Genome editing using Staphylococcus aureus Cas9 in a canine model of glycogen storage disease Ia
Glycogen storage disease type Ia (GSD Ia) is the inherited deficiency of glucose-6-phosphatase (G6Pase), associated with life-threatening hypoglycemia and long-term complications, including hepatocellular carcinoma formation. Gene replacement therapy fails to stably reverse G6Pase deficiency. We att...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-06-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050123000372 |
| _version_ | 1797807184255385600 |
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| author | Benjamin Arnson Hye Ri Kang Elizabeth D. Brooks Dorothy Gheorghiu Ekaterina Ilich David Courtney Jeffrey I. Everitt Bryan R. Cullen Dwight D. Koeberl |
| author_facet | Benjamin Arnson Hye Ri Kang Elizabeth D. Brooks Dorothy Gheorghiu Ekaterina Ilich David Courtney Jeffrey I. Everitt Bryan R. Cullen Dwight D. Koeberl |
| author_sort | Benjamin Arnson |
| collection | DOAJ |
| description | Glycogen storage disease type Ia (GSD Ia) is the inherited deficiency of glucose-6-phosphatase (G6Pase), associated with life-threatening hypoglycemia and long-term complications, including hepatocellular carcinoma formation. Gene replacement therapy fails to stably reverse G6Pase deficiency. We attempted genome editing using two adeno-associated virus vectors, one that expressed Staphylococcus aureus Cas9 protein and a second containing a donor transgene encoding G6Pase, in a dog model for GSD Ia. We demonstrated donor transgene integration in the liver of three adult-treated dogs accompanied by stable G6Pase expression and correction of hypoglycemia during fasting. Two puppies with GSD Ia were treated by genome editing that achieved donor transgene integration in the liver. Integration frequency ranged from 0.5% to 1% for all dogs. In adult-treated dogs, anti-SaCas9 antibodies were detected before genome editing, reflecting prior exposure to S. aureus. Nuclease activity was low, as reflected by a low percentage of indel formation at the predicted site of SaCas9 cutting that indicated double-stranded breaks followed by non-homologous end-joining. Thus, genome editing can integrate a therapeutic transgene in the liver of a large animal model, either early or later in life, and further development is warranted to provide a more stable treatment for GSD Ia. |
| first_indexed | 2024-03-13T06:18:45Z |
| format | Article |
| id | doaj.art-8282e120c0924cec897cf976a2665255 |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-03-13T06:18:45Z |
| publishDate | 2023-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-8282e120c0924cec897cf976a26652552023-06-10T04:27:42ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012023-06-0129108119Genome editing using Staphylococcus aureus Cas9 in a canine model of glycogen storage disease IaBenjamin Arnson0Hye Ri Kang1Elizabeth D. Brooks2Dorothy Gheorghiu3Ekaterina Ilich4David Courtney5Jeffrey I. Everitt6Bryan R. Cullen7Dwight D. Koeberl8Division of Medical Genetics, Department of Pediatrics, Duke University Medical School, Durham, NC, USA; Department of Molecular Genetics & Microbiology, Duke University Medical Center, Durham, NC, USADepartment of Pediatrics, UT Southwestern Medical Center, Dallas, TX, USADivision of Medical Genetics, Department of Pediatrics, Duke University Medical School, Durham, NC, USADivision of Medical Genetics, Department of Pediatrics, Duke University Medical School, Durham, NC, USADivision of Medical Genetics, Department of Pediatrics, Duke University Medical School, Durham, NC, USAWellcome-Wolfson Institute for Experimental Medicine, Queen’s University Belfast, Belfast, UKDepartment of Pathology, Duke University School of Medicine, Durham, NC, USADepartment of Molecular Genetics & Microbiology, Duke University Medical Center, Durham, NC, USADivision of Medical Genetics, Department of Pediatrics, Duke University Medical School, Durham, NC, USA; Department of Molecular Genetics & Microbiology, Duke University Medical Center, Durham, NC, USA; Corresponding author: Dwight D. Koeberl, Box 103856, Duke University Medical Center, Durham, NC 27710, USA.Glycogen storage disease type Ia (GSD Ia) is the inherited deficiency of glucose-6-phosphatase (G6Pase), associated with life-threatening hypoglycemia and long-term complications, including hepatocellular carcinoma formation. Gene replacement therapy fails to stably reverse G6Pase deficiency. We attempted genome editing using two adeno-associated virus vectors, one that expressed Staphylococcus aureus Cas9 protein and a second containing a donor transgene encoding G6Pase, in a dog model for GSD Ia. We demonstrated donor transgene integration in the liver of three adult-treated dogs accompanied by stable G6Pase expression and correction of hypoglycemia during fasting. Two puppies with GSD Ia were treated by genome editing that achieved donor transgene integration in the liver. Integration frequency ranged from 0.5% to 1% for all dogs. In adult-treated dogs, anti-SaCas9 antibodies were detected before genome editing, reflecting prior exposure to S. aureus. Nuclease activity was low, as reflected by a low percentage of indel formation at the predicted site of SaCas9 cutting that indicated double-stranded breaks followed by non-homologous end-joining. Thus, genome editing can integrate a therapeutic transgene in the liver of a large animal model, either early or later in life, and further development is warranted to provide a more stable treatment for GSD Ia.http://www.sciencedirect.com/science/article/pii/S2329050123000372genome editingCRISPR-Cas9adeno-associated virus vectorsinherited disorder of metabolismglycogen storage diseaseglucose-6-phosphatase |
| spellingShingle | Benjamin Arnson Hye Ri Kang Elizabeth D. Brooks Dorothy Gheorghiu Ekaterina Ilich David Courtney Jeffrey I. Everitt Bryan R. Cullen Dwight D. Koeberl Genome editing using Staphylococcus aureus Cas9 in a canine model of glycogen storage disease Ia Molecular Therapy: Methods & Clinical Development genome editing CRISPR-Cas9 adeno-associated virus vectors inherited disorder of metabolism glycogen storage disease glucose-6-phosphatase |
| title | Genome editing using Staphylococcus aureus Cas9 in a canine model of glycogen storage disease Ia |
| title_full | Genome editing using Staphylococcus aureus Cas9 in a canine model of glycogen storage disease Ia |
| title_fullStr | Genome editing using Staphylococcus aureus Cas9 in a canine model of glycogen storage disease Ia |
| title_full_unstemmed | Genome editing using Staphylococcus aureus Cas9 in a canine model of glycogen storage disease Ia |
| title_short | Genome editing using Staphylococcus aureus Cas9 in a canine model of glycogen storage disease Ia |
| title_sort | genome editing using staphylococcus aureus cas9 in a canine model of glycogen storage disease ia |
| topic | genome editing CRISPR-Cas9 adeno-associated virus vectors inherited disorder of metabolism glycogen storage disease glucose-6-phosphatase |
| url | http://www.sciencedirect.com/science/article/pii/S2329050123000372 |
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