102 results on '"Barkats M"'
Search Results
2. Gene therapy approaches for familial ALS
- Author
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Cappella, Marisa, Cohen-Tannoudji, Mathilde, Marais, Thibaut, Astord, Stéphanie, Besse, Aurore, Giroux, Benoit, Lefebvre, Cynthia, Bohl, Delphine, Barkats, M., Biferi, Maria, Centre de recherche en Myologie – U974 SU-INSERM, Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Institut de Myologie, Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Association française contre les myopathies (AFM-Téléthon)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Institut du Cerveau = Paris Brain Institute (ICM), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], and Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
[SDV]Life Sciences [q-bio] - Published
- 2018
3. AAV9-mediated SMN expression restricted to theCNS does not rescue SMA mice
- Author
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Besse, Aurore, Roda, Mariane, Astord, Stéphanie, Marais, Thibaut, Biferi, Maria, Barkats, M., Centre de recherche en Myologie – U974 SU-INSERM, Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Institut de Myologie, and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Association française contre les myopathies (AFM-Téléthon)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
[SDV]Life Sciences [q-bio] - Published
- 2017
4. AAV9-mediated SMN expression restricted to the CNS does not rescue SMA mice
- Author
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Besse, A., primary, Roda, M., additional, Astord, S., additional, Marais, T., additional, Biferi, M., additional, and Barkats, M., additional
- Published
- 2017
- Full Text
- View/download PDF
5. Whole-body rescue of Pompe disease with AAV liver delivery of engineered secretable GAA transgenes
- Author
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Colella, P., primary, Puzzo, F., additional, Biferi, M., additional, Bali, D., additional, Paulk, N., additional, Vidal, P., additional, Collaud, F., additional, Simon-Sola, M., additional, Charles, S., additional, Hardet, R., additional, Leborgne, C., additional, Sellier, P., additional, van Wittenberghe, L., additional, Boisgerault, F., additional, Barkats, M., additional, Laforêt, P., additional, Kay, M., additional, Koeberl, D., additional, Ronzitti, G., additional, and Mingozzi, F., additional
- Published
- 2017
- Full Text
- View/download PDF
6. A new AAV10-mediated gene therapy for SOD1 -linked ALS
- Author
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Biferi, M., primary, Cohen-Tannoudji, M., additional, Cappelletto, A., additional, Giroux, B., additional, Roda, M., additional, Astord, S., additional, Marais, T., additional, Ferry, A., additional, Voit, T., additional, and Barkats, M., additional
- Published
- 2017
- Full Text
- View/download PDF
7. Role of muscle satellite cells in Spinal muscular atrophy physiopathology
- Author
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Mecca, J., primary, Astord, S., additional, Marais, T., additional, Relaix, F., additional, Didier, N., additional, and Barkats, M., additional
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- 2017
- Full Text
- View/download PDF
8. Implication of the SMN complex in the biogenesis and steady state level of the Signal Recognition Particle
- Author
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Piazzon, N., Schlotter, F., Lefebvre, S., Dodre, M., Mereau, A., Soret, J., Besse, A., Barkats, M., Bordonne, R., Branlant, C., Massenet, S., Institut de Génétique Moléculaire de Montpellier (IGMM), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Institut Jacques Monod ( IJM ), Université Paris Diderot - Paris 7 ( UPD7 ) -Centre National de la Recherche Scientifique ( CNRS ), Institut de Génétique et Développement de Rennes ( IGDR ), Université de Rennes 1 ( UR1 ), Université de Rennes ( UNIV-RENNES ) -Université de Rennes ( UNIV-RENNES ) -IFR140-Centre National de la Recherche Scientifique ( CNRS ), Institut de Génétique Moléculaire de Montpellier ( IGMM ), Université de Montpellier ( UM ) -Centre National de la Recherche Scientifique ( CNRS ), Génétique moléculaire de la neurotransmission et des processus neurodégénératifs ( LGMNPN ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Centre National de la Recherche Scientifique ( CNRS ), ARN-RNP, structure-fonction-maturation ( AREMS ), Université Henri Poincaré - Nancy 1 ( UHP ) -Centre National de la Recherche Scientifique ( CNRS ), ARN-RNP, structure-fonction-maturation (AREMS), Université Henri Poincaré - Nancy 1 (UHP)-Centre National de la Recherche Scientifique (CNRS), Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Institut de Génétique et Développement de Rennes (IGDR), Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Centre de recherche en myologie, Université Pierre et Marie Curie - Paris 6 (UPMC)-Association française contre les myopathies (AFM-Téléthon)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Génétique moléculaire de la neurotransmission et des processus neurodégénératifs (LGMNPN), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), Université de Paris (UP)-Centre National de la Recherche Scientifique (CNRS), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Jonchère, Laurent, Laboratoire d’Océanologie et de Géosciences (LOG) - UMR 8187 (LOG), Institut national des sciences de l'Univers (INSU - CNRS)-Université de Lille-Université du Littoral Côte d'Opale (ULCO)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Physique Théorique d'Orsay [Orsay] (LPT), Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11), Institut National de la Santé et de la Recherche Médicale (INSERM), Ingénierie Moléculaire et Physiopathologie Articulaire (IMoPA), Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Henri Poincaré - Nancy 1 (UHP), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS), and Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP)
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Cytoplasm ,[SDV]Life Sciences [q-bio] ,animal diseases ,Molecular Sequence Data ,environment and public health ,Antibodies ,Muscular Atrophy, Spinal ,Mice ,Xenopus laevis ,[SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN] ,RNA, Small Nuclear ,RNA, Small Cytoplasmic ,Schizosaccharomyces ,Animals ,Humans ,Alleles ,ComputingMilieux_MISCELLANEOUS ,[SDV.GEN]Life Sciences [q-bio]/Genetics ,Base Sequence ,SMN Complex Proteins ,[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology ,nervous system diseases ,Spinal Cord ,nervous system ,Mutation ,[SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN] ,RNA ,[ SDV.GEN ] Life Sciences [q-bio]/Genetics ,Signal Recognition Particle ,HeLa Cells - Abstract
International audience; Spinal muscular atrophy is a severe motor neuron disease caused by reduced levels of the ubiquitous Survival of MotoNeurons (SMN) protein. SMN is part of a complex that is essential for spliceosomal UsnRNP biogenesis. Signal recognition particle (SRP) is a ribonucleoprotein particle crucial for co-translational targeting of secretory and membrane proteins to the endoplasmic reticulum. SRP biogenesis is a nucleo-cytoplasmic multistep process in which the protein components, except SRP54, assemble with 7S RNA in the nucleolus. Then, SRP54 is incorporated after export of the pre-particle into the cytoplasm. The assembly factors necessary for SRP biogenesis remain to be identified. Here, we show that 7S RNA binds to purified SMN complexes in vitro and that SMN complexes associate with SRP in cellular extracts. We identified the RNA determinants required. Moreover, we report a specific reduction of 7S RNA levels in the spinal cord of SMN-deficient mice, and in a Schizosaccharomyces pombe strain carrying a temperature-degron allele of SMN. Additionally, microinjected antibodies directed against SMN or Gemin2 interfere with the association of SRP54 with 7S RNA in Xenopus laevis oocytes. Our data show that reduced levels of the SMN protein lead to defect in SRP steady-state level and describe the SMN complex as the first identified cellular factor required for SRP biogenesis.
