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
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 m...
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| Format: | Article |
| Language: | English |
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Elsevier
2012-04-01
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| Series: | Neurobiology of Disease |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0969996111004177 |
| _version_ | 1831640080074145792 |
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| author | Stefano Camnasio Alessia Delli Carri Angelo Lombardo Iwona Grad Caterina Mariotti Alessia Castucci Björn Rozell Pietro Lo Riso Valentina Castiglioni Chiara Zuccato Christelle Rochon Yasuhiro Takashima Giuseppe Diaferia Ida Biunno Cinzia Gellera Marisa Jaconi Austin Smith Outi Hovatta Luigi Naldini Stefano Di Donato Anis Feki Elena Cattaneo |
| author_facet | Stefano Camnasio Alessia Delli Carri Angelo Lombardo Iwona Grad Caterina Mariotti Alessia Castucci Björn Rozell Pietro Lo Riso Valentina Castiglioni Chiara Zuccato Christelle Rochon Yasuhiro Takashima Giuseppe Diaferia Ida Biunno Cinzia Gellera Marisa Jaconi Austin Smith Outi Hovatta Luigi Naldini Stefano Di Donato Anis Feki Elena Cattaneo |
| author_sort | Stefano Camnasio |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-19T12:00:24Z |
| format | Article |
| id | doaj.art-23c2258d4ef340178a5b5ebcc449756a |
| institution | Directory Open Access Journal |
| issn | 1095-953X |
| language | English |
| last_indexed | 2024-12-19T12:00:24Z |
| publishDate | 2012-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Neurobiology of Disease |
| spelling | doaj.art-23c2258d4ef340178a5b5ebcc449756a2022-12-21T20:22:31ZengElsevierNeurobiology of Disease1095-953X2012-04-014614151The first reported generation of several induced pluripotent stem cell lines from homozygous and heterozygous Huntington's disease patients demonstrates mutation related enhanced lysosomal activityStefano Camnasio0Alessia Delli Carri1Angelo Lombardo2Iwona Grad3Caterina Mariotti4Alessia Castucci5Björn Rozell6Pietro Lo Riso7Valentina Castiglioni8Chiara Zuccato9Christelle Rochon10Yasuhiro Takashima11Giuseppe Diaferia12Ida Biunno13Cinzia Gellera14Marisa Jaconi15Austin Smith16Outi Hovatta17Luigi Naldini18Stefano Di Donato19Anis Feki20Elena Cattaneo21Department of Pharmacological Sciences and Centre for Stem Cell Research, University of Milan, Milan, ItalyDepartment of Pharmacological Sciences and Centre for Stem Cell Research, University of Milan, Milan, ItalySan Raffaele Telethon Institute of Gene Therapy and Vita Salute San Raffaele University, San Raffaele Scientific Institute, Milan, ItalyStem Cells Research Laboratory, Department of Obstetrics & Gynecology, Geneva University, Hospital Geneva, SwitzerlandUnit of Genetics of neurodegenerative and metabolic diseases, Carlo Besta Neurological Institute, Milan, ItalyUnit of Genetics of neurodegenerative and metabolic diseases, Carlo Besta Neurological Institute, Milan, ItalyDepartment of Veterinary Disease Biology, Faculty of Life Sciences, University of Copenhagen, Ridbanevej 3 1870 Frederiksberg C, DenmarkSan Raffaele Telethon Institute of Gene Therapy and Vita Salute San Raffaele University, San Raffaele Scientific Institute, Milan, ItalyDepartment of Pharmacological Sciences and Centre for Stem Cell Research, University of Milan, Milan, ItalyDepartment of Pharmacological Sciences and Centre for Stem Cell Research, University of Milan, Milan, ItalyDepartment of Pharmacological Sciences and Centre for Stem Cell Research, University of Milan, Milan, ItalyWellcome Trust Centre for Stem Cell Research and Department of Biochemistry, University of Cambridge, Cambridge, CB2 1QR, UKIntegrated Systems Engineering, s.r.l., via Fantoli 16/15, 20138 Milan, ItalyIntegrated Systems Engineering, s.r.l., via Fantoli 16/15, 20138 Milan, ItalyUnit of Genetics of neurodegenerative and metabolic diseases, Carlo Besta Neurological Institute, Milan, ItalyEmbryonic Stem Cell Laboratory Department of Pathology and Immunology, Faculty of Medicine, Geneva University, SwitzerlandWellcome Trust Centre for Stem Cell Research and Department of Biochemistry, University of Cambridge, Cambridge, CB2 1QR, UKDepartment of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, SwedenSan Raffaele Telethon Institute of Gene Therapy and Vita Salute San Raffaele University, San Raffaele Scientific Institute, Milan, ItalyUnit of Genetics of neurodegenerative and metabolic diseases, Carlo Besta Neurological Institute, Milan, ItalyStem Cells Research Laboratory, Department of Obstetrics & Gynecology, Geneva University, Hospital Geneva, SwitzerlandDepartment of Pharmacological Sciences and Centre for Stem Cell Research, University of Milan, Milan, Italy; Corresponding author at: Department of Pharmacological Sciences and Center for Stem Cell Research, Università degli Studi di Milano, Via Balzaretti, 9, 20133 Milano, Italy. Fax: +39 025025843.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.http://www.sciencedirect.com/science/article/pii/S0969996111004177Human induced pluripotent stem cellsHuntington's diseaseNeuronal differentiation |
| spellingShingle | Stefano Camnasio Alessia Delli Carri Angelo Lombardo Iwona Grad Caterina Mariotti Alessia Castucci Björn Rozell Pietro Lo Riso Valentina Castiglioni Chiara Zuccato Christelle Rochon Yasuhiro Takashima Giuseppe Diaferia Ida Biunno Cinzia Gellera Marisa Jaconi Austin Smith Outi Hovatta Luigi Naldini Stefano Di Donato Anis Feki Elena Cattaneo 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 Neurobiology of Disease Human induced pluripotent stem cells Huntington's disease Neuronal differentiation |
| 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 |
| title_full | 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 |
| title_fullStr | 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 |
| title_full_unstemmed | 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 |
| title_short | 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 |
| title_sort | first reported generation of several induced pluripotent stem cell lines from homozygous and heterozygous huntington s disease patients demonstrates mutation related enhanced lysosomal activity |
| topic | Human induced pluripotent stem cells Huntington's disease Neuronal differentiation |
| url | http://www.sciencedirect.com/science/article/pii/S0969996111004177 |
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