medicine.disease ,Biochemistry ,Molecular biology ,Fusion gene ,ETV6 ,Leukemia ,chemistry.chemical_compound ,medicine.anatomical_structure ,RUNX1 ,chemistry ,hemic and lymphatic diseases ,medicine ,Zebrafish - Abstract
The MYST3/NCOA2 (MOZ/TIF2) fusion gene generated by the inv(8)(p11q13) chromosomal abnormality was described in a specific subgroup of acute myeloid leukemias (AML) that represents less than 5% of AML4/5. This abnormality fuses MYST3 (MOZ), a member of the MYST family of histone acetyl-transferases (HAT) to NCOA2 (TIF2), a member of the p160 HAT family. The transforming properties of MYST3/NCOA2 were demonstrated in mouse committed myeloid progenitors in vitro and in vivo. Hematopoiesis is very similar in zebrafish and in higher vertebrates. Homologues of a large number of genes involved in mammalian myelopoiesis were identified in this animal model. We have recently shown that ncoa2 (tif2) played a role in zebrafish primitive hematopoiesis. This animal also represents a model for investigating leukemogenesis. Transgenic expression of rag2-EGFP-mMyc or rag2-ICN1-EGFP induces a T-cell acute lymphoblastic leukemia (ALL) whereas transgenic expression of the ETV6/RUNX1 fusion gene induces a B-cell type ALL. We generated a transgenic zebrafish in which the MYST3/NCOA2 fusion gene was expressed under control of the spi1 (pu.1) promoter. An AML developed in two of 180 MYST3/NCOA2-EGFP-expressing embryos, 14 and 26 months after injection of the fusion gene in a one cell embryo, respectively. This leukemia was characterized by an extensive invasion of kidneys by myeloid blast cells. This model, which is the first zebrafish model of acute myeloid leukemia, demonstrates the oncogenic potency of MYST3/NCOA2 fusion gene.
- Published
- 2008
50. An immunohistological study of cells infiltrating progressive and regressive tumors induced by two variant subpopulations of a rat colon cancer cell line
- Author
-
Arlette Hammann, Anne Caignard, H. Pelletier, Monique Martin, and François Martin
- Subjects
Cancer Research ,Pathology ,medicine.medical_specialty ,Histocytochemistry ,Antibodies, Monoclonal ,Histology ,T lymphocyte ,Biology ,Cell Line ,Rats ,Immunoenzyme Techniques ,Kinetics ,Oncology ,Cell culture ,Tumor progression ,Monoclonal ,Colonic Neoplasms ,medicine ,Immunohistochemistry ,Macrophage ,Cytotoxic T cell ,Animals - Abstract
In order to understand the mechanisms leading up to progression or regression, tumors resulting from the s.c. inoculation of progressive or regressive variants of a cell culture established from a chemically-induced rat colonic carcinoma were subjected to sequential histological study. As immunological factors have been previously described in this system of progressive or regressive tumors, special interest was given to inflammatory cells, T and B lymphocytes and macrophages, located inside and outside the tumor. Immunohistological methods using monoclonal or polyclonal antibodies and enzyme histology were performed to identify different populations of infiltrative cells. In both variants of tumors an accumulation of these cells were seen at the periphery of the tumor, surrounding the nodules. In contrast, very few inflammatory cells, macrophages or T lymphocytes were seen inside the clumps of tumor cells where cytolytic cells could have a contact-dependent tumoricidal effect. Only small differences were found between progressive and regressive tumors in the density of the various populations of T helper, T cytotoxic/suppressor, B lymphocytes or macrophages inside or around the tumor nodules. On the other hand, progressive and regressive tumors clearly differ in the relationship between tumor cells and the fibroblastic reaction they induce. Regressive tumors were rapidly encircled by a fibroblastic reaction isolating them from the peripheral tissues. The fibroblastic reaction was less dense around the progressive tumor cells which were able to migrate and invade the periphery. This suggests that immunological factors leading to tumor progression or regression could act indirectly through a control of the fibroblastic reaction, rather than through a direct cytotoxic effect on the tumor cells.
- Published
- 1987
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