- Published
- 2013
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9. rAAV9-mediated gene transfer in the spinal cord of a feline model of motor neuron degeneration
- Author
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Bucher, T., Maquigneau, M., Dubreil, Laurence, Briot-Nivard, D., Wakeling, E., Fyfe, J., Costiou, P., Moullier, P., Barkats, M., Colle, Marie-Anne, Joussemet, B., Laboratoire de Thérapie Génique Translationnelle des Maladies Génétiques (Inserm UMR 1089), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN), Physiopathologie Animale et bioThérapie du muscle et du système nerveux (PAnTher), Institut National de la Recherche Agronomique (INRA)-Ecole Nationale Vétérinaire, Agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS), Department of Microbiology and Molecular Genetics, Michigan State University [East Lansing], Michigan State University System-Michigan State University System, Ecole Nationale Vétérinaire, Agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS), University of Florida [Gainesville] (UF), Institut National de la Santé et de la Recherche Médicale (INSERM), Université Pierre et Marie Curie - Paris 6 (UPMC), Centre National de la Recherche Scientifique (CNRS), Institut de Myologie, Institut National de la Recherche Agronomique (INRA)-École nationale vétérinaire, agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS), École nationale vétérinaire, agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Association française contre les myopathies (AFM-Téléthon)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
[SDV]Life Sciences [q-bio] ,ComputingMilieux_MISCELLANEOUS - Abstract
National audience
- Published
- 2012
10. Modeling and pathophysiological analysis of the ubiquilin-2-linked amyotrophic lateral sclerosis (ALS) with AAV vectors
- Author
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Bos, C., primary, Biferi, M., additional, Cohen-Tannoudji, M., additional, Roda, M., additional, and Barkats, M., additional
- Published
- 2015
- Full Text
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11. AAV1-, AAV2- and AAV5-Mediated human alpha-Iduronidase Gene Transfer In The Brain of Nonhuman Primate: Vector Diffusion and Bio Distribution
- Author
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Ciron, Carine, Cressant, A, Roux, F, Raoul, Solène, Cherel, Yan, Hantraye, P, Déglon, Nicole, Schwartz, Bertrand, Barkats, M, Heard, Jm, Tardieu, M, Moullier, P, Colle, Marie-Anne, Inconnu, Développement et Pathologie du Tissu Musculaire (DPTM), Institut National de la Recherche Agronomique (INRA)-Ecole Nationale Vétérinaire de Nantes, Plateforme d'Infectiologie Expérimentale (PFIE), Institut National de la Recherche Agronomique (INRA), and ProdInra, Migration
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[SDV] Life Sciences [q-bio] ,[SDV]Life Sciences [q-bio] ,ComputingMilieux_MISCELLANEOUS - Abstract
International audience
- Published
- 2009
12. TH.O.16 - A new AAV10-mediated gene therapy for SOD1-linked ALS
- Author
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Biferi, M., Cohen-Tannoudji, M., Cappelletto, A., Giroux, B., Roda, M., Astord, S., Marais, T., Ferry, A., Voit, T., and Barkats, M.
- Published
- 2017
- Full Text
- View/download PDF
13. TH.O.17 - Whole-body rescue of Pompe disease with AAV liver delivery of engineered secretable GAA transgenes
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Colella, P., Puzzo, F., Biferi, M., Bali, D., Paulk, N., Vidal, P., Collaud, F., Simon-Sola, M., Charles, S., Hardet, R., Leborgne, C., Sellier, P., van Wittenberghe, L., Boisgerault, F., Barkats, M., Laforêt, P., Kay, M., Koeberl, D., Ronzitti, G., and Mingozzi, F.
- Published
- 2017
- Full Text
- View/download PDF
14. P.132 - AAV9-mediated SMN expression restricted to the CNS does not rescue SMA mice
- Author
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Besse, A., Roda, M., Astord, S., Marais, T., Biferi, M., and Barkats, M.
- Published
- 2017
- Full Text
- View/download PDF
15. P.131 - Role of muscle satellite cells in Spinal muscular atrophy physiopathology
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Mecca, J., Astord, S., Marais, T., Relaix, F., Didier, N., and Barkats, M.
- Published
- 2017
- Full Text
- View/download PDF
16. Widespread gene delivery to motor neurons using peripheral injection of AAV vectors
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Joussemet, Béatrice, Riviere, Coraline, Marais, T, Douar, Am, Colle, Marie-Anne, Moullier, P, Barkats, M, Inconnu, and ProdInra, Migration
- Subjects
[SDV] Life Sciences [q-bio] ,[SDV]Life Sciences [q-bio] - Published
- 2007
17. AAV-mediated gene transfer in the central nervous system of a feline model of spinal muscular atrophy
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Duque, S., Colle, M.-A., Moullier, P., Barkats, M., and ProdInra, Migration
- Subjects
[SDV] Life Sciences [q-bio] ,[SDV]Life Sciences [q-bio] - Published
- 2007
18. G.O.20
- Author
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Biferi, M.G., primary, Bos, C., additional, Tanguy, Y., additional, Alonso-Martin, S., additional, Artegiani, B., additional, Roda, M., additional, Cohen-Tannoudji, M., additional, Ferry, A., additional, Camerini, S., additional, Crescenzi, M., additional, Calegari, F., additional, Relaix, F., additional, and Barkats, M., additional
- Published
- 2014
- Full Text
- View/download PDF
19. Implication of the SMN complex in the biogenesis and steady state level of the Signal Recognition Particle
- Author
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Piazzon, N., primary, Schlotter, F., additional, Lefebvre, S., additional, Dodre, M., additional, Mereau, A., additional, Soret, J., additional, Besse, A., additional, Barkats, M., additional, Bordonne, R., additional, Branlant, C., additional, and Massenet, S., additional
- Published
- 2012
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20. P3.10 Intramuscular AAV9 administration enables transgene delivery to motor neurons in the whole spinal cord: Therapeutic application in a SMA mouse model
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Benkhelifa-Ziyyat, S., primary, Besse, A., additional, Duqué, S., additional, Carcenac, R., additional, Marais, T., additional, Astord, S., additional, Roda, M., additional, and Barkats, M., additional
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- 2011
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21. G.P.133 - Modeling and pathophysiological analysis of the ubiquilin-2-linked amyotrophic lateral sclerosis (ALS) with AAV vectors
- Author
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Bos, C., Biferi, M., Cohen-Tannoudji, M., Roda, M., and Barkats, M.
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- 2015
- Full Text
- View/download PDF
22. O.18 Intravenous injection of SMN1-expressing self-complementary AAV9 rescues severe type I SMA mice
- Author
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Dominguez, E., primary, Chatauret, N., additional, Marais, T., additional, Duque, S., additional, Carcenac, R., additional, Astord, S., additional, de Moura, A. Pereira, additional, Voit, T., additional, and Barkats, M., additional
- Published
- 2010
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23. G.O.4 The NF-kappaB activator PLEKHG5 gene is mutated in a form of autosomal recessive lower motor neuron disease with childhood onset
- Author
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Maystadt, I., primary, Rezsöhazy, R., additional, Barkats, M., additional, Duque, S., additional, Vannuffel, P., additional, Najimi, M., additional, Munnich, A., additional, Viollet, L., additional, and Verellen-Dumoulin, C., additional
- Published
- 2007
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24. G.O.20: AAV-mediated overexpression of Ubiquilin2 mimics ALS and ALS with dementia in naive mice
- Author
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Biferi, M.G., Bos, C., Tanguy, Y., Alonso-Martin, S., Artegiani, B., Roda, M., Cohen-Tannoudji, M., Ferry, A., Camerini, S., Crescenzi, M., Calegari, F., Relaix, F., and Barkats, M.
