Therapeutic Application of Extracellular Vesicles-Capsulated Adeno-Associated Virus Vector via nSMase2/Smpd3, Satellite, and Immune Cells in Duchenne Muscular Dystrophy.

Yasunari Matsuzaka,Takashi Okada,Kazuo Hashido,Yukihiko Hirai

doi:10.3390/ijms23031551

Abstract

Duchenne muscular dystrophy (DMD) is caused by loss-of-function mutations in the dystrophin gene on chromosome Xp21. Disruption of the dystrophin–glycoprotein complex (DGC) on the cell membrane causes cytosolic Ca2+ influx, resulting in protease activation, mitochondrial dysfunction, and progressive myofiber degeneration, leading to muscle wasting and fragility. In addition to the function of dystrophin in the structural integrity of myofibers, a novel function of asymmetric cell division in muscular stem cells (satellite cells) has been reported. Therefore, it has been suggested that myofiber instability may not be the only cause of dystrophic degeneration, but rather that the phenotype might be caused by multiple factors, including stem cell and myofiber functions. Furthermore, it has been focused functional regulation of satellite cells by intracellular communication of extracellular vesicles (EVs) in DMD pathology. Recently, a novel molecular mechanism of DMD pathogenesis—circulating RNA molecules—has been revealed through the study of target pathways modulated by the Neutral sphingomyelinase2/Neutral sphingomyelinase3 (nSMase2/Smpd3) protein. In addition, adeno-associated virus (AAV) has been clinically applied for DMD therapy owing to the safety and long-term expression of transduction genes. Furthermore, the EV-capsulated AAV vector (EV-AAV) has been shown to be a useful tool for the intervention of DMD, because of the high efficacy of the transgene and avoidance of neutralizing antibodies. Thus, we review application of AAV and EV-AAV vectors for DMD as novel therapeutic strategy.

Highlights

Duchenne muscular dystrophy (DMD) is a severe and progressive muscular disorder, which mainly manifests as degeneration and regeneration in skeletal and cardiac muscles and, leads to myofiber necrosis, fibrosis, and muscle weakness, through membrane fragility and disrupted cell signaling [1,2,3,4,5]
There are mainly two of treatment strategies for DMD have been explored: (1) the treatment for actual cause of DMD–dystrophin deficiency, such as adeno-associated virus (AAV)-mediated micro/minidystrophin gene delivery, synthetic antisense oligonucleotides for exon skipping, nonsense readthrough, CRISPR-Cas9-mediated genome editing, protein replacement therapies, and primarily utropin; Int
AAV vectors are considered to be a promising therapeutic strategy for treating DMD, because of their safety and usefulness, their use still has concerns, such as adverse effects caused by a single high-dose administration

Summary

Introduction

Duchenne muscular dystrophy (DMD) is a severe and progressive muscular disorder, which mainly manifests as degeneration and regeneration in skeletal and cardiac muscles and, leads to myofiber necrosis, fibrosis, and muscle weakness, through membrane fragility and disrupted cell signaling [1,2,3,4,5]. The full functional dystrophin protein forms a complex known as the dystrophinassociated protein complexes (DAPCs), with ten kinds of proteins, including the dystroglycan subcomplex (α-dystroglycan and β-dystroglycan), the sarcoglycan subcomplex (α-sarcoglycan, β-sarcoglycan, γ-sarcoglycan, and δ-sarcoglycan), sarcospan, syntrophin, dystrobrevin, and neuronal nitric oxide synthase (nNOS) [5,31,32,33,34,35,36,37,38] These differences are dependent on the type of tissues or cell and on the different regions of the same myocyte on the cell membrane. SCs lacking dystrophin markedly increase the number of abnormal nonpolarized mitotic divisions and reduced asymmetric cell divisions, exacerbating dystrophic pathology [43,44] These reports suggest that SC dysfunction is mainly involved in muscular dystrophy.

Therapy Strategy of DMD

Satellite and Immune Cells

Non-Invasive Biomarkers

Conclusions

Methods

Findings