Academic Journal
Assessment of Therapeutic Potential of a Dual AAV Approach for Duchenne Muscular Dystrophy
| العنوان: | Assessment of Therapeutic Potential of a Dual AAV Approach for Duchenne Muscular Dystrophy |
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| المؤلفون: | Albini, Sonia, Palmieri, Laura, Dubois, Auriane, Bourg, Nathalie, Lostal, William, Richard, Isabelle |
| المساهمون: | Approches génétiques intégrées et nouvelles thérapies pour les maladies rares (INTEGRARE), Université d'Évry-Val-d'Essonne (UEVE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay-Généthon |
| المصدر: | ISSN: 1661-6596. |
| بيانات النشر: | HAL CCSD MDPI |
| سنة النشر: | 2023 |
| المجموعة: | Archive ouverte HAL (Hyper Article en Ligne, CCSD - Centre pour la Communication Scientifique Directe) |
| مصطلحات موضوعية: | gene therapy AAV dual vector DMD dystrophin homologous recombination concatemerization, gene therapy, AAV, dual vector, DMD, dystrophin, homologous recombination, concatemerization, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology |
| الوصف: | Duchenne muscular dystrophy (DMD) is a yet incurable rare genetic disease that affects the skeletal and cardiac muscles, leading to progressive muscle wasting and premature death. DMD is caused by the lack of dystrophin, a muscle protein essential for the biochemical support and integrity of muscle fibers. Gene replacement strategies for Duchenne muscular dystrophy (DMD) employing the adeno-associated virus (AAV) face the challenge imposed by the limited packaging capacity of AAV, only allowing the accommodation of a short version of dystrophin (µDys) that is still far removed from correcting human disease. The need to develop strategies leading to the expression of a best performing dystrophin variant led to only few studies reporting on the use of dual vectors, but none reported on a method to assess in vivo transgene reconstitution efficiency, the degree of which directly affects the use of safe AAV dosing. We report here on the generation of a dual AAV vector approach for the expression of a larger dystrophin version (quasidystrophin) based on homologous recombination, and the development of a methodology employing a strategic droplet digital PCR design, to determine the recombination efficiency as well as the occurrence of unwanted concatemerization events or aberrant expression from the single vectors. We demonstrated that, upon systemic delivery in the dystrophic D2.B10-Dmd mdx /J (DBA2mdx) mice, our dual AAV approach led to high transgene reconstitution efficiency and negligible Inverted Terminal Repeats (ITR)-dependent concatemerization, with consequent remarkable protein restoration in muscles and improvement of muscle pathology. This evidence supports the suitability of our system for gene therapy application and the potential of this methodology to assess and improve the feasibility for therapeutic translation of multiple vector approaches. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| Relation: | hal-04302872; https://hal.science/hal-04302872; https://hal.science/hal-04302872/document; https://hal.science/hal-04302872/file/ijms-24-11421-v2.pdf |
| DOI: | 10.3390/ijms241411421 |
| الاتاحة: | https://hal.science/hal-04302872 https://hal.science/hal-04302872/document https://hal.science/hal-04302872/file/ijms-24-11421-v2.pdf https://doi.org/10.3390/ijms241411421 |
| Rights: | info:eu-repo/semantics/OpenAccess |
| رقم الانضمام: | edsbas.1DA45323 |
| قاعدة البيانات: | BASE |
| DOI: | 10.3390/ijms241411421 |
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