Allele-Specific Gene Editing Rescues Pathology in a Human Model of Charcot-Marie-Tooth Disease Type 2E
Many neuromuscular disorders are caused by dominant missense mutations that lead to dominant-negative or gain-of-function pathology. This category of disease is challenging to address via drug treatment or gene augmentation therapy because these strategies may not eliminate the effects of the mutant...
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| Format: | Article |
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Frontiers Media S.A.
2021-08-01
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| Series: | Frontiers in Cell and Developmental Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2021.723023/full |
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| author | Carissa M. Feliciano Carissa M. Feliciano Kenneth Wu Hannah L. Watry Chiara B. E. Marley Chiara B. E. Marley Gokul N. Ramadoss Gokul N. Ramadoss Hana Y. Ghanim Angela Z. Liu Angela Z. Liu Lyandysha V. Zholudeva Todd C. McDevitt Todd C. McDevitt Mario A. Saporta Bruce R. Conklin Bruce R. Conklin Bruce R. Conklin Bruce R. Conklin Luke M. Judge Luke M. Judge |
| author_facet | Carissa M. Feliciano Carissa M. Feliciano Kenneth Wu Hannah L. Watry Chiara B. E. Marley Chiara B. E. Marley Gokul N. Ramadoss Gokul N. Ramadoss Hana Y. Ghanim Angela Z. Liu Angela Z. Liu Lyandysha V. Zholudeva Todd C. McDevitt Todd C. McDevitt Mario A. Saporta Bruce R. Conklin Bruce R. Conklin Bruce R. Conklin Bruce R. Conklin Luke M. Judge Luke M. Judge |
| author_sort | Carissa M. Feliciano |
| collection | DOAJ |
| description | Many neuromuscular disorders are caused by dominant missense mutations that lead to dominant-negative or gain-of-function pathology. This category of disease is challenging to address via drug treatment or gene augmentation therapy because these strategies may not eliminate the effects of the mutant protein or RNA. Thus, effective treatments are severely lacking for these dominant diseases, which often cause severe disability or death. The targeted inactivation of dominant disease alleles by gene editing is a promising approach with the potential to completely remove the cause of pathology with a single treatment. Here, we demonstrate that allele-specific CRISPR gene editing in a human model of axonal Charcot-Marie-Tooth (CMT) disease rescues pathology caused by a dominant missense mutation in the neurofilament light chain gene (NEFL, CMT type 2E). We utilized a rapid and efficient method for generating spinal motor neurons from human induced pluripotent stem cells (iPSCs) derived from a patient with CMT2E. Diseased motor neurons recapitulated known pathologic phenotypes at early time points of differentiation, including aberrant accumulation of neurofilament light chain protein in neuronal cell bodies. We selectively inactivated the disease NEFL allele in patient iPSCs using Cas9 enzymes to introduce a frameshift at the pathogenic N98S mutation. Motor neurons carrying this allele-specific frameshift demonstrated an amelioration of the disease phenotype comparable to that seen in an isogenic control with precise correction of the mutation. Our results validate allele-specific gene editing as a therapeutic approach for CMT2E and as a promising strategy to silence dominant mutations in any gene for which heterozygous loss-of-function is well tolerated. This highlights the potential for gene editing as a therapy for currently untreatable dominant neurologic diseases. |
| first_indexed | 2024-12-16T11:53:40Z |
| format | Article |
| id | doaj.art-1c701f1854d94bbf83e09940f948c98f |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-12-16T11:53:40Z |
| publishDate | 2021-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-1c701f1854d94bbf83e09940f948c98f2022-12-21T22:32:37ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2021-08-01910.3389/fcell.2021.723023723023Allele-Specific Gene Editing Rescues Pathology in a Human Model of Charcot-Marie-Tooth Disease Type 2ECarissa M. Feliciano0Carissa M. Feliciano1Kenneth Wu2Hannah L. Watry3Chiara B. E. Marley4Chiara B. E. Marley5Gokul N. Ramadoss6Gokul N. Ramadoss7Hana Y. Ghanim8Angela Z. Liu9Angela Z. Liu10Lyandysha V. Zholudeva11Todd C. McDevitt12Todd C. McDevitt13Mario A. Saporta14Bruce R. Conklin15Bruce R. Conklin16Bruce R. Conklin17Bruce R. Conklin18Luke M. Judge19Luke M. Judge20Department of Pediatrics, University of California, San Francisco, San Francisco, CA, United StatesGladstone Institutes, San Francisco, CA, United StatesGladstone Institutes, San Francisco, CA, United StatesGladstone Institutes, San Francisco, CA, United StatesDepartment of Pediatrics, University of California, San Francisco, San Francisco, CA, United StatesGladstone Institutes, San