title: The first reported generation of several induced pluripotent stem cell lines from homozygous and heterozygous Huntington's disease patients demonstrates mutation related enhanced lysosomal activity.
abstract: Neuronal disorders, like Huntington's disease (HD), are difficult to study, due to limited cell accessibility, late onset manifestations, and low availability of material. The establishment of an in vitro model that recapitulates features of the disease may help understanding the cellular and molecular events that trigger disease manifestations. Here, we describe the generation and characterization of a series of induced pluripotent stem (iPS) cells derived from patients with HD, including two rare homozygous genotypes and one heterozygous genotype. We used lentiviral technology to transfer key genes for inducing reprogramming. To confirm pluripotency and differentiation of iPS cells, we used PCR amplification and immunocytochemistry to measure the expression of marker genes in embryoid bodies and neurons. We also analyzed teratomas that formed in iPS cell-injected mice. We found that the length of the pathological CAG repeat did not increase during reprogramming, after long term growth in vitro, and after differentiation into neurons. In addition, we observed no differences between normal and mutant genotypes in reprogramming, growth rate, caspase activation or neuronal differentiation. However, we observed a significant increase in lysosomal activity in HD-iPS cells compared to control iPS cells, both during self-renewal and in iPS-derived neurons. In conclusion, we have established stable HD-iPS cell lines that can be used for investigating disease mechanisms that underlie HD. The CAG stability and lysosomal activity represent novel observations in HD-iPS cells. In the future, these cells may provide the basis for a powerful platform for drug screening and target identification in HD.
authors.affiliation: Department of Pharmacological Sciences and Centre for Stem Cell Research, University of Milan, Milan, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Camnasio
authors.forename: Alessia
authors.initials: A
authors.lastname: Delli Carri
authors.forename: Angelo
authors.initials: A
authors.lastname: Lombardo
authors.forename: Iwona
authors.initials: I
authors.lastname: Grad
authors.forename: Caterina
authors.initials: C
authors.lastname: Mariotti
authors.forename: Alessia
authors.initials: A
authors.lastname: Castucci
authors.forename: Björn
authors.initials: B
authors.lastname: Rozell
authors.forename: Pietro
authors.initials: P
authors.lastname: Lo Riso
authors.forename: Valentina
authors.initials: V
authors.lastname: Castiglioni
authors.forename: Chiara
authors.initials: C
authors.lastname: Zuccato
authors.forename: Christelle
authors.initials: C
authors.lastname: Rochon
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Takashima
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Diaferia
authors.forename: Ida
authors.initials: I
authors.lastname: Biunno
authors.forename: Cinzia
authors.initials: C
authors.lastname: Gellera
authors.forename: Marisa
authors.initials: M
authors.lastname: Jaconi
authors.forename: Austin
authors.initials: A
authors.lastname: Smith
authors.forename: Outi
authors.initials: O
authors.lastname: Hovatta
authors.forename: Luigi
authors.initials: L
authors.lastname: Naldini
authors.forename: Stefano
authors.initials: S
authors.lastname: Di Donato
authors.forename: Anis
authors.initials: A
authors.lastname: Feki
authors.forename: Elena
authors.initials: E
authors.lastname: Cattaneo
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D000071058:Huntingtin Protein; D006816:Huntington Disease / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D008247:Lysosomes / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D016513:Mice, SCID; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000235:genetics* / Q000378:metabolism; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000378:metabolism*; D013724:Teratoma / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation / Q000502:physiology
chemical_list: C086055:HTT protein, human; D000071058:Huntingtin Protein; D009419:Nerve Tissue Proteins