Proteomic profiling of antisense-induced exon skipping reveals reversal of
pathobiochemical abnormalities in dystrophic mdx diaphragm.
2009 Jan 8
Department of Biology, National University of Ireland,
Maynooth, Co. Kildare, Ireland.
The disintegration of the dystrophin-glycoprotein complex
represents the initial pathobiochemical insult in Duchenne muscular dystrophy.
However, secondary changes in signalling, energy metabolism and ion homeostasis
are probably the main factors that eventually cause progressive muscle wasting.
Thus, for the proper evaluation of novel therapeutic approaches, it is essential
to analyse the reversal of both primary and secondary abnormalities in treated
muscles. Antisense oligomer-mediated exon skipping promises functional
restoration of the primary deficiency in dystrophin. In this study, an
established phosphorodiamidate morpholino oligomer coupled to a cell-penetrating
peptide was employed for the specific removal of exon 23 in the mutated mouse
dystrophin gene transcript. Using DIGE analysis, we could show the reversal of
secondary pathobiochemical abnormalities in the dystrophic diaphragm following
exon-23 skipping. In analogy to the restoration of dystrophin, beta-dystroglycan
and neuronal nitric oxide synthase, the muscular dystrophy-associated
differential expression of calsequestrin, adenylate kinase, aldolase,
mitochondrial creatine kinase and cvHsp was reversed in treated muscle fibres.
Hence, the re-establishment of Dp427 coded by the transcript missing exon 23 has
counter-acted dystrophic alterations in Ca(2+)-handling, nucleotide metabolism,
bioenergetic pathways and cellular stress response. This clearly establishes the exon-skipping approach as a realistic
treatment strategy for diminishing diverse downstream alterations in
Link To Research Abstract
Link To Irish Times Article