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A Human Induced Pluripotent Stem Cell-Derived Isogenic Model of Huntington’s Disease Based on Neuronal Cells Has Several Relevant Phenotypic Abnormalities

Huntington’s disease (HD) is a severe neurodegenerative disorder caused by a CAG triplet expansion in the first exon of the HTT gene. Here we report the introduction of an HD mutation into the genome of healthy human embryonic fibroblasts through CRISPR/Cas9-mediated homologous recombination. We ver...

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Autores principales: Malankhanova, Tuyana, Suldina, Lyubov, Grigor’eva, Elena, Medvedev, Sergey, Minina, Julia, Morozova, Ksenia, Kiseleva, Elena, Zakian, Suren, Malakhova, Anastasia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7712151/
https://www.ncbi.nlm.nih.gov/pubmed/33182269
http://dx.doi.org/10.3390/jpm10040215
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author Malankhanova, Tuyana
Suldina, Lyubov
Grigor’eva, Elena
Medvedev, Sergey
Minina, Julia
Morozova, Ksenia
Kiseleva, Elena
Zakian, Suren
Malakhova, Anastasia
author_facet Malankhanova, Tuyana
Suldina, Lyubov
Grigor’eva, Elena
Medvedev, Sergey
Minina, Julia
Morozova, Ksenia
Kiseleva, Elena
Zakian, Suren
Malakhova, Anastasia
author_sort Malankhanova, Tuyana
collection PubMed
description Huntington’s disease (HD) is a severe neurodegenerative disorder caused by a CAG triplet expansion in the first exon of the HTT gene. Here we report the introduction of an HD mutation into the genome of healthy human embryonic fibroblasts through CRISPR/Cas9-mediated homologous recombination. We verified the specificity of the created HTT-editing system and confirmed the absence of undesirable genomic modifications at off-target sites. We showed that both mutant and control isogenic induced pluripotent stem cells (iPSCs) derived by reprogramming of the fibroblast clones can be differentiated into striatal medium spiny neurons. We next demonstrated phenotypic abnormalities in the mutant iPSC-derived neural cells, including impaired neural rosette formation and increased sensitivity to growth factor withdrawal. Moreover, using electron microscopic analysis, we detected a series of ultrastructural defects in the mutant neurons, which did not contain huntingtin aggregates, suggesting that these defects appear early in HD development. Thus, our study describes creation of a new isogenic iPSC-based cell system that models HD and recapitulates HD-specific disturbances in the mutant cells, including some ultrastructural features implemented for the first time.
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spelling pubmed-77121512020-12-04 A Human Induced Pluripotent Stem Cell-Derived Isogenic Model of Huntington’s Disease Based on Neuronal Cells Has Several Relevant Phenotypic Abnormalities Malankhanova, Tuyana Suldina, Lyubov Grigor’eva, Elena Medvedev, Sergey Minina, Julia Morozova, Ksenia Kiseleva, Elena Zakian, Suren Malakhova, Anastasia J Pers Med Article Huntington’s disease (HD) is a severe neurodegenerative disorder caused by a CAG triplet expansion in the first exon of the HTT gene. Here we report the introduction of an HD mutation into the genome of healthy human embryonic fibroblasts through CRISPR/Cas9-mediated homologous recombination. We verified the specificity of the created HTT-editing system and confirmed the absence of undesirable genomic modifications at off-target sites. We showed that both mutant and control isogenic induced pluripotent stem cells (iPSCs) derived by reprogramming of the fibroblast clones can be differentiated into striatal medium spiny neurons. We next demonstrated phenotypic abnormalities in the mutant iPSC-derived neural cells, including impaired neural rosette formation and increased sensitivity to growth factor withdrawal. Moreover, using electron microscopic analysis, we detected a series of ultrastructural defects in the mutant neurons, which did not contain huntingtin aggregates, suggesting that these defects appear early in HD development. Thus, our study describes creation of a new isogenic iPSC-based cell system that models HD and recapitulates HD-specific disturbances in the mutant cells, including some ultrastructural features implemented for the first time. MDPI 2020-11-09 /pmc/articles/PMC7712151/ /pubmed/33182269 http://dx.doi.org/10.3390/jpm10040215 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Malankhanova, Tuyana
Suldina, Lyubov
Grigor’eva, Elena
Medvedev, Sergey
Minina, Julia
Morozova, Ksenia
Kiseleva, Elena
Zakian, Suren
Malakhova, Anastasia
A Human Induced Pluripotent Stem Cell-Derived Isogenic Model of Huntington’s Disease Based on Neuronal Cells Has Several Relevant Phenotypic Abnormalities
title A Human Induced Pluripotent Stem Cell-Derived Isogenic Model of Huntington’s Disease Based on Neuronal Cells Has Several Relevant Phenotypic Abnormalities
title_full A Human Induced Pluripotent Stem Cell-Derived Isogenic Model of Huntington’s Disease Based on Neuronal Cells Has Several Relevant Phenotypic Abnormalities
title_fullStr A Human Induced Pluripotent Stem Cell-Derived Isogenic Model of Huntington’s Disease Based on Neuronal Cells Has Several Relevant Phenotypic Abnormalities
title_full_unstemmed A Human Induced Pluripotent Stem Cell-Derived Isogenic Model of Huntington’s Disease Based on Neuronal Cells Has Several Relevant Phenotypic Abnormalities
title_short A Human Induced Pluripotent Stem Cell-Derived Isogenic Model of Huntington’s Disease Based on Neuronal Cells Has Several Relevant Phenotypic Abnormalities
title_sort human induced pluripotent stem cell-derived isogenic model of huntington’s disease based on neuronal cells has several relevant phenotypic abnormalities
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7712151/
https://www.ncbi.nlm.nih.gov/pubmed/33182269
http://dx.doi.org/10.3390/jpm10040215
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