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Cited by 53Virus-based gene therapy researchNeurogenetic and Muscular Disorders ResearchRNA Interference and Gene Delivery

A comparison of AAV strategies distinguishes overlapping vectors for efficient systemic delivery of the 6.2 kb Dysferlin coding sequence

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Pryadkina, Marina, Lostal, William, Bourg, Nathalie, Charton, Karine, Roudaut, Carinne, Hirsch, Matthew L, Richard, Isabelle·2015·Molecular Therapy - Methods & Clinical Development
DOI 10.1038/mtm.2015.9PMC PMC4445010
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Abstract

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.

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  • Overexpression of dysferlin by intramuscular injection of AAV8 encoding dysferlin from either a single oversized vector or two overlapping vectors showed approximately 10-fold higher expression of dysferlin with the dual-vector system.

    “overexpression of dysferlin by intramuscular injection of AAV8 encoding dysferlin from either a single oversized vector or from two overlapping vectors showed a * 10-fold higher expression of dysferlin with the dual-vector system. 45”

    From Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids (2016) · cites this paper as [45]

    The Results section on intramuscular injection reports higher dysferlin expression with the overlapping dual-vector Set 3 than with the oversized fAAV vector. It gives the increase as approximately 4–10-fold relative to Batch 2 fAAV, so the citing sentence’s approximately 10-fold figure omits the lower end of the reported range. (LLM, conf=0.87)

    Experimental — accuracy not yet measured

  • Overlapping vectors work particularly well in muscle tissue, ameliorating phenotypes linked to Duchenne muscular dystrophy and dysferlinopathies.

    “This strategy was shown to work particularly well in muscle tissue, ameliorating phenotypes linked to Duchenne muscular dystrophy (DMD) [43][44][45] and dysferlinopathies [46][47][48].”

    From Circumventing the packaging limit of AAV-mediated gene replacement therapy for neurological disorders (2022) · cites this paper as [47]

  • The nature of the transgene and tissue/cell type can influence the efficiency of dual vectors.

    “it has been reported that the nature of the transgene 47 and tissue/cell type (possibly because of differences in DNA repair pathways) 45,46 can influence the efficiency of dual vectors.”

    From Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids (2016) · cites this paper as [45]

  • Hybrid systems are not always the most efficient vector systems available.

    “It needs to be pointed out, however, that hybrid systems are not always the most efficient vector systems available.”

    From Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids (2016) · cites this paper as [45]

  • Pryadkina et al. obtained similar results when they analyzed the efficiency of dysferlin overexpression obtainable with dual overlapping vector or hybrid vector systems.

    “For instance, Zhang and Duan demonstrated that a simple, dual overlap vector system outperformed three independent hybrid systems, 68 and Pryadkina et al. obtained similar results when they analyzed the efficiency of dysferlin overexpression obtainable with dual overlapping vector or hybrid vector systems. 45”

    From Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids (2016) · cites this paper as [45]

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Authors
Pryadkina, M., Lostal, W., Bourg, N., Charton, K., Roudaut, C., Hirsch, M. L., & Richard, I.
Affiliations
  • Inserm (FR)
  • University of North Carolina at Chapel Hill (US)
Corresponding author
Richard, Isabelle
ORCID
Venue
Molecular Therapy - Methods & Clinical Development
Publisher
Elsevier BV
Published
2015
Volume
2
Pages
15009
ISSN
2329-0501 (ISSN-L)
DOI
10.1038/mtm.2015.9
PMC
PMC4445010
OpenAlex
W2033737576
Cited by
53
References
12
Citations by year
2026: 7 · 2025: 4 · 2024: 6 · 2023: 6 · 2022: 2 · 2021: 3 · 2020: 4 · 2019: 4 · 2018: 1 · 2017: 9 · 2016: 4 · 2015: 2 · 2014: 1
FWCI
3.67
Type
journal article
Language
en
Open access
Licence
Locations
  • Molecular Therapy — Methods & Clinical Development (publishedVersion, cc-by-nc-nd)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • Carolina Digital Repository (University of North Carolina at Chapel Hill) (submittedVersion)
  • HAL (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • UNC Libraries
Funders
Research to Prevent Blindness
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
  • Health Sciences › Medicine › Genetics › Neurogenetic and Muscular Disorders Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
Keywords
recombinant adeno-associated virus, AAV8, DYSF gene, dysferlin, muscular dystrophy, limb-girdle muscular dystrophy type 2B, disaster medical assistance team, systemic administration, intramuscular injection

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