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 <i>HTT</i> gene. Here we report the introduction of an HD mutation into the genome of healthy human embryonic fibroblasts through CRISPR/Cas9-mediated homologous re...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-11-01
|
| Series: | Journal of Personalized Medicine |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2075-4426/10/4/215 |
| _version_ | 1797548495326937088 |
|---|---|
| author | Tuyana Malankhanova Lyubov Suldina Elena Grigor’eva Sergey Medvedev Julia Minina Ksenia Morozova Elena Kiseleva Suren Zakian Anastasia Malakhova |
| author_facet | Tuyana Malankhanova Lyubov Suldina Elena Grigor’eva Sergey Medvedev Julia Minina Ksenia Morozova Elena Kiseleva Suren Zakian Anastasia Malakhova |
| author_sort | Tuyana Malankhanova |
| collection | DOAJ |
| description | Huntington’s disease (HD) is a severe neurodegenerative disorder caused by a CAG triplet expansion in the first exon of the <i>HTT</i> 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 <i>HTT</i>-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. |
| first_indexed | 2024-03-10T15:00:11Z |
| format | Article |
| id | doaj.art-cd8f329414bf4320becf11c0bde0563e |
| institution | Directory Open Access Journal |
| issn | 2075-4426 |
| language | English |
| last_indexed | 2024-03-10T15:00:11Z |
| publishDate | 2020-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Personalized Medicine |
| spelling | doaj.art-cd8f329414bf4320becf11c0bde0563e2023-11-20T20:15:12ZengMDPI AGJournal of Personalized Medicine2075-44262020-11-0110421510.3390/jpm10040215A Human Induced Pluripotent Stem Cell-Derived Isogenic Model of Huntington’s Disease Based on Neuronal Cells Has Several Relevant Phenotypic AbnormalitiesTuyana Malankhanova0Lyubov Suldina1Elena Grigor’eva2Sergey Medvedev3Julia Minina4Ksenia Morozova5Elena Kiseleva6Suren Zakian7Anastasia Malakhova8Federal Research Center Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences, 630090 Novosibirsk, RussiaFederal Research Center Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences, 630090 Novosibirsk, RussiaFederal Research Center Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences, 630090 Novosibirsk, RussiaFederal Research Center Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences, 630090 Novosibirsk, RussiaFederal Research Center Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences, 630090 Novosibirsk, RussiaFederal Research Center Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences, 630090 Novosibirsk, RussiaFederal Research Center Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences, 630090 Novosibirsk, RussiaFederal Research Center Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences, 630090 Novosibirsk, RussiaFederal Research Center Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences, 630090 Novosibirsk, RussiaHuntington’s disease (HD) is a severe neurodegenerative disorder caused by a CAG triplet expansion in the first exon of the <i>HTT</i> 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 <i>HTT</i>-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.https://www.mdpi.com/2075-4426/10/4/215Huntington’s disease modelinggenome editinginduced pluripotent stem cells |
| spellingShingle | Tuyana Malankhanova Lyubov Suldina Elena Grigor’eva Sergey Medvedev Julia Minina Ksenia Morozova Elena Kiseleva Suren Zakian Anastasia Malakhova A Human Induced Pluripotent Stem Cell-Derived Isogenic Model of Huntington’s Disease Based on Neuronal Cells Has Several Relevant Phenotypic Abnormalities Journal of Personalized Medicine Huntington’s disease modeling genome editing induced pluripotent stem cells |
| 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 | Huntington’s disease modeling genome editing induced pluripotent stem cells |
| url | https://www.mdpi.com/2075-4426/10/4/215 |
| work_keys_str_mv | AT tuyanamalankhanova ahumaninducedpluripotentstemcellderivedisogenicmodelofhuntingtonsdiseasebasedonneuronalcellshasseveralrelevantphenotypicabnormalities AT lyubovsuldina ahumaninducedpluripotentstemcellderivedisogenicmodelofhuntingtonsdiseasebasedonneuronalcellshasseveralrelevantphenotypicabnormalities AT elenagrigoreva ahumaninducedpluripotentstemcellderivedisogenicmodelofhuntingtonsdiseasebasedonneuronalcellshasseveralrelevantphenotypicabnormalities AT sergeymedvedev ahumaninducedpluripotentstemcellderivedisogenicmodelofhuntingtonsdiseasebasedonneuronalcellshasseveralrelevantphenotypicabnormalities AT juliaminina ahumaninducedpluripotentstemcellderivedisogenicmodelofhuntingtonsdiseasebasedonneuronalcellshasseveralrelevantphenotypicabnormalities AT kseniamorozova ahumaninducedpluripotentstemcellderivedisogenicmodelofhuntingtonsdiseasebasedonneuronalcellshasseveralrelevantphenotypicabnormalities AT elenakiseleva ahumaninducedpluripotentstemcellderivedisogenicmodelofhuntingtonsdiseasebasedonneuronalcellshasseveralrelevantphenotypicabnormalities AT surenzakian ahumaninducedpluripotentstemcellderivedisogenicmodelofhuntingtonsdiseasebasedonneuronalcellshasseveralrelevantphenotypicabnormalities AT anastasiamalakhova ahumaninducedpluripotentstemcellderivedisogenicmodelofhuntingtonsdiseasebasedonneuronalcellshasseveralrelevantphenotypicabnormalities AT tuyanamalankhanova humaninducedpluripotentstemcellderivedisogenicmodelofhuntingtonsdiseasebasedonneuronalcellshasseveralrelevantphenotypicabnormalities AT lyubovsuldina humaninducedpluripotentstemcellderivedisogenicmodelofhuntingtonsdiseasebasedonneuronalcellshasseveralrelevantphenotypicabnormalities AT elenagrigoreva humaninducedpluripotentstemcellderivedisogenicmodelofhuntingtonsdiseasebasedonneuronalcellshasseveralrelevantphenotypicabnormalities AT sergeymedvedev humaninducedpluripotentstemcellderivedisogenicmodelofhuntingtonsdiseasebasedonneuronalcellshasseveralrelevantphenotypicabnormalities AT juliaminina humaninducedpluripotentstemcellderivedisogenicmodelofhuntingtonsdiseasebasedonneuronalcellshasseveralrelevantphenotypicabnormalities AT kseniamorozova humaninducedpluripotentstemcellderivedisogenicmodelofhuntingtonsdiseasebasedonneuronalcellshasseveralrelevantphenotypicabnormalities AT elenakiseleva humaninducedpluripotentstemcellderivedisogenicmodelofhuntingtonsdiseasebasedonneuronalcellshasseveralrelevantphenotypicabnormalities AT surenzakian humaninducedpluripotentstemcellderivedisogenicmodelofhuntingtonsdiseasebasedonneuronalcellshasseveralrelevantphenotypicabnormalities AT anastasiamalakhova humaninducedpluripotentstemcellderivedisogenicmodelofhuntingtonsdiseasebasedonneuronalcellshasseveralrelevantphenotypicabnormalities |