Dystrophin R16/17 protein therapy restores sarcolemmal nNOS in trans and improves muscle perfusion and function

Molecular Medicine

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[1] 0000 0001 2162 3504, grid.134936.a, Department of Biomedical Sciences, College of Veterinary Medicine, University of Missouri, 65212, Columbia, MO, USA;0000 0001 2162 3504, grid.134936.a, Department of Molecular Microbiology and Immunology, School of Medicine, University of Missouri, Medical Sciences Building, One Hospital Drive, 65212, Columbia, MO, USA;0000 0001 2162 3504, grid.134936.a, Department of Molecular Microbiology and Immunology, School of Medicine, University of Missouri, Medical Sciences Building, One Hospital Drive, 65212, Columbia, MO, USA;0000 0001 2162 3504, grid.134936.a, Department of Biomedical Sciences, College of Veterinary Medicine, University of Missouri, 65212, Columbia, MO, USA;0000 0001 2162 3504, grid.134936.a, Department of Neurology, School of Medicine, University of Missouri, 65212, Columbia, MO, USA;0000 0001 2162 3504, grid.134936.a, Department of Bioengineering, University of Missouri, 65212, Columbia, MO, USA;

关键词: Dystrophin R16/17;    Blood perfusion;    Protein therapy;    Sarcolemmal nNOS;    Functional ischemia;    Dystrophinopathy;   

DOI  :  10.1186/s10020-019-0101-6

来源: publisher

【 摘 要 】

BackgroundDelocalization of neuronal nitric oxide synthase (nNOS) from the sarcolemma leads to functional muscle ischemia. This contributes to the pathogenesis in cachexia, aging and muscular dystrophy. Mutations in the gene encoding dystrophin result in Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD). In many BMD patients and DMD patients that have been converted to BMD by gene therapy, sarcolemmal nNOS is missing due to the lack of dystrophin nNOS-binding domain.MethodsDystrophin spectrin-like repeats 16 and 17 (R16/17) is the sarcolemmal nNOS localization domain. Here we explored whether R16/17 protein therapy can restore nNOS to the sarcolemma and prevent functional ischemia in transgenic mice which expressed an R16/17-deleted human micro-dystrophin gene in the dystrophic muscle. The palmitoylated R16/17.GFP fusion protein was conjugated to various cell-penetrating peptides and produced in the baculovirus-insect cell system. The best fusion protein was delivered to the transgenic mice and functional muscle ischemia was quantified.ResultsAmong five candidate cell-penetrating peptides, the mutant HIV trans-acting activator of transcription (TAT) protein transduction domain (mTAT) was the best in transferring the R16/17.GFP protein to the muscle. Systemic delivery of the mTAT.R16/17.GFP protein to micro-dystrophin transgenic mice successfully restored sarcolemmal nNOS without inducing T cell infiltration. More importantly, R16/17 protein therapy effectively prevented treadmill challenge-induced force loss and improved muscle perfusion during contraction.ConclusionsOur results suggest that R16/17 protein delivery is a highly promising therapy for muscle diseases involving sarcolemmal nNOS delocalizaton.

【 授权许可】

CC BY