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- 2014
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25. Adenovirus in the brain: recent advances of gene therapy for neurodegenerative diseases
- Author
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Barkats, M., primary, Bilang-Bleuel, A., additional, Buc-Caron, M.H., additional, Castel-Barthe, M.N., additional, Corti, O., additional, Finiels, F., additional, Horellou, P., additional, Revah, F., additional, Sabate, O., additional, and Mallet, J., additional
- Published
- 1998
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26. Effects of Ginkgo biloba extract (EGb 761) on learning and possible actions on aging
- Author
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Cohen-Salmon, Ch, primary, Venault, P, additional, Martin, B, additional, Raffalli-Sébille, M-J, additional, Barkats, M, additional, Clostre, F, additional, Pardon, M-C, additional, Christen, Y, additional, and Chapouthier, G, additional
- Published
- 1997
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27. Intrastriatal grafts of embryonic mesencephalic rat neurons genetically modified using an adenovirus encoding human Cu/Zn superoxide dismutase
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Barkats, M, primary, Nakao, N, additional, Grasbon-Frodl, E.M, additional, Bilang-Bleuel, A, additional, Revah, F, additional, Mallet, J, additional, and Brundin, P, additional
- Published
- 1997
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28. Age-related morphological changes in the hippocampus in two mouse strains
- Author
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Barkats, M., primary, Bertholet, J.-Y., additional, and Cohen-Salmon, C., additional
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- 1996
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29. Hippocampal mossy fiber changes in mice transgenic for the human copper-zinc superoxide dismutase gene
- Author
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Barkats, M., primary, Bertholet, J.-Y., additional, Venault, P., additional, Ceballos-Picot, I., additional, Nicole, A., additional, Phillips, J., additional, Moutier, R., additional, Roubertoux, P., additional, Sinet, P.-M., additional, and Cohen-Salmon, C., additional
- Published
- 1993
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30. Neuronal transfer of the human Cu/Zn superoxide dismutase gene increases the resistance of dopaminergic neurons to 6-hydroxydopamine.
- Author
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Barkats, M., Millecamps, S., Bilang-Bleuel, A., and Mallet, J.
- Subjects
- *
SUPEROXIDE dismutase , *SUBSTANTIA nigra , *DOPAMINERGIC neurons , *GENETIC transformation - Abstract
Several mechanisms are thought to be involved in the progressive decline in neurons of the substantia nigra pars compacta (SNpc) that leads to Parkinson's disease (PD). Neurotoxin 6-hydroxydopamine (6-OHDA), which induces parkinsonian symptoms in experimental animals, is thought to be formed endogenously in patients with PD through dopamine (DA) oxidation and may cause dopaminergic cell death via a free radical mechanism. We therefore investigated protection against 6-OHDA by inhibiting oxidative stress using a gene transfer strategy. We overexpressed the antioxidative Cu/Zn-superoxide dismutase (SOD1) enzyme in primary culture dopaminergic cells by infection with an adenovirus carrying the human SOD1 gene (Ad-hSOD1). Survival of the dopaminergic cells exposed to 6-OHDA was 50% higher among the SOD1-producing cells than the cells infected with control adenoviruses. In contrast, no significant increased survival of (6-OHDA)-treated dopaminergic cells was observed when they were infected with an adenovirus expressing the H[sub 2]O[sub 2]-scavenging glutathione peroxidase (GPx) enzyme. These results underline the major contribution of superoxide in the dopaminergic cell death process induced by 6-OHDA in primary cultures. Overall, this study demonstrates that the survival of the dopaminergic neurons can be highly increased by the adenoviral gene transfer of SOD1. An antioxidant gene transfer strategy using viral vectors expressing SOD1 is therefore potentially beneficial for protecting dopaminergic neurons in PD. [ABSTRACT FROM AUTHOR]
- Published
- 2002
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31. Adenovirus for neurodegenerative diseases: in vivo strategies and ex vivo gene therapy using human neural progenitors
- Author
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Sabaté O, Barkats M, Mh, Buc-Caron, Mn, Castel-Barthe, Finiels F, Philippe Horellou, Revah F, and Mallet J
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Neurons ,Neuroprotective Agents ,Genetic Vectors ,Nerve Degeneration ,Humans ,Genetic Therapy ,Nerve Growth Factors ,Adenoviridae ,Stem Cell Transplantation - Abstract
The discovery of major neurodegenerative mechanisms has opened the way to the development of novel therapeutic approaches. Gene therapy now enables researchers to overcome certain problems inherent to pharmacotherapy and to the grafting of embryonic cells. The production of recombinant adenoviruses are promising for in vivo gene therapy involving neuroprotective (Ad-SOD), neurotrophic (Ad-NGF) as well as restorative (Ad-TH) strategies. In addition, human neural progenitors offer great potential as vehicles for ex vivo gene therapy to replace degenerated cells in advanced stages of neurodegenerative diseases. This paper describes the clinical values of the new generations of adenoviral vectors.
32. Effect of Long-term Treatment with EGb 761 on Age-dependent Structural Changes in the Hippocampi of Three Inbred Mouse Strains
- Author
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Barkats, M., Venault, P., Christen, Y., and Cohen-Salmon, C.
- Published
- 1994
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33. Correction to: Intraneuronal aggregation of the β‑CTF fragment of APP (C99) induces Aβ‑independent lysosomal-autophagic pathology.
- Author
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Lauritzen I, Pardossi-Piquard R, Bourgeois A, Pagnotta S, Biferi MG, Barkats M, Lacor P, Klein W, Bauer C, and Checler F
- Published
- 2023
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34. Splicing efficiency of minor introns in a mouse model of SMA predominantly depends on their branchpoint sequence and can involve the contribution of major spliceosome components.
- Author
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Jacquier V, Prévot M, Gostan T, Bordonné R, Benkhelifa-Ziyyat S, Barkats M, and Soret J
- Subjects
- Animals, HeLa Cells, Humans, Introns, Mice, Muscular Atrophy, Spinal metabolism, RNA Splice Sites, Ribonucleoproteins, Small Nuclear metabolism, Muscular Atrophy, Spinal genetics, RNA Splicing, Spliceosomes metabolism
- Abstract
Spinal muscular atrophy (SMA) is a devastating neurodegenerative disease caused by reduced amounts of the ubiquitously expressed Survival of Motor Neuron (SMN) protein. In agreement with its crucial role in the biogenesis of spliceosomal snRNPs, SMN-deficiency is correlated to numerous splicing alterations in patient cells and various tissues of SMA mouse models. Among the snRNPs whose assembly is impacted by SMN-deficiency, those involved in the minor spliceosome are particularly affected. Importantly, splicing of several, but not all U12-dependent introns has been shown to be affected in different SMA models. Here, we have investigated the molecular determinants of this differential splicing in spinal cords from SMA mice. We show that the branchpoint sequence (BPS) is a key element controlling splicing efficiency of minor introns. Unexpectedly, splicing of several minor introns with suboptimal BPS is not affected in SMA mice. Using in vitro splicing experiments and oligonucleotides targeting minor or major snRNAs, we show for the first time that splicing of these introns involves both the minor and major machineries. Our results strongly suggest that splicing of a subset of minor introns is not affected in SMA mice because components of the major spliceosome compensate for the loss of minor splicing activity., (© 2022 Jacquier et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.)
- Published
- 2022
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35. Systemic Treatment of Fabry Disease Using a Novel AAV9 Vector Expressing α-Galactosidase A.
- Author
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Biferi MG, Cohen-Tannoudji M, García-Silva A, Souto-Rodríguez O, Viéitez-González I, San-Millán-Tejado B, Fernández-Carrera A, Pérez-Márquez T, Teijeira-Bautista S, Barrera S, Domínguez V, Marais T, González-Fernández Á, Barkats M, and Ortolano S
- Abstract
Fabry disease is a rare X-linked disorder affecting α-galactosidase A, a rate-limiting enzyme in lysosomal catabolism of glycosphingolipids. Current treatments present important limitations, such as low half-life and limited distribution, which gene therapy can overcome. The aim of this work was to test a novel adeno-associated viral vector, serotype 9 (AAV9), ubiquitously expressing human α-galactosidase A to treat Fabry disease (scAAV9-PGK-GLA). The vector was preliminary tested in newborns of a Fabry disease mouse model. 5 months after treatment, α-galactosidase A activity was detectable in the analyzed tissues, including the central nervous system. Moreover, we tested the vector in adult animals of both sexes at two doses and disease stages (presymptomatic and symptomatic) by single intravenous injection. We found that the exogenous α-galactosidase A was active in peripheral tissues as well as the central nervous system and prevented glycosphingolipid accumulation in treated animals up to 5 months following injection. Antibodies against α-galactosidase A were produced in 9 out of 32 treated animals, although enzyme activity in tissues was not significantly affected. These results demonstrate that scAAV9-PGK-GLA can drive widespread and sustained expression of α-galactosidase A, cross the blood brain barrier after systemic delivery, and reduce pathological signs of the Fabry disease mouse model., Competing Interests: The authors declare no competing interests., (© 2020 The Author(s).)
- Published
- 2020
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36. AAV9-Mediated Expression of SMN Restricted to Neurons Does Not Rescue the Spinal Muscular Atrophy Phenotype in Mice.
- Author
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Besse A, Astord S, Marais T, Roda M, Giroux B, Lejeune FX, Relaix F, Smeriglio P, Barkats M, and Biferi MG
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- Animals, Disease Models, Animal, Gene Expression, Gene Transfer Techniques, Genetic Therapy, Locomotion, Mice, Muscular Atrophy, Spinal drug therapy, Phenotype, Prognosis, Promoter Regions, Genetic, Spinal Cord metabolism, Spinal Cord pathology, Survival of Motor Neuron 1 Protein metabolism, Transduction, Genetic, Treatment Outcome, Dependovirus genetics, Genetic Vectors genetics, Motor Neurons metabolism, Motor Neurons virology, Muscular Atrophy, Spinal genetics, Survival of Motor Neuron 1 Protein genetics
- Abstract
Spinal muscular atrophy (SMA) is a neuromuscular disease mainly caused by mutations or deletions in the survival of motor neuron 1 (SMN1) gene and characterized by the degeneration of motor neurons and progressive muscle weakness. A viable therapeutic approach for SMA patients is a gene replacement strategy that restores functional SMN expression using adeno-associated virus serotype 9 (AAV9) vectors. Currently, systemic or intra-cerebrospinal fluid (CSF) delivery of AAV9-SMN is being explored in clinical trials. In this study, we show that the postnatal delivery of an AAV9 that expresses SMN under the control of the neuron-specific promoter synapsin selectively targets neurons without inducing re-expression in the peripheral organs of SMA mice. However, this approach is less efficient in restoring the survival and neuromuscular functions of SMA mice than the systemic or intra-CSF delivery of an AAV9 in which SMN is placed under the control of a ubiquitous promoter. This study suggests that further efforts are needed to understand the extent to which SMN is required in neurons and peripheral organs for a successful therapeutic effect., (Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2020
- Full Text
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37. [SMA: from gene discovery to gene therapy].
- Author
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Barkats M
- Subjects
- Animals, Child, Disease Models, Animal, Humans, Mice, Motor Neurons physiology, Muscular Atrophy, Spinal diagnosis, Mutation, Spinal Muscular Atrophies of Childhood diagnosis, Spinal Muscular Atrophies of Childhood genetics, Spinal Muscular Atrophies of Childhood therapy, Genetic Association Studies methods, Genetic Association Studies trends, Genetic Therapy methods, Genetic Therapy trends, Muscular Atrophy, Spinal genetics, Muscular Atrophy, Spinal therapy
- Abstract
Spinal muscular atrophy (SMA) is the most common genetic disease leading to infant mortality. This neuro-muscular disorder is caused by the loss or mutation of the telomeric copy of the 'survival of motor neuron' (Smn) gene, termed SMN1. Loss of SMN1 leads to reduced SMN protein levels, inducing degeneration of motor neurons (MN) and progressive muscle weakness and atrophy. Gene therapy, consisting of reintroducing SMN1 in the MNs, is an attractive approach for SMA. We showed the most efficient rescue of SMA mice to date after a single intravenous injection of an AAV9 expressing SMN1, highlighting the considerable potential of this method for the treatment of human SMA. Recently, a startup led by the Dr Kaspar decided to test this experimental approach in children with SMA type 1. Dr Mendell, in charge of this clinical project, showed a very significant increase of the lifespan and motor function of the patients (until 4 years) after a single injection of AAV9-SMN1 (named ZolgenSMA®) into an arm or leg vein. This gene therapy treatment obtained a marketing authorization by the FDA in May 24 and is now the first efficient therapy for neuromuscular disease., (© 2020 médecine/sciences – Inserm.)
- Published
- 2020
- Full Text
- View/download PDF
38. Targeting γ-secretase triggers the selective enrichment of oligomeric APP-CTFs in brain extracellular vesicles from Alzheimer cell and mouse models.
- Author
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Lauritzen I, Bécot A, Bourgeois A, Pardossi-Piquard R, Biferi MG, Barkats M, and Checler F
- Abstract
Background: We recently demonstrated an endolysosomal accumulation of the β-secretase-derived APP C-terminal fragment (CTF) C99 in brains of Alzheimer disease (AD) mouse models. Moreover, we showed that the treatment with the γ-secretase inhibitor (D6) led to further increased endolysosomal APP-CTF levels, but also revealed extracellular APP-CTF-associated immunostaining. We here hypothesized that this latter staining could reflect extracellular vesicle (EV)-associated APP-CTFs and aimed to characterize these γ-secretase inhibitor-induced APP-CTFs., Methods: EVs were purified from cell media or mouse brains from vehicle- or D6-treated C99 or APP
swedish expressing cells/mice and analyzed for APP-CTFs by immunoblot. Combined pharmacological, immunological and genetic approaches (presenilin invalidation and C99 dimerization mutants (GXXXG)) were used to characterize vesicle-containing APP-CTFs. Subcellular APP-CTF localization was determined by immunocytochemistry., Results: Purified EVs from both AD cell or mouse models were enriched in APP-CTFs as compared to EVs from control cells/brains. Surprisingly, EVs from D6-treated cells not only displayed increased C99 and C99-derived C83 levels but also higher molecular weight (HMW) APP-CTF-immunoreactivities that were hardly detectable in whole cell extracts. Accordingly, the intracellular levels of HMW APP-CTFs were amplified by the exosomal inhibitor GW4869. By combined pharmacological, immunological and genetic approaches, we established that these HMW APP-CTFs correspond to oligomeric APP-CTFs composed of C99 and/or C83. Immunocytochemical analysis showed that monomers were localized mainly to the trans -Golgi network, whereas oligomers were confined to endosomes and lysosomes, thus providing an anatomical support for the selective recovery of HMW APP-CTFs in EVs. The D6-induced APP-CTF oligomerization and subcellular mislocalization was indeed due to γ-secretase blockade, since it similarly occurred in presenilin-deficient fibroblasts. Further, our data proposed that besides favoring APP-CTF oligomerization by preventing C99 proteolysis, γ-secretase inhibiton also led to a defective SorLA-mediated retrograde transport of HMW APP-CTFs from endosomal compartments to the TGN., Conclusions: This is the first study to demonstrate the presence of oligomeric APP-CTFs in AD mouse models, the levels of which are selectively enriched in endolysosomal compartments including exosomes and amplified by γ-secretase inhibition. Future studies should evaluate the putative contribution of these exosome-associated APP-CTFs in AD onset, progression and spreading., Competing Interests: Competing interestsThe authors declare that they have no competing interests., (© The Author(s). 2019.)- Published
- 2019
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39. Intravenous administration of scAAV9-Hexb normalizes lifespan and prevents pathology in Sandhoff disease mice.
- Author
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Niemir N, Rouvière L, Besse A, Vanier MT, Dmytrus J, Marais T, Astord S, Puech JP, Panasyuk G, Cooper JD, Barkats M, and Caillaud C
- Subjects
- Administration, Intravenous, Animals, Animals, Newborn, Brain metabolism, Disease Models, Animal, Female, G(M2) Ganglioside metabolism, Gangliosides metabolism, Hexosaminidase B genetics, Hexosaminidase B metabolism, Male, Mice, Mice, Inbred C57BL, Sandhoff Disease metabolism, Hexosaminidase B pharmacology, Sandhoff Disease drug therapy, Sandhoff Disease pathology
- Abstract
Sandhoff disease (SD) is a rare inherited disorder caused by a deficiency of β-hexosaminidase activity which is fatal because no effective treatment is available. A mouse model of Hexb deficiency reproduces the key pathognomonic features of SD patients with severe ubiquitous lysosomal dysfunction, GM2 accumulation, neuroinflammation and neurodegeneration, culminating in death at 4 months. Here, we show that a single intravenous neonatal administration of a self-complementary adeno-associated virus 9 vector (scAAV9) expressing the Hexb cDNA in SD mice is safe and sufficient to prevent disease development. Importantly, we demonstrate for the first time that this treatment results in a normal lifespan (over 700 days) and normalizes motor function assessed by a battery of behavioral tests, with scAAV9-treated SD mice being indistinguishable from wild-type littermates. Biochemical analyses in multiple tissues showed a significant increase in hexosaminidase A activity, which reached 10-15% of normal levels. AAV9 treatment was sufficient to prevent GM2 and GA2 storage almost completely in the cerebrum (less so in the cerebellum), as well as thalamic reactive gliosis and thalamocortical neuron loss in treated Hexb-/- mice. In summary, this study demonstrated a widespread protective effect throughout the entire CNS after a single intravenous administration of the scAAV9-Hexb vector to neonatal SD mice.
- Published
- 2018
- Full Text
- View/download PDF
40. β-Amyloid Precursor Protein Intracellular Domain Controls Mitochondrial Function by Modulating Phosphatase and Tensin Homolog-Induced Kinase 1 Transcription in Cells and in Alzheimer Mice Models.
- Author
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Goiran T, Duplan E, Chami M, Bourgeois A, El Manaa W, Rouland L, Dunys J, Lauritzen I, You H, Stambolic V, Biféri MG, Barkats M, Pimplikar SW, Sergeant N, Colin M, Morais VA, Pardossi-Piquard R, Checler F, and Alves da Costa C
- Subjects
- Animals, Cell Line, Disease Models, Animal, Embryo, Mammalian, Fibroblasts, HEK293 Cells, Humans, Intracellular Space metabolism, Male, Mice, Mice, 129 Strain, Mice, Inbred C57BL, Mice, Transgenic, Alzheimer Disease metabolism, Amyloid Precursor Protein Secretases metabolism, Amyloid beta-Protein Precursor metabolism, Forkhead Box Protein O3 metabolism, Hippocampus metabolism, Mitochondria metabolism, Presenilins metabolism, Protein Kinases metabolism, Ubiquitin-Protein Ligases metabolism
- Abstract
Background: Mitophagy and mitochondrial dynamics alterations are two major hallmarks of neurodegenerative diseases. Dysfunctional mitochondria accumulate in Alzheimer's disease-affected brains by yet unexplained mechanisms., Methods: We combined cell biology, molecular biology, and pharmacological approaches to unravel a novel molecular pathway by which presenilins control phosphatase and tensin homolog-induced kinase 1 (Pink-1) expression and transcription. In vivo approaches were carried out on various transgenic and knockout animals as well as in adeno-associated virus-infected mice. Functional readout and mitochondrial physiology (mitochondrial potential) were assessed by combined procedures including flow cytometry, live imaging analysis, and immunohistochemistry., Results: We show that presenilins 1 and 2 trigger opposite effects on promoter transactivation, messenger RNA, and protein expression of Pink-1. This control is linked to γ-secretase activity and β-amyloid precursor protein but is independent of phosphatase and tensin homolog. We show that amyloid precursor protein intracellular domain (AICD) accounts for presenilin-dependent phenotype and upregulates Pink-1 transactivation in cells as well as in vivo in a Forkhead box O3a-dependent manner. Interestingly, the modulation of γ-secretase activity or AICD expression affects Pink-1-related control of mitophagy and mitochondrial dynamics. Finally, we show that parkin acts upstream of presenilins to control Pink-1 promoter transactivation and protein expression., Conclusions: Overall, we delineate a molecular cascade presenilins-AICD-Forkhead box O3a linking parkin to Pink-1. Our study demonstrates AICD-mediated Pink-1-dependent control of mitochondrial physiology by presenilins. Furthermore, it unravels a parkin-Pink-1 feedback loop controlling mitochondrial physiology that could be disrupted in neurodegenerative conditions., (Copyright © 2017 Society of Biological Psychiatry. Published by Elsevier Inc. All rights reserved.)
- Published
- 2018
- Full Text
- View/download PDF
41. Rescue of Pompe disease in mice by AAV-mediated liver delivery of secretable acid α-glucosidase.
- Author
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Puzzo F, Colella P, Biferi MG, Bali D, Paulk NK, Vidal P, Collaud F, Simon-Sola M, Charles S, Hardet R, Leborgne C, Meliani A, Cohen-Tannoudji M, Astord S, Gjata B, Sellier P, van Wittenberghe L, Vignaud A, Boisgerault F, Barkats M, Laforet P, Kay MA, Koeberl DD, Ronzitti G, and Mingozzi F
- Subjects
- Animals, Genetic Therapy, Genetic Vectors, Male, Mice, Mice, Knockout, Muscle, Skeletal metabolism, alpha-Glucosidases genetics, alpha-Glucosidases physiology, Dependovirus genetics, Glycogen Storage Disease Type II therapy, Liver metabolism
- Abstract
Glycogen storage disease type II or Pompe disease is a severe neuromuscular disorder caused by mutations in the lysosomal enzyme, acid α-glucosidase (GAA), which result in pathological accumulation of glycogen throughout the body. Enzyme replacement therapy is available for Pompe disease; however, it has limited efficacy, has high immunogenicity, and fails to correct pathological glycogen accumulation in nervous tissue and skeletal muscle. Using bioinformatics analysis and protein engineering, we developed transgenes encoding GAA that could be expressed and secreted by hepatocytes. Then, we used adeno-associated virus (AAV) vectors optimized for hepatic expression to deliver the GAA transgenes to Gaa knockout (Gaa
-/- ) mice, a model of Pompe disease. Therapeutic gene transfer to the liver rescued glycogen accumulation in muscle and the central nervous system, and ameliorated cardiac hypertrophy as well as muscle and respiratory dysfunction in the Gaa-/- mice; mouse survival was also increased. Secretable GAA showed improved therapeutic efficacy and lower immunogenicity compared to nonengineered GAA. Scale-up to nonhuman primates, and modeling of GAA expression in primary human hepatocytes using hepatotropic AAV vectors, demonstrated the therapeutic potential of AAV vector-mediated liver expression of secretable GAA for treating pathological glycogen accumulation in multiple tissues in Pompe disease., (Copyright © 2017 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)- Published
- 2017
- Full Text
- View/download PDF
42. A New AAV10-U7-Mediated Gene Therapy Prolongs Survival and Restores Function in an ALS Mouse Model.
- Author
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Biferi MG, Cohen-Tannoudji M, Cappelletto A, Giroux B, Roda M, Astord S, Marais T, Bos C, Voit T, Ferry A, and Barkats M
- Subjects
- Age of Onset, Amyotrophic Lateral Sclerosis mortality, Amyotrophic Lateral Sclerosis physiopathology, Amyotrophic Lateral Sclerosis therapy, Animals, Disease Models, Animal, Exons, Gene Order, Gene Transfer Techniques, Genetic Vectors administration & dosage, Humans, Mice, Mice, Transgenic, Motor Activity genetics, Oligonucleotides, Antisense, RNA Splice Sites, RNA, Messenger genetics, RNA, Messenger metabolism, Recovery of Function, Superoxide Dismutase-1 metabolism, Survival Rate, Transduction, Genetic, Amyotrophic Lateral Sclerosis genetics, Dependovirus genetics, Genetic Therapy methods, Genetic Vectors genetics, Superoxide Dismutase-1 genetics
- Abstract
One of the most promising therapeutic approaches for familial amyotrophic lateral sclerosis linked to superoxide dismutase 1 (SOD1) is the suppression of toxic mutant SOD1 in the affected tissues. Here, we report an innovative molecular strategy for inducing substantial, widespread, and sustained reduction of mutant human SOD1 (hSOD1) levels throughout the body of SOD1
G93A mice, leading to therapeutic effects in animals. Adeno-associated virus serotype rh10 vectors (AAV10) were used to mediate exon skipping of the hSOD1 pre-mRNA by expression of exon-2-targeted antisense sequences embedded in a modified U7 small-nuclear RNA (AAV10-U7-hSOD). Skipping of hSOD1 exon 2 led to the generation of a premature termination codon, inducing production of a deleted transcript that was subsequently degraded by the activation of nonsense-mediated decay. Combined intravenous and intracerebroventricular delivery of AAV10-U7-hSOD increased the survival of SOD1G93A mice injected either at birth or at 50 days of age (by 92% and 58%, respectively) and prevented weight loss and the decline of neuromuscular function. This study reports the effectiveness of an exon-skipping approach in SOD1-ALS mice, supporting the translation of this technology to the treatment of this as yet incurable disease., (Copyright © 2017 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.)- Published
- 2017
- Full Text
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43. SECIS-binding protein 2 interacts with the SMN complex and the methylosome for selenoprotein mRNP assembly and translation.
- Author
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Gribling-Burrer AS, Leichter M, Wurth L, Huttin A, Schlotter F, Troffer-Charlier N, Cura V, Barkats M, Cavarelli J, Massenet S, and Allmang C
- Subjects
- Glutathione Peroxidase, HEK293 Cells, HeLa Cells, Humans, Methylation, Models, Biological, Muscular Atrophy, Spinal metabolism, Muscular Atrophy, Spinal pathology, Protein Binding, Spinal Cord metabolism, Glutathione Peroxidase GPX1, Protein Biosynthesis, RNA-Binding Proteins metabolism, Ribonucleoproteins metabolism, SMN Complex Proteins metabolism, Selenoproteins metabolism
- Abstract
Selenoprotein synthesis requires the co-translational recoding of a UGASec codon. This process involves an RNA structural element, called Selenocysteine Insertion Sequence (SECIS) and the SECIS binding protein 2 (SBP2). Several selenoprotein mRNAs undergo unusual cap hypermethylation by the trimethylguanosine synthase 1 (Tgs1), which is recruited by the ubiquitous Survival of MotoNeurons (SMN) protein. SMN, the protein involved in spinal muscular atrophy, is part of a chaperone complex that collaborates with the methylosome for RNP assembly. Here, we analyze the role of individual SMN and methylosome components in selenoprotein mRNP assembly and translation. We show that SBP2 interacts directly with four proteins of the SMN complex and the methylosome core proteins. Nevertheless, SBP2 is not a methylation substrate of the methylosome. We found that both SMN and methylosome complexes are required for efficient translation of the selenoprotein GPx1 in vivo. We establish that the steady-state level of several selenoprotein mRNAs, major regulators of oxidative stress damage in neurons, is specifically reduced in the spinal cord of SMN-deficient mice and that cap hypermethylation of GPx1 mRNA is affected. Altogether we identified a new function of the SMN complex and the methylosome in selenoprotein mRNP assembly and expression., (© The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.)
- Published
- 2017
- Full Text
- View/download PDF
44. A codon-optimized Mecp2 transgene corrects breathing deficits and improves survival in a mouse model of Rett syndrome.
- Author
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Matagne V, Ehinger Y, Saidi L, Borges-Correia A, Barkats M, Bartoli M, Villard L, and Roux JC
- Subjects
- Amines, Animals, Apnea metabolism, Apnea pathology, Apnea prevention & control, Codon, Cyclohexanecarboxylic Acids, Dependovirus, Disease Models, Animal, Disease Progression, Gabapentin, Genetic Vectors, Green Fluorescent Proteins genetics, Green Fluorescent Proteins metabolism, Male, Mesencephalon metabolism, Mesencephalon pathology, Methyl-CpG-Binding Protein 2 genetics, Methyl-CpG-Binding Protein 2 metabolism, Mice, Inbred C57BL, Mice, Knockout, Respiration, Rett Syndrome metabolism, Rett Syndrome pathology, Survival Analysis, Tyrosine 3-Monooxygenase metabolism, Weight Gain, gamma-Aminobutyric Acid, Genetic Therapy, Methyl-CpG-Binding Protein 2 administration & dosage, Rett Syndrome therapy
- Abstract
Rett syndrome (RTT) is a severe X-linked neurodevelopmental disorder that is primarily caused by mutations in the methyl CpG binding protein 2 gene (MECP2). RTT is the second most prevalent cause of intellectual disability in girls and there is currently no cure for the disease. The finding that the deficits caused by the loss of Mecp2 are reversible in the mouse has bolstered interest in gene therapy as a cure for RTT. In order to assess the feasibility of gene therapy in a RTT mouse model, and in keeping with translational goals, we investigated the efficacy of a self-complementary AAV9 vector expressing a codon-optimized version of Mecp2 (AAV9-MCO) delivered via a systemic approach in early symptomatic Mecp2-deficient (KO) mice. Our results show that AAV9-MCO administered at a dose of 2×10
11 viral genome (vg)/mouse was able to significantly increase survival and weight gain, and delay the occurrence of behavioral deficits. Apneas, which are one of the core RTT breathing deficits, were significantly decreased to WT levels in Mecp2 KO mice after AAV9-MCO administration. Semi-quantitative analysis showed that AAV9-MCO administration in Mecp2 KO mice resulted in 10 to 20% Mecp2 immunopositive cells compared to WT animals, with the highest Mecp2 expression found in midbrain regions known to regulate cardio-respiratory functions. In addition, we also found a cell autonomous increase in tyrosine hydroxylase levels in the A1C1 and A2C2 catecholaminergic Mecp2+ neurons in treated Mecp2 KO mice, which may partly explain the beneficial effect of AAV9-MCO administration on apneas occurrence., (Copyright © 2016 Elsevier Inc. All rights reserved.)- Published
- 2017
- Full Text
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45. Efficacy and biodistribution analysis of intracerebroventricular administration of an optimized scAAV9-SMN1 vector in a mouse model of spinal muscular atrophy.
- Author
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Armbruster N, Lattanzi A, Jeavons M, Van Wittenberghe L, Gjata B, Marais T, Martin S, Vignaud A, Voit T, Mavilio F, Barkats M, and Buj-Bello A
- Abstract
Spinal muscular atrophy (SMA) is an autosomal recessive disease of variable severity caused by mutations in the SMN1 gene. Deficiency of the ubiquitous SMN function results in spinal cord α-motor neuron degeneration and proximal muscle weakness. Gene replacement therapy with recombinant adeno-associated viral (AAV) vectors showed therapeutic efficacy in several animal models of SMA. Here, we report a study aimed at analyzing the efficacy and biodistribution of a serotype-9, self-complementary AAV vector expressing a codon-optimized human SMN1 coding sequence (coSMN1) under the control of the constitutive phosphoglycerate kinase (PGK) promoter in neonatal SMNΔ7 mice, a severe animal model of the disease. We administered the scAAV9-coSMN1 vector in the intracerebroventricular (ICV) space in a dose-escalating mode, and analyzed survival, vector biodistribution and SMN protein expression in the spinal cord and peripheral tissues. All treated mice showed a significant, dose-dependent rescue of lifespan and growth with a median survival of 346 days. Additional administration of vector by an intravenous route (ICV+IV) did not improve survival, and vector biodistribution analysis 90 days postinjection indicated that diffusion from the cerebrospinal fluid to the periphery was sufficient to rescue the SMA phenotype. These results support the preclinical development of SMN1 gene therapy by CSF vector delivery.
- Published
- 2016
- Full Text
- View/download PDF
46. Intraneuronal aggregation of the β-CTF fragment of APP (C99) induces Aβ-independent lysosomal-autophagic pathology.
- Author
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Lauritzen I, Pardossi-Piquard R, Bourgeois A, Pagnotta S, Biferi MG, Barkats M, Lacor P, Klein W, Bauer C, and Checler F
- Subjects
- Alzheimer Disease metabolism, Amyloid beta-Peptides metabolism, Animals, Autophagy physiology, Brain metabolism, Disease Models, Animal, Endosomes metabolism, Lysosomes metabolism, Mice, Inbred C57BL, Mice, Transgenic, Neurons pathology, Alzheimer Disease pathology, Amyloid Precursor Protein Secretases metabolism, Amyloid beta-Protein Precursor metabolism, Brain pathology, Neurons metabolism
- Abstract
Endosomal-autophagic-lysosomal (EAL) dysfunction is an early and prominent neuropathological feature of Alzheimers's disease, yet the exact molecular mechanisms contributing to this pathology remain undefined. By combined biochemical, immunohistochemical and ultrastructural approaches, we demonstrate a link between EAL pathology and the intraneuronal accumulation of the β-secretase-derived βAPP fragment (C99) in two in vivo models, 3xTgAD mice and adeno-associated viral-mediated C99-infected mice. We present a pathological loop in which the accumulation of C99 is both the effect and causality of impaired lysosomal-autophagic function. The deleterious effect of C99 was found to be linked to its aggregation within EAL-vesicle membranes leading to disrupted lysosomal proteolysis and autophagic impairment. This effect was Aβ independent and was even exacerbated when γ-secretase was pharmacologically inhibited. No effect was observed in inhibitor-treated wild-type animals suggesting that lysosomal dysfunction was indeed directly linked to C99 accumulation. In some brain areas, strong C99 expression also led to inflammatory responses and synaptic dysfunction. Taken together, this work demonstrates a toxic effect of C99 which could underlie some of the early-stage anatomical hallmarks of Alzheimer's disease pathology. Our work also proposes molecular mechanisms likely explaining some of the unfavorable side-effects associated with γ-secretase inhibitor-directed therapies.
- Published
- 2016
- Full Text
- View/download PDF
47. Lentiviral vector-mediated overexpression of mutant ataxin-7 recapitulates SCA7 pathology and promotes accumulation of the FUS/TLS and MBNL1 RNA-binding proteins.
- Author
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Alves S, Marais T, Biferi MG, Furling D, Marinello M, El Hachimi K, Cartier N, Ruberg M, Stevanin G, Brice A, Barkats M, and Sittler A
- Subjects
- Animals, Ataxin-7 genetics, DNA-Binding Proteins genetics, Disease Models, Animal, Female, Humans, Lentivirus genetics, Mice, Inbred C57BL, Neurons metabolism, Phenotype, Ataxin-7 metabolism, DNA-Binding Proteins metabolism, RNA-Binding Proteins metabolism, Spinocerebellar Ataxias genetics
- Abstract
Background: We used lentiviral vectors (LVs) to generate a new SCA7 animal model overexpressing a truncated mutant ataxin-7 (MUT ATXN7) fragment in the mouse cerebellum, in order to characterize the specific neuropathological and behavioral consequences of the genetic defect in this brain structure., Results: LV-mediated overexpression of MUT ATXN7 into the cerebellum of C57/BL6 adult mice induced neuropathological features similar to that observed in patients, such as intranuclear aggregates in Purkinje cells (PC), loss of synaptic markers, neuroinflammation, and neuronal death. No neuropathological changes were observed when truncated wild-type ataxin-7 (WT ATXN7) was injected. Interestingly, the local delivery of LV-expressing mutant ataxin-7 (LV-MUT-ATXN7) into the cerebellum of wild-type mice also mediated the development of an ataxic phenotype at 8 to 12 weeks post-injection. Importantly, our data revealed abnormal levels of the FUS/TLS, MBNL1, and TDP-43 RNA-binding proteins in the cerebellum of the LV-MUT-ATXN7 injected mice. MUT ATXN7 overexpression induced an increase in the levels of the pathological phosphorylated TDP-43, and a decrease in the levels of soluble FUS/TLS, with both proteins accumulating within ATXN7-positive intranuclear inclusions. MBNL1 also co-aggregated with MUT ATXN7 in most PC nuclear inclusions. Interestingly, no MBNL2 aggregation was observed in cerebellar MUT ATXN7 aggregates. Immunohistochemical studies in postmortem tissue from SCA7 patients and SCA7 knock-in mice confirmed SCA7-induced nuclear accumulation of FUS/TLS and MBNL1, strongly suggesting that these proteins play a physiopathological role in SCA7., Conclusions: This study validates a novel SCA7 mouse model based on lentiviral vectors, in which strong and sustained expression of MUT ATXN7 in the cerebellum was found sufficient to generate motor defects.
- Published
- 2016
- Full Text
- View/download PDF
48. Estrogen-mediated downregulation of AIRE influences sexual dimorphism in autoimmune diseases.
- Author
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Dragin N, Bismuth J, Cizeron-Clairac G, Biferi MG, Berthault C, Serraf A, Nottin R, Klatzmann D, Cumano A, Barkats M, Le Panse R, and Berrih-Aknin S
- Subjects
- Adolescent, Adult, Animals, Autoimmune Diseases genetics, Cells, Cultured, Child, Child, Preschool, CpG Islands, DNA Methylation, Estrogen Receptor alpha genetics, Estrogen Receptor alpha metabolism, Estrogens genetics, Female, Humans, Infant, Male, Mice, Mice, Inbred C3H, Middle Aged, Thymus Gland metabolism, Transcription Factors genetics, AIRE Protein, Autoimmune Diseases metabolism, Estrogens metabolism, Gene Expression Regulation, Sex Characteristics, Transcription Factors biosynthesis
- Abstract
Autoimmune diseases affect 5% to 8% of the population, and females are more susceptible to these diseases than males. Here, we analyzed human thymic transcriptome and revealed sex-associated differences in the expression of tissue-specific antigens that are controlled by the autoimmune regulator (AIRE), a key factor in central tolerance. We hypothesized that the level of AIRE is linked to sexual dimorphism susceptibility to autoimmune diseases. In human and mouse thymus, females expressed less AIRE (mRNA and protein) than males after puberty. These results were confirmed in purified murine thymic epithelial cells (TECs). We also demonstrated that AIRE expression is related to sexual hormones, as male castration decreased AIRE thymic expression and estrogen receptor α-deficient mice did not show a sex disparity for AIRE expression. Moreover, estrogen treatment resulted in downregulation of AIRE expression in cultured human TECs, human thymic tissue grafted to immunodeficient mice, and murine fetal thymus organ cultures. AIRE levels in human thymus grafted in immunodeficient mice depended upon the sex of the recipient. Estrogen also upregulated the number of methylated CpG sites in the AIRE promoter. Together, our results indicate that in females, estrogen induces epigenetic changes in the AIRE gene, leading to reduced AIRE expression under a threshold that increases female susceptibility to autoimmune diseases.
- Published
- 2016
- Full Text
- View/download PDF
49. Moving towards treatments for spinal muscular atrophy: hopes and limits.
- Author
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Wirth B, Barkats M, Martinat C, Sendtner M, and Gillingwater TH
- Subjects
- Animals, Dependovirus genetics, Disease Models, Animal, Genetic Vectors, Humans, Infant, Newborn, Muscular Atrophy, Spinal genetics, Muscular Atrophy, Spinal physiopathology, Survival of Motor Neuron 1 Protein genetics, Survival of Motor Neuron 2 Protein genetics, Genetic Therapy methods, Muscular Atrophy, Spinal therapy
- Abstract
Spinal muscular atrophy (SMA), one of the most frequent and devastating genetic disorders causing neuromuscular degeneration, has reached the forefront of clinical translation. The quite unique genetic situation of SMA patients, who lack functional SMN1 but carry the misspliced SMN2 copy gene, creates the possibility of correcting SMN2 splicing by antisense oligonucleotides or drugs. Both strategies showed impressive results in pre-clinical trials and are now in Phase II-III clinical trials. SMN gene therapy approaches using AAV9-SMN vectors are also highly promising and have entered a Phase I clinical trial. However, careful analysis of SMA animal models and patients has revealed some limitations that need to be taken very seriously, including: i) a limited time-window for successful therapy delivery, making neonatal screening of SMA mandatory; ii) multi-organ impairment, requiring systemic delivery of therapies; and iii) a potential need for combined therapies that both increase SMN levels and target pathways that preserve/rescue motor neuron function over the lifespan. Meeting these challenges will likely be crucial to cure SMA, instead of only ameliorating symptoms, particularly in its most severe form. This review discusses therapies currently in clinical trials, the hopes for SMA therapy, and the potential limitations of these new approaches.
- Published
- 2015
- Full Text
- View/download PDF
50. Systemic AAVrh10 provides higher transgene expression than AAV9 in the brain and the spinal cord of neonatal mice.
- Author
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Tanguy Y, Biferi MG, Besse A, Astord S, Cohen-Tannoudji M, Marais T, and Barkats M
- Abstract
Systemic delivery of self-complementary (sc) adeno-associated-virus vector of serotype 9 (AAV9) was recently shown to provide robust and widespread gene transfer to the central nervous system (CNS), opening new avenues for practical, and non-invasive gene therapy of neurological diseases. More recently, AAV of serotype rh10 (AAVrh10) was also found highly efficient to mediate CNS transduction after intravenous administration in mice. However, only a few studies compared AAV9 and AAVrh10 efficiencies, particularly in the spinal cord. In this study, we compared the transduction capabilities of AAV9 and AAVrh10 in the brain, the spinal cord, and the peripheral nervous system (PNS) after intravenous delivery in neonatal mice. As reported in previous studies, AAVrh10 achieved either similar or higher transduction than AAV9 in all the examined brain regions. The superiority of AAVrh10 over AAV9 appeared statistically significant only in the medulla and the cerebellum, but a clear trend was also observed in other structures like the hippocampus or the cortex. In contrast to previous studies, we found that AAVrh10 was more efficient than AAV9 for transduction of the dorsal spinal cord and the lower motor neurons (MNs). However, differences between the two serotypes appeared mainly significant at low dose, and surprisingly, increasing the dose did not improve AAVrh10 distribution in the spinal cord, in contrary to AAV9. Similar dose-related differences between transduction efficiency of the two serotypes were also observed in the sciatic nerve. These findings suggest differences in the transduction mechanisms of these two serotypes, which both hold great promise for gene therapy of neurological diseases.
- Published
- 2015
- Full Text
- View/download PDF
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