Single Intramuscular Injection of AAV-shRNA Reduces DNM2 and Prevents Myotubular Myopathy in Mice

Tasfaout, Hichem; Lionello, Valentina M.; Kretz, Christine; Koebel, Pascale; Messaddeq, Nadia; Bitz, Deborah; Laporte, Jocelyn; Cowling, Belinda S.

doi:10.1016/j.ymthe.2018.02.008

Cited by 39 publications

(32 citation statements)

References 59 publications

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“…Adeno-associated virus (AAV) vectors have previously been used effectively to repress gene expression by encoding for shRNA complementary to the target mRNA (25). We previously screened several Dnm2 shRNA sequences that provide robust targeting of Dnm2 mRNA for degradation in vitro and in vivo (24). The best shRNA-Dnm2 sequence (AAV-sh) as well as a scrambled control sequence (AAV-C) were selected and evaluated here for their potential to knock down Dnm2 RNA in the Dnm2 RW/+ mice.…”

Section: Resultsmentioning

confidence: 99%

“…Repeated injections of ASO targeting Dnm2 into Mtm1 -/y mice efficiently reduced DNM2 and improved lifespan while reducing disease pathology (23). Most recently, using adeno-associated virus to express shRNA targeting Dnm2 in Mtm1 -/y mice, we demonstrated a robust DNM2 knockdown and disease rescue, providing an alternative strategy to reduce DNM2 (24). Therefore, targeting DNM2 was shown to be a valid potential therapy for MTM1-CNM using different approaches.…”

Section: Significancementioning

confidence: 91%

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Reducing dynamin 2 (DNM2) rescues DNM2 -related dominant centronuclear myopathy

Buono

Ross

Tasfaout

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

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Centronuclear myopathies (CNM) are a group of severe muscle diseases for which no effective therapy is currently available. We have previously shown that reduction of the large GTPase DNM2 in a mouse model of the X-linked form, due to loss of myotubularin phosphatase MTM1, prevents the development of the skeletal muscle pathophysiology. As DNM2 is mutated in autosomal dominant forms, here we tested whether DNM2 reduction can rescue DNM2-related CNM in a knock-in mouse harboring the p.R465W mutation (Dnm2RW/+) and displaying a mild CNM phenotype similar to patients with the same mutation. A single intramuscular injection of adeno-associated virus-shRNA targeting Dnm2 resulted in reduction in protein levels 5 wk post injection, with a corresponding improvement in muscle mass and fiber size distribution, as well as an improvement in histopathological CNM features. To establish a systemic treatment, weekly i.p. injections of antisense oligonucleotides targeting Dnm2 were administered to Dnm2RW/+mice for 5 wk. While muscle mass, histopathology, and muscle ultrastructure were perturbed in Dnm2RW/+mice compared with wild-type mice, these features were indistinguishable from wild-type mice after reducing DNM2. Therefore, DNM2 knockdown via two different strategies can efficiently correct the myopathy due to DNM2 mutations, and it provides a common therapeutic strategy for several forms of centronuclear myopathy. Furthermore, we provide an example of treating a dominant disease by targeting both alleles, suggesting that this strategy may be applied to other dominant diseases.

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Section: Resultsmentioning

confidence: 99%

Section: Significancementioning

confidence: 91%

Reducing dynamin 2 (DNM2) rescues DNM2 -related dominant centronuclear myopathy

Buono

Ross

Tasfaout

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

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“…First proof-of-concept results with prospective technologies such as RNA trans-splicing and RNA inhibition were recently reported. RNA trans-splicing was only tested in WT mice (20), while RNA inhibition with allele-specific shRNA silencing (21) or antisense oligonucleotides (ASO) targeting the total pool of Dnm2 both prevented the progression of the phenotypes of the Dnm2 RW/+ mouse (22,23) DNM2 mutations in ADCNM are proposed to be gain-of-function mutations. The functions of DNM2 depend on its ability to hydrolyze GTP, oligomerize, and bind lipids (24).…”

Section: Introductionmentioning

confidence: 99%

Physiological impact and disease reversion for the severe form of centronuclear myopathy linked to dynamin

Massana-Muñoz¹,

Kretz²,

Silva-Rojas³

et al. 2020

JCI Insight

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“…AAV-mediated delivery of therapeutic ASOs may represent a means to produce sustained drug delivery at a more reasonable cost. AAV delivery of shRNA and miRNA has been successfully used to decrease in vivo expression of DNM2 (myotubular myopathy) [ 78 ] and mutant huntingtin [ 79 ] in mice. SOD1 was successfully targeted for reduction with an AAV-delivered exon skipping ASO in a murine model of amyotrophic lateral sclerosis [ 80 ].…”

Section: Gene Therapy: Brain Tropic Adenoviral Vectorsmentioning

confidence: 99%

Emerging Technologies for Delivery of Biotherapeutics and Gene Therapy Across the Blood–Brain Barrier

2018

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Antibody, immuno- and gene therapies developed for neurological indications face a delivery challenge posed by various anatomical and physiological barriers within the central nervous system (CNS); most notably, the blood–brain barrier (BBB). Emerging delivery technologies for biotherapeutics have focused on trans-cellular pathways across the BBB utilizing receptor-mediated transcytosis (RMT). ‘Traditionally’ targeted RMT receptors, transferrin receptor (TfR) and insulin receptor (IR), are ubiquitously expressed and pose numerous translational challenges during development, including species differences and safety risks. Recent advances in antibody engineering technologies and discoveries of RMT targets and BBB-crossing antibodies that are more BBB-selective have combined to create a new preclinical pipeline of BBB-crossing biotherapeutics with improved efficacy and safety. Novel BBB-selective RMT targets and carrier antibodies have exposed additional opportunities for re-targeting gene delivery vectors or nanocarriers for more efficient brain delivery. Emergence and refinement of core technologies of genetic engineering and editing as well as biomanufacturing of viral vectors and cell-derived products have de-risked the path to the development of systemic gene therapy approaches for the CNS. In particular, brain-tropic viral vectors and extracellular vesicles have recently expanded the repertoire of brain delivery strategies for biotherapeutics. Whereas protein biotherapeutics and bispecific antibodies enabled for BBB transcytosis are rapidly heading towards clinical trials, systemic gene therapy approaches for CNS will likely remain in research phase for the foreseeable future. The promise and limitations of these emerging cross-BBB delivery technologies are further discussed in this article.

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Single Intramuscular Injection of AAV-shRNA Reduces DNM2 and Prevents Myotubular Myopathy in Mice

Cited by 39 publications

References 59 publications

Reducing dynamin 2 (DNM2) rescues DNM2 -related dominant centronuclear myopathy

Reducing dynamin 2 (DNM2) rescues DNM2 -related dominant centronuclear myopathy

Physiological impact and disease reversion for the severe form of centronuclear myopathy linked to dynamin

Emerging Technologies for Delivery of Biotherapeutics and Gene Therapy Across the Blood–Brain Barrier

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