Assessment of physiological parameters in the application of a double adeno-associated virus 9 with a codon-optimized DYSF gene for limb girdle muscular dystrophy type R2

Authors

DOI:

https://doi.org/10.18413/rrpharmacology.11.642

Abstract

Introduction: Gene therapy for Myoshi myopathy is extremely relevant, as it may become the first pathogenetic treatment for dysferlinopathy. The aim of this study was to study the efficacy and safety of the use of a genetic construct, the AAV9-DYSF-DV3’ virus, for the treatment of limb girdle muscular dystrophy LGMD) type R2.

Materials and Methods: Mouse models of limb girdle muscular dystrophy type R2 В6.А-Dysfprmd/GeneJ were used to study the effectiveness of AAV9.DYSF drug and the corresponding C57BL/6J controls were used. During the study, muscle activity was determined on the basis of the following tests: “Grip test”, “Holding an animal on a slippery surface of a vertical rod”, “Forced swimming with a load”, and ”Wire hanging”. In the course of acute and subchronic toxicity, hematological and biochemical blood tests of the rats, histological analysis and ”Open field” behavioral testing were performed.

Results and Discussion: In this study, for the first time, a comprehensive investigation of the effectiveness of gene therapy using the two-vector system of adeno-associated AAV9-DYSF-DV3’ virus with overlapping DYSF cDNA sequences was conducted in a mouse model of limb girdle muscular dystrophy type 2 R.

Conclusion: During the testing of the drug’s effectiveness, it was discovered that drug AAV9.DYSF showed the best effectiveness in mice with the absence of the protein dysferlin in behavioral testing at the maximum dose (5*1012) with a double intramuscular injection. In the “Grip test”, the index in В6.А-Dysfprmd/GeneJ mice increased by 29% (p=0.0026) relative to that in the K-group. In the tests “Forced swimming with a load”, ”Wire hanging”, and ”Holding an animal on a slippery surface of a vertical rod”, the indicators also improved by 80% (p=0.0019), 104.8% (p=0.001) and 20% (p=0.025), respectively, relative to those of the negative control. During acute and subchronic toxicity, the administration of the drug to animals does not cause death or intoxication.

Graphical Abstract

Keywords:

dysferlin, LGMD R2, dual AAV vector, gene transfer, muscle regeneration

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Author Contribution

Elеna V. Kuzubova, Belgorod State National Research University

Junior Researcher at the Laboratory of Genetic Technologies and Gene Editing for Biomedicine and Veterinary Science, Belgorod State National Research University, Belgorod, Russia; e-mail: 1015artek1015@mail.ru; ORCID ID: https://orcid.org/0000-0003-2425-5027. Writing an article, processing the results obtained, and conducting behavioral tests.

Alexandra I. Radchenko, Belgorod State National Research University

Junior Researcher at the Laboratory of Genetic Technologies and Gene Editing for Biomedicine and Veterinary Science, Belgorod State National Research University, Belgorod, Russia; e-mail: sandrinkaradchenko@gmail.com;ORCID ID: https://orcid.org/0000-0002-4554-2116. Conducting behavioral tests.

Andrey А. Manuylov, Belgorod Federal Agrarian Scientific Center of the Russian Academy of Sciences

Cand. Sci. (Geography), Junior Researcher at the Analytical Laboratory, Belgorod Federal Agrarian Scientific Center of the Russian Academy of Sciences, Belgorod, Russia; e-mail: infa152@yandex.ru; ORCID ID: https://orcid.org/0009-0008-7306-1919. Conducting animal genotyping and PCR.

Ariana M. Korokina, Belgorod State National Research University

Student of the Medical institute, Belgorod State National Research University, Belgorod, Russia; e-mail: ariana.korokina@mail.ru; ORCID ID: https://orcid.org/0009-0009-3265-5697. Preparating animals cohorts.

Natalia V. Koroleva, Belgorod State National Research University

Research Associate of the Department of Pharmacology and Clinical Pharmacology, Belgorod State National Research University, Belgorod, Russia; e-mail: nkoroleva@gmail.com; ORCID ID: https://orcid.org/0009-0006-3660-3438. Editing the text of the article.

Ivan A. Yakovlev, Genotaget LLC

CEO at Genotarget LLP, Moscow, Russia; е-mail: ivan@ivan-ya.ru; ORCID ID: https://orcid.org/0000-0001-8127-4078. Design development and research program writing.

Arthur A. Isaev, Artgen Biotech

Chairman of the Board of Directors at Artgen Biotech, Moscow, Russia; e-mail: art.isaev@gmail.com; ORCID ID: https://orcid.org/0000-0001-5848-5117. Literature analysis, validating the experimental methodology and participating in the planning of the experiment.

Roman V. Deev, Avtsyn Research Institute of Human Morphology of Petrovsky National Research Centre of Surgery

Candidate of Sciences (Medicine), Deputy Director at Avtsyn Research Institute of Human Morphology, Petrovsky National Research Centre of Surgery, Moscow, Russia; e-mail: romdey@gmail.com; ORCID ID: https://orcid.org/0000-0001-8389-3841. Consultation on planning, methodology and experiment implementation.

Mikhail V. Korokin, Belgorod State National Research University

Doct. Sci. (Medicine), Professor at the Department of Pharmacology and Clinical Pharmacology, Belgorod State National Research University, Belgorod, Russia; e-mail: mkorokin@mail.ru; ORCID ID: https://orcid.org/0000-0001-5402-0697. Validating the experimental methodology and taking part in the planning of the experiment.

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Published

23-06-2025

How to Cite

Kuzubova EV, Radchenko AI, Manuylov AА, Korokina AM, Koroleva NV, Yakovlev IA, Isaev AA, Deev RV, Korokin MV (2025) Assessment of physiological parameters in the application of a double adeno-associated virus 9 with a codon-optimized DYSF gene for limb girdle muscular dystrophy type R2. Research Results in Pharmacology 11(2): 36–47. https://doi.org/10.18413/rrpharmacology.11.642

Issue

Vol. 11 No. 2 (2025)

Section

Experimental Pharmacology

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Copyright (c) 2025 Kuzubova EV, Radchenko AI, Manuylov AA, Korokina AM, Koroleva NV, Yakovlev IA, Isaev AA, Deev RV, Korokin MV

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This work is licensed under a Creative Commons Attribution 4.0 International License.

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