Differential proteome analysis in col6a1-/- mouse model of gastrocnemious, tibialis and diaphragm muscles and effects of Ciclosporin A treatment

Ullrich Congenital Muscular Dystrophy (UCMD) and Bethlem Myopathy (BM) are congenital muscular dystrophies caused by abnormalities of Collagen VI due to COLVI gene mutations. The absence of Collagen VI has a major impact inside fibres by triggering a short circuit in the cell\u2019s energy generators, the mitochondria. This short circuit is caused by opening of a channel called the \u201cPermeability Transition Pore\u201d (PTP), which can be inhibited by the drug cyclosporin A (CsA). To better understand the molecular consequences of collagen VI absence in muscles, the proteome of functionally different and differentially affected muscles such as tibialis, diaphragm and gastrocnemius was investigated. Differential proteome was assessed by 2D-DIGE, De Cyder software analysis and MALDI Tof/Tof and ESI MS/MS. Results indicates that different categories of dysregulated proteins. A specific set of proteins was differentially regulated in diaphragm compared to tibialis and gastrocnemius suggesting that disease progression could be influenced by fiber type distribution, by muscle metabolism and by specific muscle function. The effects of CsA treatment were assessed in the same muscles adopting the same proteomic approach indicated above. Changes induced by CsA in muscle tissue were assessed by two-way ANOVA. A set of molecules, as putative targets of CsA treatment, were identified. A partial recovery of dysregulated proteins, more evident in gastronemious muscle, was observed according to muscle functional recovery. Moreover, CsA efficacy over time is not determined in this study, results highlight the relevance of the proposed approach to monitor effects of pharmacological treatments targeting muscle