Systemic delivery of an AAV9 exon-skipping vector significantly improves or prevents features of Duchenne muscular dystrophy in the Dup2 mouse

Wein, Nicolas; Vetter, Tatyana A.; Vulin, Adeline; Simmons, Tabatha R.; Frair, E.; Bradley, A.; Gushchina, Liubov V.; Almeida, Camila de; Huang, Nianyuan; Lesman, Daniel; Rajakumar, Dhanarajan; Weiss, Robert B.; Flanigan, Kevin M.

doi:10.1016/j.omtm.2022.07.005

Cited by 22 publications

(20 citation statements)

References 43 publications

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“…AAV delivered U7-snRNAs are currently in clinical trials for DMD as an exon skipping therapy for exon 2 duplications, and have been used pre-clinically in fascioscapulohumeral muscular dystrophy (FSHD) to reduce DUX4 expression by similarly targeting the PAS. 17,18 Aside from showing reduction of DNAJB6 isoform levels at the RNA and protein levels in primary mouse myotube cultures and primary LGMDD1 patient derived myoblasts, we demonstrated selective knockdown of DNAJB6 isoform protein levels in the TA muscle of mice with IM injections. We also provided preliminary evidence of safety against potential off-target effects.…”

Section: Discussionmentioning

confidence: 81%

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DNAJB6 Isoform Specific Knockdown: Therapeutic Potential for LGMDD1

Findlay

Paing

Daw

et al. 2022

Preprint

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Dominant missense mutations in DNAJB6, an HSP40 co-chaperone, cause limb girdle muscular dystrophy (LGMD) D1. No treatments are currently available. Two isoforms exist, DNAJB6a and DNAJB6b, each with distinct localizations in muscle. Mutations reside in both isoforms, yet evidence suggests only DNAJB6b is responsible for disease pathogenesis. Mechanistic data supports either a toxic gain of function, a dominant negative mechanism, or a combination of both. Knockdown treatment strategies involving both isoforms carry risk as DNAJB6 knockout is embryonic lethal. We therefore developed an isoform specific knockdown approach using morpholinos. Selective reduction of each isoform was achieved in-vitro in primary mouse myotubes and human myoblasts, as well as in-vivo in mouse skeletal muscle. To assess isoform specific knockdown in LGMDD1, we created primary myotube cultures from a knock-in LGMDD1 mouse model. Using mass spectrometry, we identified an LGMDD1 protein signature related to protein homeostasis and myofibril structure. Selective reduction of DNAJB6b levels in LGMDD1 myotubes corrected much of the proteomic disease signature towards wild type levels. While additional in-vivo functional data is required, these findings suggest selective reduction of DNAJB6b may be a viable therapeutic target for LGMDD1.

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Section: Discussionmentioning

confidence: 81%

Section: Discussionmentioning

confidence: 99%

DNAJB6 Isoform Specific Knockdown: Therapeutic Potential for LGMDD1

Findlay

Paing

Daw

et al. 2022

Preprint

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“…This therapy strays away from direct AON treatment, instead opting for treatment with an adeno-associated virus (AAV) vector which allows for the endogenous production of non-coding U7 small nuclear RNAs (U7snRNAs) [34]. These U7snRNAs act in a similar fashion to AONs, inducing exon 2 skipping by targeting splice donor and acceptor sites on the pre-mRNA flanking exon 2 [35]. This leads to the production of normal, full-length dystrophin transcripts and proteins.…”

Section: Atl1102mentioning

confidence: 99%

“…Pre-clinical studies in mice identified that SCAAV9.U7.ACCA was able to effectively skip one of the duplicate copies of exon 2, restoring the reading frame and enabling dystrophin production [35,36]. The researchers also found that both copies of exon 2 are occasionally skipped, resulting in an out-of-frame dystrophin transcript [37].…”

Section: Atl1102mentioning

confidence: 99%

Recent Trends in Antisense Therapies for Duchenne muscular dystrophy

Wilton-Clark¹,

Yokota²

2023

Preprint

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Duchenne muscular dystrophy (DMD) is a debilitating and fatal genetic disease affecting 1/3500 boys globally, characterized by progressive muscle breakdown and eventual death with an average lifespan in the mid-late twenties. While no cure yet exists for DMD, gene and antisense therapies have been heavily explored in recent years to better treat this disease. Four antisense therapies have received conditional FDA approval, and many more exist in varying stages of clinical trials. These upcoming therapies often utilize novel drug chemistries to address limitations of existing therapies, and their development could herald the next generation of antisense therapy. This review article aims to summarize the current state of development for antisense-based therapies for the treatment of Duchenne muscular dystrophy, exploring candidates designed for both exon skipping and gene knockdown.

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“…U7 snRNAs have the advantage of being protected from degradation by forming small nuclear ribonucleoprotein (snRNP) complexes and accumulating in the nucleus while being non-toxic and non-immunologic for cells. Encapsidation of vectors containing U7 snRNA sequences promoting exon skipping in AAVs and their systemic delivery in mouse and dog models of DMD have been reported to alleviate the dystrophic phenotype in multiple studies [ 182 , 183 , 184 , 185 , 186 ]. Importantly, the stable expression of the U7 snRNA allowed partial restoration of full-length dystrophin expression for more than 1 year after the injection [ 183 , 184 ].…”

Section: Alternative Rna-based Strategies For Treating Dm1mentioning

confidence: 99%

Recent Progress and Challenges in the Development of Antisense Therapies for Myotonic Dystrophy Type 1

Serres-Bérard

Benichou

Jauvin

et al. 2022

IJMS

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Myotonic dystrophy type 1 (DM1) is a dominant genetic disease in which the expansion of long CTG trinucleotides in the 3′ UTR of the myotonic dystrophy protein kinase (DMPK) gene results in toxic RNA gain-of-function and gene mis-splicing affecting mainly the muscles, the heart, and the brain. The CUG-expanded transcripts are a suitable target for the development of antisense oligonucleotide (ASO) therapies. Various chemical modifications of the sugar-phosphate backbone have been reported to significantly enhance the affinity of ASOs for RNA and their resistance to nucleases, making it possible to reverse DM1-like symptoms following systemic administration in different transgenic mouse models. However, specific tissue delivery remains to be improved to achieve significant clinical outcomes in humans. Several strategies, including ASO conjugation to cell-penetrating peptides, fatty acids, or monoclonal antibodies, have recently been shown to improve potency in muscle and cardiac tissues in mice. Moreover, intrathecal administration of ASOs may be an advantageous complementary administration route to bypass the blood-brain barrier and correct defects of the central nervous system in DM1. This review describes the evolution of the chemical design of antisense oligonucleotides targeting CUG-expanded mRNAs and how recent advances in the field may be game-changing by forwarding laboratory findings into clinical research and treatments for DM1 and other microsatellite diseases.

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Systemic delivery of an AAV9 exon-skipping vector significantly improves or prevents features of Duchenne muscular dystrophy in the Dup2 mouse

Cited by 22 publications

References 43 publications

DNAJB6 Isoform Specific Knockdown: Therapeutic Potential for LGMDD1

DNAJB6 Isoform Specific Knockdown: Therapeutic Potential for LGMDD1

Recent Trends in Antisense Therapies for Duchenne muscular dystrophy

Recent Progress and Challenges in the Development of Antisense Therapies for Myotonic Dystrophy Type 1

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