title: Dominant-Negative Effects of Adult-Onset Huntingtin Mutations Alter the Division of Human Embryonic Stem Cells-Derived Neural Cells.
abstract: Mutations of the huntingtin protein (HTT) gene underlie both adult-onset and juvenile forms of Huntington's disease (HD). HTT modulates mitotic spindle orientation and cell fate in mouse cortical progenitors from the ventricular zone. Using human embryonic stem cells (hESC) characterized as carrying mutations associated with adult-onset disease during pre-implantation genetic diagnosis, we investigated the influence of human HTT and of an adult-onset HD mutation on mitotic spindle orientation in human neural stem cells (NSCs) derived from hESCs. The RNAi-mediated silencing of both HTT alleles in neural stem cells derived from hESCs disrupted spindle orientation and led to the mislocalization of dynein, the p150Glued subunit of dynactin and the large nuclear mitotic apparatus (NuMA) protein. We also investigated the effect of the adult-onset HD mutation on the role of HTT during spindle orientation in NSCs derived from HD-hESCs. By combining SNP-targeting allele-specific silencing and gain-of-function approaches, we showed that a 46-glutamine expansion in human HTT was sufficient for a dominant-negative effect on spindle orientation and changes in the distribution within the spindle pole and the cell cortex of dynein, p150Glued and NuMA in neural cells. Thus, neural derivatives of disease-specific human pluripotent stem cells constitute a relevant biological resource for exploring the impact of adult-onset HD mutations of the HTT gene on the division of neural progenitors, with potential applications in HD drug discovery targeting HTT-dynein-p150Glued complex interactions.
authors: Grenoble Institut des Neurosciences, Grenoble, France.|INSERM U836, Grenoble, France.|Grenoble Alpes University, Grenoble, France.|CNC-Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal.|Institute for Interdisciplinary Research of the University of Coimbra (IIIUC), Coimbra, Portugal.
authors: Carla
authors: C
authors: Lopes
authors: CECS, I-STEM, AFM, Corbeil-Essonnes, France.
authors: Sophie
authors: S
authors: Aubert
authors: Inserm U861, I-STEM, AFM, Corbeil-Essonnes, France.|UEVE U861, I-STEM, AFM, Evry, France.
authors: Fany
authors: F
authors: Bourgois-Rocha
authors: Grenoble Institut des Neurosciences, Grenoble, France.|INSERM U836, Grenoble, France.|Grenoble Alpes University, Grenoble, France.
authors: Monia
authors: M
authors: Barnat
authors: CNC-Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal.|Faculty of Medicine, University of Coimbra, Coimbra, Portugal.
authors: Ana Cristina
authors: AC
authors: Rego
authors: Lausanne University Hospital (CHUV), Department of Clinical Neurosciences (DNC), Laboratory of Cellular and Molecular Neurotherapies (LNCM), Lausanne, Switzerland.
authors: Nicole
authors: N
authors: Déglon
authors: Inserm U861, I-STEM, AFM, Corbeil-Essonnes, France.|UEVE U861, I-STEM, AFM, Evry, France.
authors: Anselme L
authors: AL
authors: Perrier
authors: Grenoble Institut des Neurosciences, Grenoble, France.|INSERM U836, Grenoble, France.|Grenoble Alpes University, Grenoble, France.
authors: Sandrine
authors: S
authors: Humbert
mesh_terms: D000328:Adult; D017668:Age of Onset; D000483:Alleles; D034961:Antigens, Nuclear / Q000032:analysis; D018797:Cell Cycle Proteins; D002478:Cells, Cultured; D000072159:Dynactin Complex; D004398:Dyneins / Q000032:analysis; D005799:Genes, Dominant; D000066449:Human Embryonic Stem Cells / Q000166:cytology; D006801:Humans; D000071058:Huntingtin Protein; D008869:Microtubule-Associated Proteins / Q000032:analysis; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000502:physiology; D058953:Neural Stem Cells / Q000378:metabolism* / Q000648:ultrastructure; D034521:Nuclear Matrix-Associated Proteins / Q000032:analysis; D010455:Peptides / Q000032:analysis; D039904:Pluripotent Stem Cells / Q000166:cytology; D020641:Polymorphism, Single Nucleotide; D021381:Protein Transport; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D008941:Spindle Apparatus / Q000648:ultrastructure; D013347:Subcellular Fractions / Q000737:chemistry; D019680:Trinucleotide Repeat Expansion
chemical_list: D034961:Antigens, Nuclear; D018797:Cell Cycle Proteins; D000072159:Dynactin Complex; C086055:HTT protein, human; D000071058:Huntingtin Protein; D008869:Microtubule-Associated Proteins; C050045:NUMA1 protein, human; D009419:Nerve Tissue Proteins; D034521:Nuclear Matrix-Associated Proteins; D010455:Peptides; D034741:RNA, Small Interfering; C097188:polyglutamine; D004398:Dyneins