A naturally occurring human minidysferlin protein repairs sarcolemmal lesions in a mouse model of dysferlinopathy
Authors: S√©bastien Courrier
Isabelle Richard
Marc Bartoli
Nicolas L√©vy
Nathalie Bourg-Alibert
Christophe P√©cheux
Nicolas Wein
V√©ronique Labelle
William Lostal
Karine Nguyen
Pierre Cau
Danielle DePetris
France Leturcq
Martin Krahn
Christophe Vial
Nathalie Streichenberger
Journal: Science Translational Medicine (2010)
Discipline: BIOLOGY & BIOCHEMISTRY
Keywords: Dysferlinopathies; DYSF; gene therapy; rAAV vectors; minidysferlin
Affiliations: Inserm UMR_S 910, G√©n√©tique M√©dicale et G√©nomique Fonctionnelle, Facult√© de M√©decine de Marseille, Universit√© de la M√©diterran√©e, 13005 Marseille, France.

Abstract:  Dysferlinopathies are autosomal recessive, progressive muscle dystrophies caused by mutations in DYSF, leading to a loss or a severe reduction of dysferlin, a key protein in sarcolemmal repair. Currently, no etiological treatment is available for patients affected with dysferlinopathy. As for other muscular dystrophies, gene therapy approaches based on recombinant adeno-associated virus (rAAV) vectors are promising options. However, because dysferlin messenger RNA is far above the natural packaging size of rAAV, full-length dysferlin gene transfer would be problematic. In a patient presenting with a late-onset moderate dysferlinopathy, we identified a large homozygous deletion, leading to the production of a natural "minidysferlin" protein. Using rAAV-mediated gene transfer into muscle, we demonstrated targeting of the minidysferlin to the muscle membrane and efficient repair of sarcolemmal lesions in a mouse model of dysferlinopathy. Thus, as previously demonstrated in the case of dystrophin, a deletion mutant of the dysferlin gene is also functional, suggesting that dysferlin's structure is modular. This minidysferlin protein could be used as part of a therapeutic strategy for patients affected with dysferlinopathies. 

Retracted: No

PubPeer comments:
 Figure 4. Induction of apoptosis by wogonoside and confirmation by Annexin V-FITC PI dual staining. SaOS-2 cells were treated with 0 (A, control), 5 u03bcM (B), 10 u03bcM (C), 25 u03bcM (D), and 75 u03bcM (E) of wogonoside for 48 h and analysed using FACS Calibur flow cytometer as described in u201cMaterials and Methodsu201d. Normal healthy, early apoptotic, late apoptotic and dead necrotic cell populations are shown as percentage of total cells in the quadrants Q3, Q4, Q2 and Q1, respectively. The heavy use of photoshop clone stamping indicates that these images are products of software rather than a flow cytometer.  , 
 #1  Figure 2. Effects of wogonoside on cell morphology change and percentage of viable SaOS-2 cells. SaOS-2 cells were cultured in DMEM + 10% FBS with different concentrations of wogonoside for 48 h. The cells were observed and photographed by phase-contrast microscopy (u00d7 200 magnification). A. Represents control (untreated cells), B-E. Represent 5, 10, 25 and 75 u00b5M dose of wogonoside. The number of viable cells decreases significantly as well as the cell morphology gets significantly altered as the wogonoside dose increases.  Not a lot of effort went into trying to make the different photoshop constructions look different.  ,