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ANR funded project

Une nouvelle représentation du vivant (DS0401)
Edition 2014


Dystrophins In The Nervous System: From Neurophysiology to Molecular Therapy

Molecular tools for gene therapy in the central nervous system and for muscular dystrophy
Central alterations in muscular dystrophy involve distinct products of the dystrophin gene expressed in neurons and/or in glial cells. The development of specific molecular tools is required to study physiopathological mechanisms and to develop treatments targetting the central nervous system.

Unravelling and correcting neuronal and glial alterations in muscular dystrophy
Biotechnologies have open new routes for the treatment of Duchenne muscular dystrophy, but only few studies evaluated the potential of these treatments to correct central dysfunctions in this disease, yet they may lead to severe intellectual and neuropsychiatric disturbances. To target central defects in this syndrome, two main challenges need to be taken: (1) To better understand the mechanisms responsible for clinical heterogeneity, which depends on the variety of mutations leading to alteration of several forms of dystrophins implicated in distinct neuronal and glial functions and (2) to design molecular tools adapted to target specific cell types and/or to treat more largely the different affected organs.
To acheive these goals, we propose a multidisciplinary approach, from gene to behavior, of the physiopathological mechanisms displayed by mouse genetic models baring a specific loss of the neuronal or glial dystrophin. The therapeutical approaches are based on the administration of antisens oligonucleotides allowing elimination of the mutation or of transgenes providing ectopic expression of the lost protein, through conveyance by specific vectors designed to target neurons and glia, or through coupling with specific chemistries that favor crossing into nervous tissues following systemic administration. Identification of new neuropathological mechanisms and efficient molecular tools for gene therapy will favor breakthroughs into developable therapeutic approaches and novel treatment opportunities for a range of genetic diseases featuring alterations of the central nervous system.

Identification of neuropathological markers and development of tools for therapy
The DYSther project combines expertise of four teams, from molecular to behavior, forming a multidisciplinary network to tackle emerging questions that link fundamental research to technological challenges aimed at improving research tools and translational rescue strategies. Our experimental approach, undertaken in mouse models of the human disease, is based on converging and complementary methods including cellular and molecular biology, biochemistry, neuroanatomy, brain and cell imaging, pharmacology, electrophysiology and behavioral exploration. We also use models in which specific genes facilitate cell imaging or enable activation of specific neuronal networks by optogenetic technology, or modulate inflammation of nervous tissues to study underlying risk factors in this process.
The numerous data generated by this approach enable identification of specific biomarkers to be used for relevant evaluation of the efficacy of the molecular tools that we develop and produce in large amounts for therapeutical strategies. To help these molecules to cross barriers and diffuse in all affected regions of the central nervous system, we pair them with particular chemistries or we include them in specific vectors selected for their potential to target specific cell types.


Identification of the behavioral defects due to the loss of the neuronal (Dp427) or mainly glial (Dp71) forms of dystrophins is achieved and partly published. The Dp427-deficient mdx mouse displays enhanced stress reactivity, altered social behavior and deficient long-term memory. The molecular tools developed to treat muscular dystrophy through Dp427 rescue have been optimized so that they can also produce beneficial effects in brain. Enhanced stress reactivity constitutes a relevant marker for rapid evaluation of treatment effects on mdx mice behavior. Dp71, a short form of dystrophin associated with severe intellectual disturbances in certain patients, is mainly expressed in glial cells and plays a role in the mechanisms that maintain brain ion equilibrium and regulate the capacity of circulating molecules to penetrate nervous tissues. Our recent studies uncovered major alterations in the number and morphology of glial cells and in the organization of the vascular network in the retina. The molecular, physiological and behavioral modifications due to the loss of Dp71 in the brain are currently being investigated. We also have selected a gene-transfer vector for future studies aimed at rescuing Dp71 expression in glial cells to compensate these alterations. The dysfunctions identified in these models will serve as markers to evaluate efficacy of the molecular tools developed for therapeutic approaches. Optimal doses of therapeutic molecules and vectors required for this work have been charaterized in this first part of the project.


The DYSther project will bring new knowledge on central nervous system functioning, on the mechanisms shared by retina and brain and on the determination of novel markers to test treatment efficacy on central dysfunctions in muscular dystrophy. Our main goal is to identify and produce molecular tools bringing gene therapy closer to a full compensation of symptoms in muscular dystrophy patients. We also aim at identifying the most reliable molecular tools and optimal conditions to correct defective genes or to compensate their dysfunction by gene therapy. This reasearch should therefore have a wide impact on numerous genetic diseases involving central nervous system dysfunctions leading to sensory, cognitive and neuropsychiatric disturbances.