Francisco, CA, United StatesGladstone Institutes, San Francisco, CA, United StatesBiomedical Sciences Ph.D. Program, University of California, San Francisco, San Francisco, CA, United StatesGladstone Institutes, San Francisco, CA, United StatesGladstone Institutes, San Francisco, CA, United StatesDepartment of Ophthalmology, University of California, San Francisco, San Francisco, CA, United StatesGladstone Institutes, San Francisco, CA, United StatesGladstone Institutes, San Francisco, CA, United StatesDepartment of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, CA, United StatesDepartment of Neurology, Miller School of Medicine, University of Miami, Miami, FL, United StatesGladstone Institutes, San Francisco, CA, United StatesDepartment of Ophthalmology, University of California, San Francisco, San Francisco, CA, United StatesDepartment of Medicine, University of California, San Francisco, San Francisco, CA, United StatesInnovative Genomics Institute, Berkeley, CA, United StatesDepartment of Pediatrics, University of California, San Francisco, San Francisco, CA, United StatesGladstone Institutes, San Francisco, CA, United StatesMany neuromuscular disorders are caused by dominant missense mutations that lead to dominant-negative or gain-of-function pathology. This category of disease is challenging to address via drug treatment or gene augmentation therapy because these strategies may not eliminate the effects of the mutant protein or RNA. Thus, effective treatments are severely lacking for these dominant diseases, which often cause severe disability or death. The targeted inactivation of dominant disease alleles by gene editing is a promising approach with the potential to completely remove the cause of pathology with a single treatment. Here, we demonstrate that allele-specific CRISPR gene editing in a human model of axonal Charcot-Marie-Tooth (CMT) disease rescues pathology caused by a dominant missense mutation in the neurofilament light chain gene (NEFL, CMT type 2E). We utilized a rapid and efficient method for generating spinal motor neurons from human induced pluripotent stem cells (iPSCs) derived from a patient with CMT2E. Diseased motor neurons recapitulated known pathologic phenotypes at early time points of differentiation, including aberrant accumulation of neurofilament light chain protein in neuronal cell bodies. We selectively inactivated the disease NEFL allele in patient iPSCs using Cas9 enzymes to introduce a frameshift at the pathogenic N98S mutation. Motor neurons carrying this allele-specific frameshift demonstrated an amelioration of the disease phenotype comparable to that seen in an isogenic control with precise correction of the mutation. Our results validate allele-specific gene editing as a therapeutic approach for CMT2E and as a promising strategy to silence dominant mutations in any gene for which heterozygous loss-of-function is well tolerated. This highlights the potential for gene editing as a therapy for currently untreatable dominant neurologic diseases.https://www.frontiersin.org/articles/10.3389/fcell.2021.723023/fullinduced pluripotent stem cellsCharcot-Marie-Toothneuropathymotor neuronsdominantCRISPR-Cas9 |
| spellingShingle | Carissa M. Feliciano Carissa M. Feliciano Kenneth Wu Hannah L. Watry Chiara B. E. Marley Chiara B. E. Marley Gokul N. Ramadoss Gokul N. Ramadoss Hana Y. Ghanim Angela Z. Liu Angela Z. Liu Lyandysha V. Zholudeva Todd C. McDevitt Todd C. McDevitt Mario A. Saporta Bruce R. Conklin Bruce R. Conklin Bruce R. Conklin Bruce R. Conklin Luke M. Judge Luke M. Judge Allele-Specific Gene Editing Rescues Pathology in a Human Model of Charcot-Marie-Tooth Disease Type 2E Frontiers in Cell and Developmental Biology induced pluripotent stem cells Charcot-Marie-Tooth neuropathy motor neurons dominant CRISPR-Cas9 |
| title | Allele-Specific Gene Editing Rescues Pathology in a Human Model of Charcot-Marie-Tooth Disease Type 2E |
| title_full | Allele-Specific Gene Editing Rescues Pathology in a Human Model of Charcot-Marie-Tooth Disease Type 2E |
| title_fullStr | Allele-Specific Gene Editing Rescues Pathology in a Human Model of Charcot-Marie-Tooth Disease Type 2E |
| title_full_unstemmed | Allele-Specific Gene Editing Rescues Pathology in a Human Model of Charcot-Marie-Tooth Disease Type 2E |
| title_short | Allele-Specific Gene Editing Rescues Pathology in a Human Model of Charcot-Marie-Tooth Disease Type 2E |
| title_sort | allele specific gene editing rescues pathology in a human model of charcot marie tooth disease type 2e |
| topic | induced pluripotent stem cells Charcot-Marie-Tooth neuropathy motor neurons dominant CRISPR-Cas9 |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2021.723023/full |
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