AAV CRISPR editing rescues cardiac and muscle function for 18 months in dystrophic mice
Adeno-associated virus-mediated (AAV-mediated) CRISPR editing is a revolutionary approach for treating inherited diseases. Sustained, often life-long mutation correction is required for treating these diseases. Unfortunately, this has never been demonstrated with AAV CRISPR therapy. We addressed thi...
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Published in | JCI insight Vol. 3; no. 23 |
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Main Authors | , , , , , , , , , , , , , , , , |
Format | Journal Article |
Language | English |
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United States
American Society for Clinical Investigation
06.12.2018
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Abstract | Adeno-associated virus-mediated (AAV-mediated) CRISPR editing is a revolutionary approach for treating inherited diseases. Sustained, often life-long mutation correction is required for treating these diseases. Unfortunately, this has never been demonstrated with AAV CRISPR therapy. We addressed this question in the mdx model of Duchenne muscular dystrophy (DMD). DMD is caused by dystrophin gene mutation. Dystrophin deficiency leads to ambulation loss and cardiomyopathy. We treated 6-week-old mice intravenously and evaluated disease rescue at 18 months. Surprisingly, nominal dystrophin was restored in skeletal muscle. Cardiac dystrophin was restored, but histology and hemodynamics were not improved. To determine the underlying mechanism, we evaluated components of the CRISPR-editing machinery. Intriguingly, we found disproportional guide RNA (gRNA) vector depletion. To test whether this is responsible for the poor outcome, we increased the gRNA vector dose and repeated the study. This strategy significantly increased dystrophin restoration and reduced fibrosis in all striated muscles at 18 months. Importantly, skeletal muscle function and cardiac hemodynamics were significantly enhanced. Interestingly, we did not see selective depletion of the gRNA vector after intramuscular injection. Our results suggest that gRNA vector loss is a unique barrier for systemic AAV CRISPR therapy. This can be circumvented by vector dose optimization. |
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AbstractList | Adeno-associated virus-mediated (AAV-mediated) CRISPR editing is a revolutionary approach for treating inherited diseases. Sustained, often life-long mutation correction is required for treating these diseases. Unfortunately, this has never been demonstrated with AAV CRISPR therapy. We addressed this question in the mdx model of Duchenne muscular dystrophy (DMD). DMD is caused by dystrophin gene mutation. Dystrophin deficiency leads to ambulation loss and cardiomyopathy. We treated 6-week-old mice intravenously and evaluated disease rescue at 18 months. Surprisingly, nominal dystrophin was restored in skeletal muscle. Cardiac dystrophin was restored, but histology and hemodynamics were not improved. To determine the underlying mechanism, we evaluated components of the CRISPR-editing machinery. Intriguingly, we found disproportional guide RNA (gRNA) vector depletion. To test whether this is responsible for the poor outcome, we increased the gRNA vector dose and repeated the study. This strategy significantly increased dystrophin restoration and reduced fibrosis in all striated muscles at 18 months. Importantly, skeletal muscle function and cardiac hemodynamics were significantly enhanced. Interestingly, we did not see selective depletion of the gRNA vector after intramuscular injection. Our results suggest that gRNA vector loss is a unique barrier for systemic AAV CRISPR therapy. This can be circumvented by vector dose optimization. Adeno-associated virus–mediated (AAV-mediated) CRISPR editing is a revolutionary approach for treating inherited diseases. Sustained, often life-long mutation correction is required for treating these diseases. Unfortunately, this has never been demonstrated with AAV CRISPR therapy. We addressed this question in the mdx model of Duchenne muscular dystrophy (DMD). DMD is caused by dystrophin gene mutation. Dystrophin deficiency leads to ambulation loss and cardiomyopathy. We treated 6-week-old mice intravenously and evaluated disease rescue at 18 months. Surprisingly, nominal dystrophin was restored in skeletal muscle. Cardiac dystrophin was restored, but histology and hemodynamics were not improved. To determine the underlying mechanism, we evaluated components of the CRISPR-editing machinery. Intriguingly, we found disproportional guide RNA (gRNA) vector depletion. To test whether this is responsible for the poor outcome, we increased the gRNA vector dose and repeated the study. This strategy significantly increased dystrophin restoration and reduced fibrosis in all striated muscles at 18 months. Importantly, skeletal muscle function and cardiac hemodynamics were significantly enhanced. Interestingly, we did not see selective depletion of the gRNA vector after intramuscular injection. Our results suggest that gRNA vector loss is a unique barrier for systemic AAV CRISPR therapy. This can be circumvented by vector dose optimization. Vector dose optimization enabled long-lasting cardiac and skeletal muscle rescue in Duchenne muscular dystrophy mice by systemic AAV CRISPR delivery. |
Author | Nance, Michael E Jenkins, Gregory J Lessa, Thais B Yue, Yongping Yang, N Nora Chen, Shi-Jie Hakim, Chady H Gersbach, Charles A Duan, Dongsheng Dai, Aihua Pan, Xiufang Wasala, Lakmini P Nelson, Christopher E Louderman, Jacqueline A Zhang, Keqing Kodippili, Kasun Wasala, Nalinda B |
