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Apoptosis inhibitors and mini-agrin have additive benefits in congenital muscular dystrophy mice.

Meinen S, Lin S, Thurnherr R, Erb M, Meier T, Rüegg MA - EMBO Mol Med (2011)

Bottom Line: By combining mini-agrin with either transgenic Bcl2 expression or oral omigapil application, we show that the ameliorating effect of mini-agrin, which acts by restoring the mechanical stability of muscle fibres and, thereby, reduces muscle fibre breakdown and concomitant fibrosis, is complemented by apoptosis inhibitors, which prevent the loss of muscle fibres.Treatment of mice with both agents results in improved muscle regeneration and increased force.Our results show that the combination of mini-agrin and anti-apoptosis treatment has beneficial effects that are significantly bigger than the individual treatments and suggest that such a strategy might also be applicable to MDC1A patients.

View Article: PubMed Central - PubMed

Affiliation: Biozentrum, University of Basel, Switzerland. markus-a.ruegg@unibas.ch

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Effects of omigapil treatment in 12-week-old dyW/mag miceH & E staining of cross-sections from triceps brachii muscle of dyW/mag mice treated daily with 0.1 mg/kg omigapil (dyW/mag-omigapil) or vehicle (dyW/mag-vehicle).Blood CK levels expressed as percentage of control. Omigapil treatment in dyW/mag mice significantly lowers the CK levels (p = 0.023); N ≥ 6 ≤ 8.Muscle fibre size distribution of triceps brachii. Values represent relative numbers of fibres in a given diameter class (5 µm/class). Muscles from vehicle-treated dyW/mag mice contain more small-caliber fibres than muscles from age-matched dyW/mag-omigapil mice; N = 5.Mean fibre diameter. Omigapil treatment slightly but not significantly (p = 0.07) elevates the mean fibre size in dyW/mag muscles; N = 5.Muscle area (mm2) of mid-belly triceps cross-sections. Omigapil treatment tends to enlarge muscle area in dyW/mag mice (p = 0.06); N = 5.Total number of fibres per triceps cross-section. Omigapil treatment reduces muscle fibre loss in dyW/mag (p = 0.006) but does not completely normalize fibre numbers, as control muscle contain >6000 fibres (dashed line); N = 5.Weight gain normalized to the body weight at 15 days of age (onset of treatment). For each week, body weight data were averaged from daily measurements. Body weight at 15 days of age (starting body weight) was 5.55 ± 0.20 g for dyW/mag-omigapil mice and 6.88 ± 0.21 g for dyW/mag-vehicle mice. Body weight curve for vehicle-treated dyW/dyW mice (Erb et al, 2009) is shown for comparison (dotted line); N ≥ 9 ≤ 11.Locomotion within a 10 min observation period. Locomotive activity of dyW/mag-omigapil mice is significantly increased compared to vehicle-treated dyW/mag mice (p = 0.046). Locomotion of dyW/dyW (dashed line) and WT control mice (dotted line) are given for comparison; N ≥ 9 ≤ 11.Grip strength is expressed as time mice were able to hold on a vertical grid. Omigapil treatment significantly increases the grip strength in dyW/mag mice (p = 0.02). Grip strength of dyW/dyW mice is indicated (dashed line). WT control mice stay >180 s on the grid; N ≥ 9 ≤ 11.Kaplan–Meier cumulative survival plot. Daily treatment (0.1 mg/kg) with omigapil does not improve the survival probability of dyW/mag mice. All values are mean ± SEM. N indicates the animal number per experimental group. p-Values are Student's t-test (***p ≤ 0.001; **p ≤ 0.01; *p ≤ 0.05; n.s. p > 0.05). Scale bar = 50 µm.
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fig07: Effects of omigapil treatment in 12-week-old dyW/mag miceH & E staining of cross-sections from triceps brachii muscle of dyW/mag mice treated daily with 0.1 mg/kg omigapil (dyW/mag-omigapil) or vehicle (dyW/mag-vehicle).Blood CK levels expressed as percentage of control. Omigapil treatment in dyW/mag mice significantly lowers the CK levels (p = 0.023); N ≥ 6 ≤ 8.Muscle fibre size distribution of triceps brachii. Values represent relative numbers of fibres in a given diameter class (5 µm/class). Muscles from vehicle-treated dyW/mag mice contain more small-caliber fibres than muscles from age-matched dyW/mag-omigapil mice; N = 5.Mean fibre diameter. Omigapil treatment slightly but not significantly (p = 0.07) elevates the mean fibre size in dyW/mag muscles; N = 5.Muscle area (mm2) of mid-belly triceps cross-sections. Omigapil treatment tends to enlarge muscle area in dyW/mag mice (p = 0.06); N = 5.Total number of fibres per triceps cross-section. Omigapil treatment reduces muscle fibre loss in dyW/mag (p = 0.006) but does not completely normalize fibre numbers, as control muscle contain >6000 fibres (dashed line); N = 5.Weight gain normalized to the body weight at 15 days of age (onset of treatment). For each week, body weight data were averaged from daily measurements. Body weight at 15 days of age (starting body weight) was 5.55 ± 0.20 g for dyW/mag-omigapil mice and 6.88 ± 0.21 g for dyW/mag-vehicle mice. Body weight curve for vehicle-treated dyW/dyW mice (Erb et al, 2009) is shown for comparison (dotted line); N ≥ 9 ≤ 11.Locomotion within a 10 min observation period. Locomotive activity of dyW/mag-omigapil mice is significantly increased compared to vehicle-treated dyW/mag mice (p = 0.046). Locomotion of dyW/dyW (dashed line) and WT control mice (dotted line) are given for comparison; N ≥ 9 ≤ 11.Grip strength is expressed as time mice were able to hold on a vertical grid. Omigapil treatment significantly increases the grip strength in dyW/mag mice (p = 0.02). Grip strength of dyW/dyW mice is indicated (dashed line). WT control mice stay >180 s on the grid; N ≥ 9 ≤ 11.Kaplan–Meier cumulative survival plot. Daily treatment (0.1 mg/kg) with omigapil does not improve the survival probability of dyW/mag mice. All values are mean ± SEM. N indicates the animal number per experimental group. p-Values are Student's t-test (***p ≤ 0.001; **p ≤ 0.01; *p ≤ 0.05; n.s. p > 0.05). Scale bar = 50 µm.

