A comparison of AAV strategies distinguishes overlapping vectors for efficient systemic delivery of the 6.2 kb Dysferlin coding sequence
Recombinant adeno-associated virus (rAAV) is currently the best vector for gene delivery into the skeletal muscle. However, the 5-kb packaging size of this virus is a major obstacle for large gene transfer. This past decade, many different strategies were developed to circumvent this issue (concatem...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2015-01-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050116300213 |
| _version_ | 1818173713716084736 |
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| author | Marina Pryadkina William Lostal Nathalie Bourg Karine Charton Carinne Roudaut Matthew L Hirsch Isabelle Richard |
| author_facet | Marina Pryadkina William Lostal Nathalie Bourg Karine Charton Carinne Roudaut Matthew L Hirsch Isabelle Richard |
| author_sort | Marina Pryadkina |
| collection | DOAJ |
| description | Recombinant adeno-associated virus (rAAV) is currently the best vector for gene delivery into the skeletal muscle. However, the 5-kb packaging size of this virus is a major obstacle for large gene transfer. This past decade, many different strategies were developed to circumvent this issue (concatemerization-splicing, overlapping vectors, hybrid dual or fragmented AAV). Loss of function mutations in the DYSF gene whose coding sequence is 6.2kb lead to progressive muscular dystrophies (LGMD2B: OMIM_253601; MM: OMIM_254130; DMAT: OMIM_606768). In this study, we compared large gene transfer techniques to deliver the DYSF gene into the skeletal muscle. After rAAV8s intramuscular injection into dysferlin deficient mice, we showed that the overlap strategy is the most effective approach to reconstitute a full-length messenger. After systemic administration, the level of dysferlin obtained on different muscles corresponded to 0.5- to 2-fold compared to the normal level. We further demonstrated that the overlapping vector set was efficient to correct the histopathology, resistance to eccentric contractions and whole body force in the dysferlin deficient mice. Altogether, these data indicate that using overlapping vectors could be a promising approach for a potential clinical treatment of dysferlinopathies. |
| first_indexed | 2024-12-11T19:32:53Z |
| format | Article |
| id | doaj.art-8276dca3a8bc48b48b6464d10a8a1df1 |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-12-11T19:32:53Z |
| publishDate | 2015-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-8276dca3a8bc48b48b6464d10a8a1df12022-12-22T00:53:14ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012015-01-012A comparison of AAV strategies distinguishes overlapping vectors for efficient systemic delivery of the 6.2 kb Dysferlin coding sequenceMarina Pryadkina0William Lostal1Nathalie Bourg2Karine Charton3Carinne Roudaut4Matthew L Hirsch5Isabelle Richard6Inserm, U951, Evry, France; Genethon, R&D Department, INTEGRARE Research Unit, Evry, FranceInserm, U951, Evry, France; Genethon, R&D Department, INTEGRARE Research Unit, Evry, FranceInserm, U951, Evry, France; Genethon, R&D Department, INTEGRARE Research Unit, Evry, FranceInserm, U951, Evry, France; Genethon, R&D Department, INTEGRARE Research Unit, Evry, FranceInserm, U951, Evry, France; Genethon, R&D Department, INTEGRARE Research Unit, Evry, FranceGene Therapy Center, University of North Carolina, Chapel Hill, North Carolina, USA; Department of Ophthalmology, University of North Carolina, Chapel Hill, North Carolina, USAInserm, U951, Evry, France; Genethon, R&D Department, INTEGRARE Research Unit, Evry, FranceRecombinant adeno-associated virus (rAAV) is currently the best vector for gene delivery into the skeletal muscle. However, the 5-kb packaging size of this virus is a major obstacle for large gene transfer. This past decade, many different strategies were developed to circumvent this issue (concatemerization-splicing, overlapping vectors, hybrid dual or fragmented AAV). Loss of function mutations in the DYSF gene whose coding sequence is 6.2kb lead to progressive muscular dystrophies (LGMD2B: OMIM_253601; MM: OMIM_254130; DMAT: OMIM_606768). In this study, we compared large gene transfer techniques to deliver the DYSF gene into the skeletal muscle. After rAAV8s intramuscular injection into dysferlin deficient mice, we showed that the overlap strategy is the most effective approach to reconstitute a full-length messenger. After systemic administration, the level of dysferlin obtained on different muscles corresponded to 0.5- to 2-fold compared to the normal level. We further demonstrated that the overlapping vector set was efficient to correct the histopathology, resistance to eccentric contractions and whole body force in the dysferlin deficient mice. Altogether, these data indicate that using overlapping vectors could be a promising approach for a potential clinical treatment of dysferlinopathies.http://www.sciencedirect.com/science/article/pii/S2329050116300213 |
| spellingShingle | Marina Pryadkina William Lostal Nathalie Bourg Karine Charton Carinne Roudaut Matthew L Hirsch Isabelle Richard A comparison of AAV strategies distinguishes overlapping vectors for efficient systemic delivery of the 6.2 kb Dysferlin coding sequence Molecular Therapy: Methods & Clinical Development |
| title | A comparison of AAV strategies distinguishes overlapping vectors for efficient systemic delivery of the 6.2 kb Dysferlin coding sequence |
| title_full | A comparison of AAV strategies distinguishes overlapping vectors for efficient systemic delivery of the 6.2 kb Dysferlin coding sequence |
| title_fullStr | A comparison of AAV strategies distinguishes overlapping vectors for efficient systemic delivery of the 6.2 kb Dysferlin coding sequence |
| title_full_unstemmed | A comparison of AAV strategies distinguishes overlapping vectors for efficient systemic delivery of the 6.2 kb Dysferlin coding sequence |
| title_short | A comparison of AAV strategies distinguishes overlapping vectors for efficient systemic delivery of the 6.2 kb Dysferlin coding sequence |
| title_sort | comparison of aav strategies distinguishes overlapping vectors for efficient systemic delivery of the 6 2 kb dysferlin coding sequence |
| url | http://www.sciencedirect.com/science/article/pii/S2329050116300213 |
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