Systemic delivery of AAVrh74.tMCK.hCAPN3 rescues the phenotype in a mouse model for LGMD2A/R1

Sahenk, Zarife; Özeş, Burçak; Murrey, Douglas A.; Myers, Morgan; Moss, Kyle; Yalvaç, Mehmet Emir; Ridgley, Alicia; Chen, Lei; Mendell, Jerry R.

doi:10.1016/j.omtm.2021.06.010

Cited by 10 publications

(20 citation statements)

References 55 publications

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“…The pathological mechanism underlying this disease remains unclear and, currently, there is no effective therapy for these patients. To date, most therapeutic strategies for LGMDR1 are focusing on correcting the primary genetic defect through gene or cell therapies ( Roudaut et al, 2013 ; Straub and Bertoli, 2015 ; Lasa-Elgarresta et al, 2019 ; Selvaraj et al, 2019 ; Sahenk et al, 2021 ). However, there is a need for developing novel pharmacological therapies directed towards alternative targets of the disease.…”

Section: Introductionmentioning

confidence: 99%

Targeting the Ubiquitin-Proteasome System in Limb-Girdle Muscular Dystrophy With CAPN3 Mutations

Lasa-Elgarresta

Mosqueira-Martín

González-Imaz

et al. 2022

Front. Cell Dev. Biol.

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LGMDR1 is caused by mutations in the CAPN3 gene that encodes calpain 3 (CAPN3), a non-lysosomal cysteine protease necessary for proper muscle function. Our previous findings show that CAPN3 deficiency leads to reduced SERCA levels through increased protein degradation. This work investigates the potential contribution of the ubiquitin-proteasome pathway to increased SERCA degradation in LGMDR1. Consistent with our previous results, we observed that CAPN3-deficient human myotubes exhibit reduced SERCA protein levels and high cytosolic calcium concentration. Treatment with the proteasome inhibitor bortezomib (Velcade) increased SERCA2 protein levels and normalized intracellular calcium levels in CAPN3-deficient myotubes. Moreover, bortezomib was able to recover mutated CAPN3 protein in a patient carrying R289W and R546L missense mutations. We found that CAPN3 knockout mice (C3KO) presented SERCA deficits in skeletal muscle in the early stages of the disease, prior to the manifestation of muscle deficits. However, treatment with bortezomib (0.8 mg/kg every 72 h) for 3 weeks did not rescue SERCA levels. No change in muscle proteasome activity was observed in bortezomib-treated animals, suggesting that higher bortezomib doses are needed to rescue SERCA levels in this model. Overall, our results lay the foundation for exploring inhibition of the ubiquitin-proteasome as a new therapeutic target to treat LGMDR1 patients. Moreover, patients carrying missense mutations in CAPN3 and presumably other genes may benefit from proteasome inhibition by rescuing mutant protein levels. Further studies in suitable models will be necessary to demonstrate the therapeutic efficacy of proteasome inhibition for different missense mutations.

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Section: Introductionmentioning

confidence: 99%

Targeting the Ubiquitin-Proteasome System in Limb-Girdle Muscular Dystrophy With CAPN3 Mutations

Lasa-Elgarresta

Mosqueira-Martín

González-Imaz

et al. 2022

Front. Cell Dev. Biol.

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“…We examined the efficacy of NT-3 gene therapy upon muscle fiber size and fiber type composition in tibialis anterior, gastrocnemius, quadriceps, and triceps muscles at 6 months post gene injection. Succinic dehydrogenase stain was used for quantification [ 15 ], which delineates fibers according to mitochondria content: fatigue-resistant slow-twitch oxidative (STO) fibers with darkest staining, the fast-twitch oxidative (FTO) with intermediate staining and the fast-twitch glycolytic (FTG) with lightest staining intensity ( Figure 2A ). Fiber size measurements were done on the representative images from deep, intermediate, and superficial zones of untreated and NT-3-treated samples as illustrated in the tibialis anterior and gastrocnemius muscles [ 15 ] ( Figure 2B , 2C ).…”

Section: Resultsmentioning

confidence: 99%

“…Succinic dehydrogenase stain was used for quantification [ 15 ], which delineates fibers according to mitochondria content: fatigue-resistant slow-twitch oxidative (STO) fibers with darkest staining, the fast-twitch oxidative (FTO) with intermediate staining and the fast-twitch glycolytic (FTG) with lightest staining intensity ( Figure 2A ). Fiber size measurements were done on the representative images from deep, intermediate, and superficial zones of untreated and NT-3-treated samples as illustrated in the tibialis anterior and gastrocnemius muscles [ 15 ] ( Figure 2B , 2C ). Both muscles from the untreated cohort showed atrophic angular fibers with the majority belonging to fast twitch type 2 fibers, typical of sarcopenic muscle ( Figure 2A ).…”