Mentions: We have recently shown that oral application of the pharmacological apoptosis inhibitor omigapil to dyW/dyW mice ameliorates MDC1A pathology (Erb et al, 2009). Based on the finding that transgenic expression of Bcl2 increased the therapeutic effect of mini-agrin, we next tested whether the combination of mini-agrin and systemic application of omigapil would be of benefit. To this end, dyW/mag mice were treated with a daily dose of 0.1 mg/kg omigapil starting at the age of 15 days. The first week of treatment, omigapil was injected into the peritoneum followed by oral gavage after weaning. Mice were analysed at the age of 12 weeks. Visual inspection of H & E-stained cross-sections from triceps muscle did not reveal a striking difference between omigapil- (dyW/mag-omigapil) and vehicle- (dyW/mag-vehicle) treated dyW/mag mice (Fig 7A). Quantitative assessment of changes showed, however, lowered creatine kinase (CK) levels in the blood (Fig 7B), normalized muscle fibre size distribution (Fig 7C), a trend to increasing the mean fibre size (Fig 7D) and to enlarging muscle area in dyW/mag mice (Fig 7E). In addition, omigapil also reduced the muscle fibre loss in dyW/mag mice (Fig 7F). Importantly, many of the functional parameters were significantly improved by omigapil. Those included body weight gain (Fig 7G), locomotive activity in the open-field test (Fig 7H) and grip strength (Fig 7I). However, we could not detect a difference in the overall survival (Fig 7J). Together, treatment of dyW/mag mice with omigapil further improved several of the disease parameters but did not affect survival.


Apoptosis inhibitors and mini-agrin have additive benefits in congenital muscular dystrophy mice.

Meinen S, Lin S, Thurnherr R, Erb M, Meier T, Rüegg MA - EMBO Mol Med (2011)

Effects of omigapil treatment in 12-week-old dyW/mag miceH & E staining of cross-sections from triceps brachii muscle of dyW/mag mice treated daily with 0.1 mg/kg omigapil (dyW/mag-omigapil) or vehicle (dyW/mag-vehicle).Blood CK levels expressed as percentage of control. Omigapil treatment in dyW/mag mice significantly lowers the CK levels (p = 0.023); N ≥ 6 ≤ 8.Muscle fibre size distribution of triceps brachii. Values represent relative numbers of fibres in a given diameter class (5 µm/class). Muscles from vehicle-treated dyW/mag mice contain more small-caliber fibres than muscles from age-matched dyW/mag-omigapil mice; N = 5.Mean fibre diameter. Omigapil treatment slightly but not significantly (p = 0.07) elevates the mean fibre size in dyW/mag muscles; N = 5.Muscle area (mm2) of mid-belly triceps cross-sections. Omigapil treatment tends to enlarge muscle area in dyW/mag mice (p = 0.06); N = 5.Total number of fibres per triceps cross-section. Omigapil treatment reduces muscle fibre loss in dyW/mag (p = 0.006) but does not completely normalize fibre numbers, as control muscle contain >6000 fibres (dashed line); N = 5.Weight gain normalized to the body weight at 15 days of age (onset of treatment). For each week, body weight data were averaged from daily measurements. Body weight at 15 days of age (starting body weight) was 5.55 ± 0.20 g for dyW/mag-omigapil mice and 6.88 ± 0.21 g for dyW/mag-vehicle mice. Body weight curve for vehicle-treated dyW/dyW mice (Erb et al, 2009) is shown for comparison (dotted line); N ≥ 9 ≤ 11.Locomotion within a 10 min observation period. Locomotive activity of dyW/mag-omigapil mice is significantly increased compared to vehicle-treated dyW/mag mice (p = 0.046). Locomotion of dyW/dyW (dashed line) and WT control mice (dotted line) are given for comparison; N ≥ 9 ≤ 11.Grip strength is expressed as time mice were able to hold on a vertical grid. Omigapil treatment significantly increases the grip strength in dyW/mag mice (p = 0.02). Grip strength of dyW/dyW mice is indicated (dashed line). WT control mice stay >180 s on the grid; N ≥ 9 ≤ 11.Kaplan–Meier cumulative survival plot. Daily treatment (0.1 mg/kg) with omigapil does not improve the survival probability of dyW/mag mice. All values are mean ± SEM. N indicates the animal number per experimental group. p-Values are Student's t-test (***p ