Dystrophins, Utrophins, and Associated Scaffolding Complexes: Role in Mammalian Brain and Implications for Therapeutic Strategies

Two decades of molecular, cellular, and functional studies considerably increased our understanding of dystrophins function and unveiled the complex etiology of the cognitive deficits in Duchenne muscular dystrophy (DMD), which involves altered expression of several dystrophin-gene products in brain. Dystrophins are normally part of critical cytoskeleton-associated membrane-bound molecular scaffolds involved in the clustering of receptors, ion channels, and signaling proteins that contribute to synapse physiology and blood-brain barrier function. The utrophin gene also drives brain expression of several paralogs proteins, which cellular expression and biological roles remain to be elucidated. Here we review the structural and functional properties of dystrophins and utrophins in brain, the consequences of dystrophins loss-of-function as revealed by numerous studies in mouse models of DMD, and we discuss future challenges and putative therapeutic strategies that may compensate for the cognitive impairment in DMD based on experimental manipulation of dystrophins and/or utrophins brain expression

Investigating the Impact of Delivery Routes for Exon Skipping Therapies in the CNS of DMD Mouse Models

Nucleic acid-based therapies have demonstrated great potential for the treatment of monogenetic diseases, including neurologic disorders. To date, regulatory approval has been received for a dozen antisense oligonucleotides (ASOs); however, these chemistries cannot readily cross the blood–brain barrier when administered systemically. Therefore, an investigation of their potential effects within the central nervous system (CNS) requires local delivery. Here, we studied the brain distribution and exon-skipping efficacy of two ASO chemistries, PMO and tcDNA, when delivered to the cerebrospinal fluid (CSF) of mice carrying a deletion in exon 52 of the dystrophin gene, a model of Duchenne muscular dystrophy (DMD). Following intracerebroventricular (ICV) delivery (unilateral, bilateral, bolus vs. slow rate, repeated via cannula or very slow via osmotic pumps), ASO levels were quantified across brain regions and exon 51 skipping was evaluated, revealing that tcDNA treatment invariably generates comparable or more skipping relative to that with PMO, even when the PMO was administered at higher doses. We also performed intra-cisterna magna (ICM) delivery as an alternative route for CSF delivery and found a biased distribution of the ASOs towards posterior brain regions, including the cerebellum, hindbrain, and the cervical part of the spinal cord. Finally, we combined both ICV and ICM injection methods to assess the potential of an additive effect of this methodology in inducing efficient exon skipping across different brain regions. Our results provide useful insights into the local delivery and associated efficacy of ASOs in the CNS in mouse models of DMD. These findings pave the way for further ASO-based therapy application to the CNS for neurological disease

Genes, Plasticity and Mental Retardation.

International audienceFunctional and structural plasticity is a fundamental property of the brain involved in diverse processes ranging from brain construction and repair to storage of experiences during lifetime. Our current understanding of different forms of brain plasticity mechanisms has advanced tremendously in the last decades, benefiting from studies of development and memory storage in adulthood and from investigations of diverse diseased conditions. In this review, we focus on the role of mental retardation (MR) genes and show how this developing area of research can enrich our knowledge of the cellular and molecular mechanisms of brain plasticity and cognitive functions, and of the dysfunctional mechanisms underlying MR. We describe two main groups of MR genes; those leading to dysfunctional neurodevelopmental programs and brain malformations, and those which rely on alterations in molecular mechanisms underlying synaptic organization and plasticity. We first explore the role of MR genes in key mechanisms of neurogenesis and neuronal migration during development and in the adult, such as actin and microtubule-cytoskeletal dynamics and signal transduction. We then define the contribution of MR genes to forms of activity-dependent synaptic modifications, such as those involved in molecular organization of the synapse, intracellular signaling regulating gene programs and neuronal cytoskeleton to control network remodeling. We trace the characteristics of MR genes playing key roles in many forms of brain plasticity mechanisms, and highlight specific MR genes that endorse distinct roles in different cell types or brain regions, and at various times of a brain lifetime

Gene Therapy for Central Nervous System in Duchenne Muscular Dystrophy

International audienceThe development of molecular therapies enabling compensation of brain alterations in Duchenne muscular dystrophy is a major objective given the high level of functional impairment associated with intellectual disability and neuropsychiatric disorders in this syndrome. Functional and preclinical studies in mice lacking distinct brain dystrophins identified an accurate set of phenotypes, from the molecular to neurophysiological and behavioral levels, which can be used as markers of efficacy for brain gene therapy. Pioneer studies in this past decade provided encouraging results, demonstrating that both dmd-gene splice-switching correction and replacement strategies hold realistic prospects to rescue expression and function of the brain full-length (Dp427) or short C-terminal (Dp71) dystrophins responsible for variable degrees of cognitive impairment in DMD. Strategies that could correct or alleviate both muscle and brain dysfunctions entail selection of molecular tools able to cross the blood-brain barrier following systemic delivery, to largely spread in neural tissues, and to selectively target the neural cell types (neurons, astrocytes) that require rescued expression of distinct brain dystrophins. Recent breakthroughs show that this can be achieved by engineering naked antisense oligonucleotides with specific chemistries and/or adeno-associated virus vectors with selective capsid properties, thus raising new hopes to bring gene therapy closer to whole-body delivery and full treatment of DMD symptoms

Facilitated NMDA receptor-mediated synaptic plasticity in the hippocampal CA1 area of dystrophin-deficient mice

International audienceThe contribution of the cytoskeletal membrane-associated protein dystrophin in glutamatergic transmission and related plasticity was investigated in the hippocampal CA1 area of wild-type and dystrophin-deficient (mdx) mice, using extracellular recordings in the ex vivo slice preparation. Presynaptic fiber volleys and field excitatory postsynaptic potentials (fEPSPs) mediated through N-methyl-D-Aspartate receptors (NMDAr) or non-NMDAr were compared in both strains. Comparable synaptic responses were observed in wild-type and mdx mice, suggesting that basal glutamatergic transmission is not altered in the mutants. By contrast, the synaptic strengthening induced by a conditioning stimulation of either 10, 30, or 100 Hz was significantly greater in mdx mice during the first minutes posttetanus. Because the posttetanic potentiation induced in the presence of the NMDAr antagonist D-APV was not affected in the mutants, a critical role of NMDAr in this increase was suggested. The magnitude of the potentiation induced by a 30 Hz stimulation in mdx mice was normalized as compared to wild-type mice by increasing the extracellular magnesium concentration from 1.5 to 3 mM. Moreover, the transitory depression of fEPSPs induced by bath-applied NMDA (50 microM for 30s) was more sensitive to an increased extracellular magnesium concentration in wild-type than in mdx mice. Our results suggest that the absence of dystrophin may facilitate NMDAr activation in the CA1 hippocampal subfield of mdx mice, which may be partly due to a reduction of the voltage-dependent block of this receptor by magnesium

Impaired long-term spatial and recognition memory and enhanced CA1 hippocampal LTP in the dystrophin-deficient Dmdmdx mouse

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Facilitated NMDA receptor-mediated synaptic plasticity in the hippocampal CA1 area of dystrophin-deficient mice

International audienc