AuthorAffiliation | 8 Department of Neurology, School of Medicine 4 Center for Genomic and Computational Biology, Duke University, Durham, North Carolina, USA 3 Department of Biomedical Engineering and 10 Department of Biomedical Sciences, College of Veterinary Medicine, University of Missouri, Columbia, Missouri, USA 6 Department of Biochemistry, University of Missouri, Columbia, Missouri, USA 7 Department of Orthopaedic Surgery, Duke University Medical Center, Durham, North Carolina, USA 2 National Center for Advancing Translational Sciences, NIH, Rockville, Maryland, USA 1 Department of Molecular Microbiology and Immunology, School of Medicine, University of Missouri, Columbia, Missouri, USA 5 Department of Physics and 9 Department of Bioengineering, and |
AuthorAffiliation_xml | – name: 5 Department of Physics and – name: 6 Department of Biochemistry, University of Missouri, Columbia, Missouri, USA – name: 8 Department of Neurology, School of Medicine – name: 9 Department of Bioengineering, and – name: 1 Department of Molecular Microbiology and Immunology, School of Medicine, University of Missouri, Columbia, Missouri, USA – name: 7 Department of Orthopaedic Surgery, Duke University Medical Center, Durham, North Carolina, USA – name: 2 National Center for Advancing Translational Sciences, NIH, Rockville, Maryland, USA – name: 4 Center for Genomic and Computational Biology, Duke University, Durham, North Carolina, USA – name: 10 Department of Biomedical Sciences, College of Veterinary Medicine, University of Missouri, Columbia, Missouri, USA – name: 3 Department of Biomedical Engineering and |
Author_xml | – sequence: 1 givenname: Chady H surname: Hakim fullname: Hakim, Chady H organization: National Center for Advancing Translational Sciences, NIH, Rockville, Maryland, USA – sequence: 2 givenname: Nalinda B surname: Wasala fullname: Wasala, Nalinda B organization: Department of Molecular Microbiology and Immunology, School of Medicine, University of Missouri, Columbia, Missouri, USA – sequence: 3 givenname: Christopher E surname: Nelson fullname: Nelson, Christopher E organization: Center for Genomic and Computational Biology, Duke University, Durham, North Carolina, USA – sequence: 4 givenname: Lakmini P surname: Wasala fullname: Wasala, Lakmini P organization: Department of Molecular Microbiology and Immunology, School of Medicine, University of Missouri, Columbia, Missouri, USA – sequence: 5 givenname: Yongping surname: Yue fullname: Yue, Yongping organization: Department of Molecular Microbiology and Immunology, School of Medicine, University of Missouri, Columbia, Missouri, USA – sequence: 6 givenname: Jacqueline A surname: Louderman fullname: Louderman, Jacqueline A organization: Department of Molecular Microbiology and Immunology, School of Medicine, University of Missouri, Columbia, Missouri, USA – sequence: 7 givenname: Thais B surname: Lessa fullname: Lessa, Thais B organization: Department of Molecular Microbiology and Immunology, School of Medicine, University of Missouri, Columbia, Missouri, USA – sequence: 8 givenname: Aihua surname: Dai fullname: Dai, Aihua organization: Department of Molecular Microbiology and Immunology, School of Medicine, University of Missouri, Columbia, Missouri, USA – sequence: 9 givenname: Keqing surname: Zhang fullname: Zhang, Keqing organization: Department of Molecular Microbiology and Immunology, School of Medicine, University of Missouri, Columbia, Missouri, USA – sequence: 10 givenname: Gregory J surname: Jenkins fullname: Jenkins, Gregory J organization: Department of Molecular Microbiology and Immunology, School of Medicine, University of Missouri, Columbia, Missouri, USA – sequence: 11 givenname: Michael E surname: Nance fullname: Nance, Michael E organization: Department of Molecular Microbiology and Immunology, School of Medicine, University of Missouri, Columbia, Missouri, USA – sequence: 12 givenname: Xiufang surname: Pan fullname: Pan, Xiufang organization: Department of Molecular Microbiology and Immunology, School of Medicine, University of Missouri, Columbia, Missouri, USA – sequence: 13 givenname: Kasun surname: Kodippili fullname: Kodippili, Kasun organization: Department of Molecular Microbiology and Immunology, School of Medicine, University of Missouri, Columbia, Missouri, USA – sequence: 14 givenname: N Nora surname: Yang fullname: Yang, N Nora organization: National Center for Advancing Translational Sciences, NIH, Rockville, Maryland, USA – sequence: 15 givenname: Shi-Jie surname: Chen fullname: Chen, Shi-Jie organization: Department of Biochemistry, University of Missouri, Columbia, Missouri, USA – sequence: 16 givenname: Charles A surname: Gersbach fullname: Gersbach, Charles A organization: Department of Orthopaedic Surgery, Duke University Medical Center, Durham, North Carolina, USA – sequence: 17 givenname: Dongsheng surname: Duan fullname: Duan, Dongsheng organization: Department of Biomedical Sciences, College of Veterinary Medicine, University of Missouri, Columbia, Missouri, USA |
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Snippet | Adeno-associated virus-mediated (AAV-mediated) CRISPR editing is a revolutionary approach for treating inherited diseases. Sustained, often life-long mutation... Adeno-associated virus–mediated (AAV-mediated) CRISPR editing is a revolutionary approach for treating inherited diseases. Sustained, often life-long mutation... |
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SubjectTerms | Animals Clustered Regularly Interspaced Short Palindromic Repeats Dependovirus Disease Models, Animal Dystrophin - genetics Female Fibrosis Gene Editing Genetic Therapy Genetic Vectors Male Mice Mice, Inbred mdx Muscle, Skeletal - metabolism Muscle, Skeletal - pathology Muscular Dystrophy, Duchenne - genetics Muscular Dystrophy, Duchenne - pathology Mutation Myocardium - metabolism Myocardium - pathology Neuromuscular Diseases RNA, Guide, CRISPR-Cas Systems |
Title | AAV CRISPR editing rescues cardiac and muscle function for 18 months in dystrophic mice |
URI | https://www.ncbi.nlm.nih.gov/pubmed/30518686 https://search.proquest.com/docview/2158242003 https://pubmed.ncbi.nlm.nih.gov/PMC6328021 |
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