Development of Novel Micro-dystrophins with Enhanced Functionality

Ramos, Julian N.; Hollinger, Katrin; Bengtsson, Niclas E.; Allen, J. M.; Hauschka, Stephen D.; Chamberlain, Jeffrey S.

doi:10.1016/j.ymthe.2019.01.002

Cited by 85 publications

(111 citation statements)

References 63 publications

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“…It is quite likely that these large introns had relevant impact on the evolution of the human DMD gene leading to the acquisition of new functions. Recently, new DMD microgenes designed for gene therapy and inclusive of Hinge 1, Hinge 2 and nNOS domains resulted more functional in mice (Ramos et al, 2019). Based on this knowledge, it might be important to carefully define deletion intervals by annotating intronic breakpoints to better understand how a different genomic architecture or even locus rearrangements may impact on RNA transcription, protein translation machinery, and clinical outcome.…”

Section: Dmd Deletions and Duplications: Frequency Distribution Topmentioning

confidence: 99%

The Genetic Landscape of Dystrophin Mutations in Italy: A Nationwide Study

Neri¹,

Rossi²,

Trabanelli³

et al. 2020

Front. Genet.

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in 57%, duplications in 11% and small mutations in 32%. In BMD, we found deletions in 78%, duplications in 9% and small mutations in 13%. In BMD, there are a higher number of deletions, and small mutations are more frequent than duplications. Among small mutations that are generally frequent in both phenotypes, 44% of DMD and 36% of BMD are nonsense, thus, eligible for stop codon read-through therapy; 63% of all out-of-frame deletions are eligible for single exon skipping. Patients were also assigned to Italian regions and showed interesting regional differences in mutation distribution. The full genetic characterization in this large, nationwide cohort has allowed us to draw several correlations between DMD/BMD genotype landscapes and mutation frequency, mutation types, mutation locations along the gene, exon/intron architecture, and relevant protein d o m a i n , w i t h e ff e c t s on p o p u l a t i o n ge n e t i c c h a r a c t e r i s t i c s a n d ne w personalized therapies.

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Section: Dmd Deletions and Duplications: Frequency Distribution Topmentioning

confidence: 99%

The Genetic Landscape of Dystrophin Mutations in Italy: A Nationwide Study

Neri¹,

Rossi²,

Trabanelli³

et al. 2020

Front. Genet.

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“…This patient had a distant relative who also had the deletion with a mild phenotype and studies of the localization of his truncated minigene showed the truncated protein to be correctly localized at the sarcolemma [14]. Using transgenic mdx mice as a test model of function, several groups have designed further deletions to generate microdystrophins which also allow the integration of muscle specific promoters (for reviews see [15][16][17][18]). None of the reported microgenes will function as well as the full length dystrophin as they are by necessity a compromise, although encouraging data have been reported in the dog model of the disease [19].…”

Section: Gene Therapymentioning

confidence: 99%

From diagnosis to therapy in Duchenne muscular dystrophy

Babbs

Chatzopoulou

Edwards

et al. 2020

Biochemical Society Transactions

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Genetic approaches for the diagnosis and treatment of inherited muscle diseases have advanced rapidly in recent years. Many of the advances have occurred in the treatment of Duchenne muscular dystrophy (DMD), a muscle wasting disease where affected boys are typically wheelchair bound by age 12 years and generally die in their twenties from respiratory failure or cardiomyopathy. Dystrophin is a 421 kD protein which links F-actin to the extracellular matrix via the dystrophin-associated protein complex (DAPC) at the muscle membrane. In the absence of dystrophin, the DAPC is lost, making the muscle membrane more susceptible to contraction-induced injury. The identification of the gene causing DMD in 1986 resulted in improved diagnosis of the disease and the identification of hotspots for mutation. There is currently no effective treatment. However, there are several promising genetic therapeutic approaches at the preclinical stage or in clinical trials including read-through of stop codons, exon skipping, delivery of dystrophin minigenes and the modulation of expression of the dystrophin related protein, utrophin. In spite of significant progress, the problem of targeting all muscles, including diaphragm and heart at sufficiently high levels, remains a challenge. Any therapy also needs to consider the immune response and some treatments are mutation specific and therefore limited to a subgroup of patients. This short review provides a summary of the current status of DMD therapy with a particular focus on those genetic strategies that have been taken to the clinic.

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“…This includes first the final product formulation to be delivered, with physical/physicochemical properties that may impact on each step of the delivery procedure. Considering the NA construct, its size impairs transfection efficiency, emphasizing the need for short and functional NA backbones and transgenes, such as mini‐ or micro‐dystrophin pDNA for DMD . The purity of pDNA batches is also a crucial parameter, especially regarding residual bacterial genomic DNA that can cause muscle damages .…”

Section: Other Miscellaneous Considerationsmentioning

confidence: 99%

“…For the latter application, the strong CMV promoter could be used because it confers a high expression level of the transgene in mammalian cells, whereas the duration of expression is probably not determinant . For long‐term expression applications, the use of a muscle specific promoter could be more adequate and ensure to restrict transgene expression to SM . However, it is important to note that, upon HLV, sustained expression could be obtained using various promoters (including the CMV promoter) .…”

Section: Other Miscellaneous Considerationsmentioning

confidence: 99%

“…For long‐term expression applications, the use of a muscle specific promoter could be more adequate and ensure to restrict transgene expression to SM . However, it is important to note that, upon HLV, sustained expression could be obtained using various promoters (including the CMV promoter) . The duration of transgene expression does probably also depend on the CpG dinucleotides present on pDNA because of their immunogenicity‐inducing recognition by TLR9 .…”

Section: Other Miscellaneous Considerationsmentioning

confidence: 99%

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Gene transfer to skeletal muscle using hydrodynamic limb vein injection: current applications, hurdles and possible optimizations

Guen

Gall

Midoux

et al. 2020

The Journal of Gene Medicine

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Hydrodynamic limb vein injection is an in vivo locoregional gene delivery method. It consists of administrating a large volume of solution containing nucleic acid constructs in a limb with both blood inflow and outflow temporarily blocked using a tourniquet. The fast, high pressure delivery allows the musculature of the whole limb to be reached. The skeletal muscle is a tissue of choice for a variety of gene transfer applications, including gene therapy for Duchenne muscular dystrophy or other myopathies, as well as for the production of antibodies or other proteins with broad therapeutic effects. Hydrodynamic limb vein delivery has been evaluated with success in a large range of animal models. It has also proven to be safe and welltolerated in muscular dystrophy patients, thus supporting its translation to the clinic.However, some possible limitations may occur at different steps of the delivery process. Here, we have highlighted the interests, bottlenecks and potential improvements that could further optimize non-viral gene transfer following hydrodynamic limb vein injection. K E Y W O R D Sgene transfer, hydrodynamic delivery, locoregional, non-viral gene delivery, optimizations, skeletal muscle

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Development of Novel Micro-dystrophins with Enhanced Functionality

Cited by 85 publications

References 63 publications

The Genetic Landscape of Dystrophin Mutations in Italy: A Nationwide Study

The Genetic Landscape of Dystrophin Mutations in Italy: A Nationwide Study

From diagnosis to therapy in Duchenne muscular dystrophy

Gene transfer to skeletal muscle using hydrodynamic limb vein injection: current applications, hurdles and possible optimizations

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