title: Allele-specific silencing of mutant huntingtin in rodent brain and human stem cells.
abstract: Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder resulting from polyglutamine expansion in the huntingtin (HTT) protein and for which there is no cure. Although suppression of both wild type and mutant HTT expression by RNA interference is a promising therapeutic strategy, a selective silencing of mutant HTT represents the safest approach preserving WT HTT expression and functions. We developed small hairpin RNAs (shRNAs) targeting single nucleotide polymorphisms (SNP) present in the HTT gene to selectively target the disease HTT isoform. Most of these shRNAs silenced, efficiently and selectively, mutant HTT in vitro. Lentiviral-mediated infection with the shRNAs led to selective degradation of mutant HTT mRNA and prevented the apparition of neuropathology in HD rat's striatum expressing mutant HTT containing the various SNPs. In transgenic BACHD mice, the mutant HTT allele was also silenced by this approach, further demonstrating the potential for allele-specific silencing. Finally, the allele-specific silencing of mutant HTT in human embryonic stem cells was accompanied by functional recovery of the vesicular transport of BDNF along microtubules. These findings provide evidence of the therapeutic potential of allele-specific RNA interference for HD.
authors.affiliation: Institute of Biomedical Imaging (I2BM) and Molecular Imaging Research Center (MIRCen), Atomic Energy Commission (CEA), Fontenay-aux-Roses, France; URA2210, Centre National de Recherché Scientifique (CNRS), Fontenay-aux-Roses, France.
authors.forename: Valérie
authors.initials: V
authors.lastname: Drouet
authors.affiliation: Institute of Biomedical Imaging (I2BM) and Molecular Imaging Research Center (MIRCen), Atomic Energy Commission (CEA), Fontenay-aux-Roses, France; URA2210, Centre National de Recherché Scientifique (CNRS), Fontenay-aux-Roses, France.
authors.forename: Marta
authors.initials: M
authors.lastname: Ruiz
authors.affiliation: Institut Curie, Orsay, France; UMR3306, Centre National de Recherché Scientifique (CNRS), Orsay, France; U1005, Institut National de la Santé et de la Recherche Médicale (INSERM), Orsay France.
authors.forename: Diana
authors.initials: D
authors.lastname: Zala
authors.affiliation: U861, Institut National de la Santé et de la Recherche Médicale (INSERM), AFM, Evry, France; UEVE U861, I-STEM, AFM, Evry, France.
authors.forename: Maxime
authors.initials: M
authors.lastname: Feyeux
authors.affiliation: Institute of Biomedical Imaging (I2BM) and Molecular Imaging Research Center (MIRCen), Atomic Energy Commission (CEA), Fontenay-aux-Roses, France; URA2210, Centre National de Recherché Scientifique (CNRS), Fontenay-aux-Roses, France.
authors.forename: Gwennaëlle
authors.initials: G
authors.lastname: Auregan
authors.affiliation: Institute of Biomedical Imaging (I2BM) and Molecular Imaging Research Center (MIRCen), Atomic Energy Commission (CEA), Fontenay-aux-Roses, France; URA2210, Centre National de Recherché Scientifique (CNRS), Fontenay-aux-Roses, France.
authors.forename: Karine
authors.initials: K
authors.lastname: Cambon
authors.affiliation: Department of Clinical Neurosciences (DNC), Lausanne University Hospital (CHUV), Lausanne, Switzerland.
authors.forename: Laetitia
authors.initials: L
authors.lastname: Troquier
authors.affiliation: Institute of Biomedical Imaging (I2BM) and Molecular Imaging Research Center (MIRCen), Atomic Energy Commission (CEA), Fontenay-aux-Roses, France; URA2210, Centre National de Recherché Scientifique (CNRS), Fontenay-aux-Roses, France.
authors.forename: Johann
authors.initials: J
authors.lastname: Carpentier
authors.affiliation: CECS, I-STEM, AFM, Evry, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Aubert
authors.affiliation: Department of Clinical Neurosciences (DNC), Lausanne University Hospital (CHUV), Lausanne, Switzerland.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Merienne
authors.affiliation: U861, Institut National de la Santé et de la Recherche Médicale (INSERM), AFM, Evry, France; UEVE U861, I-STEM, AFM, Evry, France.
authors.forename: Fany
authors.initials: F
authors.lastname: Bourgois-Rocha
authors.affiliation: Institute of Biomedical Imaging (I2BM) and Molecular Imaging Research Center (MIRCen), Atomic Energy Commission (CEA), Fontenay-aux-Roses, France; URA2210, Centre National de Recherché Scientifique (CNRS), Fontenay-aux-Roses, France.
authors.forename: Raymonde
authors.initials: R
authors.lastname: Hassig
authors.affiliation: Department of Clinical Neurosciences (DNC), Lausanne University Hospital (CHUV), Lausanne, Switzerland.
authors.forename: Maria
authors.initials: M
authors.lastname: Rey
authors.affiliation: Institute of Biomedical Imaging (I2BM) and Molecular Imaging Research Center (MIRCen), Atomic Energy Commission (CEA), Fontenay-aux-Roses, France; URA2210, Centre National de Recherché Scientifique (CNRS), Fontenay-aux-Roses, France.
authors.forename: Noëlle
authors.initials: N
authors.lastname: Dufour
authors.affiliation: Institut Curie, Orsay, France; UMR3306, Centre National de Recherché Scientifique (CNRS), Orsay, France; U1005, Institut National de la Santé et de la Recherche Médicale (INSERM), Orsay France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Saudou
authors.affiliation: U861, Institut National de la Santé et de la Recherche Médicale (INSERM), AFM, Evry, France; UEVE U861, I-STEM, AFM, Evry, France.
authors.forename: Anselme L
authors.initials: AL
authors.lastname: Perrier
authors.affiliation: Institute of Biomedical Imaging (I2BM) and Molecular Imaging Research Center (MIRCen), Atomic Energy Commission (CEA), Fontenay-aux-Roses, France; URA2210, Centre National de Recherché Scientifique (CNRS), Fontenay-aux-Roses, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Hantraye
authors.affiliation: Institute of Biomedical Imaging (I2BM) and Molecular Imaging Research Center (MIRCen), Atomic Energy Commission (CEA), Fontenay-aux-Roses, France; URA2210, Centre National de Recherché Scientifique (CNRS), Fontenay-aux-Roses, France; Department of Clinical Neurosciences (DNC), Lausanne University Hospital (CHUV), Lausanne, Switzerland.
authors.forename: Nicole
authors.initials: N
authors.lastname: Déglon
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology* / Q000378:metabolism; D002478:Cells, Cultured; D004195:Disease Models, Animal; D053595:Embryonic Stem Cells / Q000166:cytology; D015316:Genetic Therapy / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D000071058:Huntingtin Protein; D006816:Huntington Disease / Q000235:genetics / Q000628:therapy*; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D050505:Mutant Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide; D059368:RNA Isoforms / Q000378:metabolism; D020871:RNA Stability; D034741:RNA, Small Interfering / Q000235:genetics*; D051381:Rats; D017208:Rats, Wistar
chemical_list: C086055:HTT protein, human; D000071058:Huntingtin Protein; D050505:Mutant Proteins; D009419:Nerve Tissue Proteins; D059368:RNA Isoforms; D034741:RNA, Small Interfering