Scientific outputs and patents

We found that absence of the full-length neuronal form of dystrophin (Dp427) in mdx mice leads to behavioral modifications that may underlie part of cognitive and neuropsychiatric disturbances in muscular dystrophy (Miranda et al., Mol Autism, 2015). The molecular tools developed for therapeutic intervention in this model have been optimized (Relizani et al., in preparation). In line with the expected role of Dp71 in glia, Dp71 loss induce major changes in retinal astrocytes (Giocanti-Aurégan et al., Glia, 2015). This work also led to several communications in international congresses or within science popularization interventions.



IDV Institut de la Vision

PSUD/IBBMC Université paris-Sud/Institut de Biochimie et Biophysique Moléculaire et Cellulaire


ANR grant: 540 532 euros
Beginning and duration: novembre 2014 - 36 mois

Submission abstract

Recent advances in our understanding of dystrophin-dependent physiological mechanisms responsible for Duchenne muscular dystrophy (DMD) led to the development of innovative tools for molecular therapies. Much less is known, however, on their potency to alleviate the brain and cognitive impairments associated with DMD. We previously showed that gene-therapy strategies, such as antisense-mediated exon skipping, hold realistic prospects of restoring both muscle and brain functions in the dystrophin(Dp427)-deficient mdx mouse. However, their efficacy to correct behavioral deficits was not estimated, as viral vectors precluded body-wide gene delivery. Our progresses in developing new chemistries of oligonucleotide analogs and new generations of adeno-associated vectors should overcome such limitations, as attested by our preliminary results in mdx mice. However, the intellectual disabilities in DMD are associated with mutations that lead to a cumulative loss of several dystrophin products encoded by distinct internal promoters, such as the Dp71 protein, which loss is a pivotal aggravating factor for cognitive status in DMD. A major challenge to achieve a full compensation of motor and cognitive dysfunctions in DMD therefore involves identification of reliable biomarkers and target mechanisms affected by the loss of both Dp427 and Dp71 and the development of appropriate molecular tools to rescue their respective functions.
The DYSther project combines expertise of four teams, from the molecular to the physiological and behavioral levels, forming a multidisciplinary network to tackle emerging questions that link fundamental research to technological challenges aimed at improving research tools and translational rescue strategies. DYSther is aimed at better understanding the multiple roles of dystrophin products in the CNS and to validate biomarkers for molecular therapies. Task 1 will characterize the deficits in high-order cognitive functions in mice lacking Dp427 or Dp71, with behavioral tests refined from a translational dialogue with clinicians. This will constitute the behavioral read-outs of the efficacy of gene therapy tools, including exon-skipping using AAV-U7 vectors and/or new generations of antisense oligonucleotide analogs that cross over the blood-brain barrier, and cell-specific re-expression of Dp71. These vectors will also serve to understand the implication of various brain structures and cellular subtypes in establishing the phenotype, through local delivery and use of cell-specific promoters. Task 2 will bring together concepts from brain and retina studies to decipher the role of Dp71 in glial-vascular interactions in normal and challenged conditions. Task3 will determine the impact of Dp71 deficiency on glial physiology in cerebellum, a major structure involved in the cognitive impairment in DMD. Functional similarities between retinal Müller and cerebellar Bergmann glial cells suggest that most hypotheses derived from retina studies could be tested in cerebellar tissues. We will particularly focus on the Dp71-dependent mechanisms whereby astrocytes control extracellular K+ concentration and water balance in both normal and ischemic conditions. This will be completed by in vivo magnetic resonance imaging (MRI) studies of the dynamics and recovery of water accumulation during ischemia/hypoxia and brain edema. These two tasks will also generate many new read-outs that we will use to assess therapeutic rescue and modulation of Dp71 expression in both normal and pathological conditions.
The DYSther project will thus bring fundamental knowledge on the roles of dystrophins in CNS, while bringing gene therapy closer to a full compensation of symptoms in Duchenne patients. Expected breakthroughs in the mechanisms underlying glial-vascular functions, as well as in the technological capacity of new molecular tools to cross the nervous system barriers, is also of critical interest for several pathologies other than DMD.


ANR Programme: Une nouvelle représentation du vivant (DS0401) 2014

Project ID: ANR-14-CE13-0037

Project coordinator:
Monsieur Cyrille Vaillend (CNRS UMR8195)


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The project coordinator is the author of this abstract and is therefore responsible for the content of the summary. The ANR disclaims all responsibility in connection with its content.