Proteomic profiling plays a decisive role in the elucidation of molecular signatures representative of a specific clinical context. MuStem cell based therapy represents a promising approach for clinical applications to cure Duchenne muscular dystrophy (DMD). To expand our previous studies collected in the clinically relevant DMD animal model, we decided to investigate the skeletal muscle proteome 4 months after systemic delivery of allogenic MuStem cells. Quantitative proteomics with isotope-coded protein labeling was used to compile quantitative changes in the protein expression profiles of muscle in transplanted Golden Retriever muscular dystrophy (GRMD) dogs as compared to Golden Retriever muscular dystrophy dogs. A total of 492 proteins were quantified, including 25 that were overrepresented and 46 that were underrepresented after MuStem cell transplantation. Interestingly, this study demonstrates that somatic stem cell therapy impacts on the structural integrity of the muscle fascicle by acting on fibers and its connections with the extracellular matrix. We also show that cell infusion promotes protective mechanisms against oxidative stress and favors the initial phase of muscle repair. This study allows us to identify putative candidates for tissue markers that might be of great value in objectively exploring the clinical benefits resulting from our cell-based therapy for DMD. All MS data have been deposited in the ProteomeXchange with identifier PXD001768 (http://proteomecentral.proteomexchange.org/dataset/PXD001768).
Essential for muscle fiber formation and hypertrophy, muscle stem cells, also called satellite cells, reside beneath the basal lamina of the muscle fiber. Satellite cells have been commonly localized by the expression of the Paired box 7 (Pax7) due to its specificity and the availability of antibodies in tetrapods. In fish, the identification of satellite cells remains difficult due to the lack of specific antibodies in most species. Based on the development of a highly sensitive in situ hybridization (RNAScope) for pax7, we showed that pax7+ cells were detected in the undifferentiated myogenic epithelium corresponding to the dermomyotome at day 14 post-fertilization. Then, from day 24, pax7+ cells gradually migrated into the deep myotome and were localized along the muscle fibers and reach their niche in satellite position of the fibres after hatching. Our results showed that 18 days after muscle injury, a large number of pax7+ cells accumulated at the wound site compared to the uninjured area. During the in vitro differentiation of satellite cells, the percentage of pax7+ cells decreased from 44% to 18% on day 7, and some differentiated cells still expressed pax7. Taken together, these results show the dynamic expression of pax7 genes and the follow-up of these muscle stem cells during the different situations of muscle fiber formation in trout.
1Background 2The dramatic increase in myotomal muscle mass in post-hatching fish is related to their ability to 3 lastingly produce new muscle fibres, a process termed hyperplasia. The molecular and cellular 4 mechanisms underlying fish muscle hyperplasia largely remain unknown. In this study, we aimed to 5 characterize intrinsic properties of myogenic cells originating from fish hyperplasic muscle. For this 6 purpose, we compared in situ proliferation, in vitro cell behavior and transcriptomic profile of 7 myogenic precursors originating from hyperplasic muscle of juvenile trout (JT) and from non-8 hyperplasic muscle of fasted juvenile trout (FJT) and adult trout (AT). 9 Results 10For the first time, we showed that myogenic precursors proliferate in hyperplasic muscle from JT as 11 shown by in vivo BrdU labeling. This proliferative rate was very low in AT and FJT muscle. 12 Transcriptiomic analysis revealed that myogenic cells from FJT and AT displayed close expression 13 profiles with only 64 differentially expressed genes (BH corrected p-val < 0.001). In contrast, 2623 14 differentially expressed genes were found between myogenic cells from JT and from both FJT and 15 AT. Functional categories related to translation, mitochondrial activity, cell cycle, and myogenic 16 differentiation were inferred from genes up regulated in JT compared to AT and FJT myogenic cells. 17 Conversely, Notch signaling pathway, that signs cell quiescence, was inferred from genes down 18 regulated in JT compared to FJT and AT. In line with our transcriptomic data, in vitro JT myogenic 19 precursors displayed higher proliferation and differentiation capacities than FJT and AT myogenic 20 precursors. 21 Conclusions 22The transcriptomic analysis and examination of cell behavior converge to support the view that 23 myogenic cells extracted from hyperplastic muscle of juvenile trout are intrinsically more potent to 24
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