A5084630333- Muscle Physiology and Disorders
- Virus-based gene therapy research
- RNA Interference and Gene Delivery
- CRISPR and Genetic Engineering
- Tissue Engineering and Regenerative Medicine
- RNA Research and Splicing
- Viral Infectious Diseases and Gene Expression in Insects
- Cardiomyopathy and Myosin Studies
- Adipose Tissue and Metabolism
- RNA modifications and cancer
- Nerve injury and regeneration
- CAR-T cell therapy research
- Animal Genetics and Reproduction
- Viral Infections and Immunology Research
- Mesenchymal stem cell research
- 3D Printing in Biomedical Research
- Genetic Neurodegenerative Diseases
- Herpesvirus Infections and Treatments
- Retinal Development and Disorders
- Silk-based biomaterials and applications
- RNA regulation and disease
- RNA and protein synthesis mechanisms
- Molecular Biology Techniques and Applications
- Neurogenetic and Muscular Disorders Research
- Corneal Surgery and Treatments
Inserm
2015-2024
Gene Therapy Laboratory
2011-2024
Centre Hospitalier Universitaire de Nantes
2011-2024
Translational Research in Gene Therapy
2013-2024
Nantes Université
2015-2024
Chimie et Interdisciplinarité, Synthèse, Analyse, Modélisation
2024
Institut du Thorax
2018
Genethon (France)
2010-2016
Bluebird Bio (France)
2013-2014
Institut National de Recherche en Santé Publique
2011-2013
Abstract Duchenne muscular dystrophy (DMD) is an incurable X-linked muscle-wasting disease caused by mutations in the dystrophin gene. Gene therapy using highly functional microdystrophin genes and recombinant adeno-associated virus (rAAV) vectors attractive strategy to treat DMD. Here we show that locoregional systemic delivery of a rAAV2/8 vector expressing canine (cMD1) effective restoring expression stabilizing clinical symptoms studies performed on total 12 treated golden retriever...
Recombinant adeno-associated virus (rAAV) vectors are capable of mediating long-term gene expression following administration to skeletal muscle. In rodent muscle, the vector genomes persist in nucleus concatemeric episomal forms. Here, we demonstrate with nonhuman primates that rAAV integrate inefficiently into chromosomes myocytes and reside predominantly as monomeric circles. The assimilate chromatin a typical nucleosomal pattern. persistence for years quiescent tissues suggests bona fide...
Splicing of the K-SAM alternative exon fibroblast growth factor receptor 2 gene is heavily dependent on U-rich sequence IAS1 lying immediately downstream from its 5′ splice site. We show that can activate use several heterologous sites in vitro. Addition RNA-binding protein TIA-1 to splicing extracts preferentially enhances linked IAS1. provoke a switch such pre-mRNAs with competing sites, only one which adjacent Using combination UV cross-linking and specific immunoprecipitation steps, we...
A few animal models of Duchenne muscular dystrophy (DMD) are available, large ones such as pigs or dogs being expensive and difficult to handle. Mdx (X-linked dystrophy) mice only partially mimic the human disease, with limited chronic lesions muscle weakness. Their small size also imposes limitations on analyses. rat model could represent a useful alternative since rats animals but 10 times bigger than better reflect functional abnormalities observed in DMD patients. Two lines Dmd...
Duchenne muscular dystrophy (DMD) is a severe muscle-wasting disorder caused by mutations in the dystrophin gene, without curative treatment yet available. Our study provides, for first time, overall safety profile and therapeutic dose of recombinant adeno-associated virus vector, serotype 8 (rAAV8) carrying modified U7snRNA sequence promoting exon skipping to restore functional in-frame transcript, injected locoregional transvenous perfusion forelimb. Eighteen Golden Retriever Muscular...
Decades of biological and clinical research have led to important advances in recombinant adeno-associated viruses rAAV-based gene therapy therapy. However, several challenges must be overcome fully exploit the potential rAAV vectors. Innovative approaches modify viral genome capsid elements been used issues such as unwanted immune responses off-targeting. While often successful, genetic modification capsids can drastically reduce vector yield fails produce vectors with properties that...
TIA-1 has recently been shown to activate splicing of specific pre-mRNAs transcribed from transiently transfected minigenes, and some 5' splice sites in vitro, but not any endogenous pre-mRNA. We show here that overexpression or the related protein TIAR little effect on several containing alternative exons, markedly activates normally rarely used exons pre-mRNAs. These have weak followed by U-rich stretches. When stretch following site a exon was deleted, induced use cryptic also place...
The comprehensive characterization of recombinant adeno-associated viral (rAAV) integration frequency and persistence for assessing rAAV vector biosafety in gene therapy is severely limited due to the predominance episomal genomes maintained vivo. Introducing insertional standards (rAIS), we show that linear amplification-mediated (LAM)-PCR deep sequencing can be used validated measurement frequencies. Integration rAAV2/1 or rAAV2/8, following intramuscular (IM) regional intravenous (RI)...
The robustness and safety of liver-directed gene therapy can be substantially improved by enhancing expression the therapeutic transgene in liver. To achieve this, we developed a new approach rational silico vector design. This relies on genome-wide bio-informatics strategy to identify cis-acting regulatory modules (CRMs) containing evolutionary conserved clusters transcription factor binding site motifs that determine high tissue-specific expression. Incorporation these CRMs into...
Abstract Charcot-Marie-Tooth disease 1 A (CMT1A) results from a duplication of the PMP22 gene in Schwann cells and deficit myelination peripheral nerves. Patients with CMT1A have reduced nerve conduction velocity, muscle wasting, hand foot deformations drop walking. Here, we evaluate safety efficacy recombinant adeno-associated viral vector serotype 9 (AAV2/9) expressing GFP shRNAs targeting Pmp22 mRNA animal models A. Intra-nerve delivery AAV2/9 sciatic allowed widespread transgene...
Inherited retinal diseases are a leading and untreatable cause of blindness therefore candidate for gene therapy. Recombinant vectors derived from adeno-associated virus (rAAV) currently the most promising vehicles
We developed a drug-free regional intravenous (RI) delivery protocol of recombinant adeno-associated virus (rAAV) 1 and 8 to an entire limb in the nonhuman primate (NHP), compared results with those produced by intramuscular (IM) same dose vector. show that RI both serotypes was remarkably well tolerated no adverse side-effects. After IM, muscle transduction restricted site injection high number vector copies per cell for rAAV1. In contrast, although resulted lower copy cell, it detectable...
Duchenne Muscular Dystrophy (DMD) is a fatal skeletal muscle wasting disease presenting with excessive myofibre necrosis and increased inflammation oxidative stress. In the mdx mouse model of DMD, homeostasis amino acid taurine altered, administration drastically decreases necrosis, dystropathology, protein thiol oxidation. Since severe pathology Golden Retriever (GRMD) dog more closely resembles human DMD condition, we aimed to assess generation oxidants by inflammatory cells metabolism in...
In the absence of an immune response from host, intramuscular (IM) injection recombinant adeno-associated virus (rAAV) results in permanent expression transgene mouse to primate models. However, recent gene transfer studies into animal models and humans indicate that risk and/or capsid-specific responses occurs depends on multiple factors. Among these factors, route delivery is important, although poorly addressed large Here, we compare IM drug-free regional intravenous (RI) deliveries rAAV...
Numerous studies have demonstrated the efficacy of Adeno-Associated Virus (AAV)-based gene delivery platform in vivo. The control transgene expression many protocols is highly desirable for therapeutic applications and/or safety reasons. To date, tetracycline and rapamycin dependent regulatory systems been most widely evaluated. While long-term regulation has obtained rodent models, translation these to larger animals, especially nonhuman primates (NHP), often resulted an immune response...
Anti-transgene immune responses elicited after intramuscular (i.m.) delivery of recombinant adeno-associated virus (rAAV) have been shown to hamper long-term transgene expression in large-animal models rAAV-mediated gene transfer. To overcome this hurdle, an alternative mode rAAV vectors nonhuman primate muscles has described: the locoregional (LR) intravenous route administration. Using injection mode, persistent inducible for at least 1 year under control tetracycline-inducible Tet-On...
The subretinal injection protocol for the only approved retinal gene therapy (voretigene neparvovec-rzyl) includes air tamponade at end of procedure, but its effects on bleb have not been described. In present study, we evaluated distribution enhanced green fluorescent protein (EGFP) after AAV2 in non-human primates (NHP) without (group A = 3 eyes) or with B tamponade. expression EGFP was assessed 1 month
Background: Exon skipping therapy is an emerging approach in Duchenne Muscular Dystrophy (DMD).Antisense oligonucleotides that induce of exon 51, 44, 45, or 53 are currently being evaluated clinical trials.These trials were designed on the basis data available general DMD population.Objectives: Our objective was to compare and functional statuses non-ambulant patients theoretically treatable by with other mutations.Methods: We first compared fifteen carrying deletions (DMD-53) closely...