Section: Resultsmentioning

confidence: 99%

AAV1.NT-3 gene therapy prevents age-related sarcopenia

Özeş¹,

Tong²,

Myers³

et al. 2023

Aging

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Sarcopenia is progressive loss of muscle mass and strength, occurring during normal aging with significant consequences on the quality of life for elderly. Neurotrophin 3 (NT-3) is an important autocrine factor supporting Schwann cell survival and differentiation and stimulating axon regeneration and myelination. NT-3 is involved in the maintenance of neuromuscular junction (NMJ) integrity, restoration of impaired radial growth of muscle fibers through activation of the Akt/mTOR pathway. We tested the efficacy of NT-3 gene transfer therapy in wild type (WT)-aged C57BL/6 mice, a model for natural aging and sarcopenia, via intramuscular injection 1 × 1011 vg AAV1.tMCK.NT-3, at 18 months of age. The treatment efficacy was assessed at 6 months post-injection using run to exhaustion and rotarod tests, in vivo muscle contractility assay, and histopathological studies of the peripheral nervous system, including NMJ connectivity and muscle. AAV1.NT-3 gene therapy in WT-aged C57BL/6 mice resulted in functional and in vivo muscle physiology improvements, supported by quantitative histology from muscle, peripheral nerves and NMJ. Hindlimb and forelimb muscles in the untreated cohort showed the presence of a muscle- and sex-dependent remodeling and fiber size decrease with aging, which was normalized toward values obtained from 10 months old WT mice with treatment. The molecular studies assessing the NT-3 effect on the oxidative state of distal hindlimb muscles, accompanied by western blot analyses for mTORC1 activation were in accordance with the histological findings. Considering the cost and quality of life to the individual, we believe our study has important implications for management of age-related sarcopenia.

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“…Among the parameters examined, we could see, mostly at high dose, some slight functional improvement in the young injected Gne (–/–)h GNE D207V-Tg mice for weight and grip force in the female mice group only. Indeed, better motor performance of females has been observed recently in a study on calpain 3 deficient mice [ 30 ]. This theoretically could be attributed to the different muscle fiber type distribution observed in humans between males and females [ 37 ], males presenting a more glycolytic phenotype and females a more oxidative phenotype, as also seen in transcriptome analyses [ 38 ].…”

Section: Discussionmentioning

confidence: 96%

Pre Clinical Assessment of AAVrh74.MCK.GNE Viral Vector Therapeutic Potential: Robust Activity Despite Lack of Consistent Animal Model for GNE Myopathy

Mitrani-Rosenbaum

Yakovlev

Cohen

et al. 2022

JND

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Background: GNE myopathy is a unique adult onset rare neuromuscular disease caused by recessive mutations in the GNE gene. The pathophysiological mechanism of this disorder is not well understood and to date, there is no available therapy for this debilitating disease. We have previously established proof of concept that AAV based gene therapy can effectively deliver the wild type human GNE into cultured muscle cells from human patients and in mice, using a CMV promoter driven human wild type GNE plasmid delivered through an adeno associated virus (AAV8) based platform. Objective: In the present study we have generated a muscle specific GNE construct, driven by the MCK promoter and packaged with the AAVrh74 serotype for efficacy evaluation in an animal model of GNE Myopathy. Methods: The viral vector was systemically delivered at 2 doses to two age groups of a Gne–/– hGNED207V Tg mouse described as a preclinical model of GNE Myopathy, and treatment was monitored for long term efficacy. Results: In spite of the fact that the full described characteristics of the preclinical model could not be reproduced, the systemic injection of the rAAVrh74.MCK.GNE viral vector resulted in a long term presence and expression of human wt GNE in the murine muscles and in some improvements of their mild phenotype. The Gne–/– hGNED207V Tg mice are smaller from birth, but cannot be differentiated from littermates by muscle function (grip strength and Rotarod) and their muscle histology is normal, even at advanced age. Conclusions: The rAAVrh74.MCK.GNE vector is a robust tool for the development of GNE Myopathy therapies that supply the intact GNE. However, there is still no reliable animal model to fully assess its efficacy since the previously developed Gne–/– hGNED207V Tg mice do not present disease characteristics.

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Systemic delivery of AAVrh74.tMCK.hCAPN3 rescues the phenotype in a mouse model for LGMD2A/R1

Cited by 10 publications

References 55 publications

Targeting the Ubiquitin-Proteasome System in Limb-Girdle Muscular Dystrophy With CAPN3 Mutations

Targeting the Ubiquitin-Proteasome System in Limb-Girdle Muscular Dystrophy With CAPN3 Mutations

AAV1.NT-3 gene therapy prevents age-related sarcopenia

Pre Clinical Assessment of AAVrh74.MCK.GNE Viral Vector Therapeutic Potential: Robust Activity Despite Lack of Consistent Animal Model for GNE Myopathy

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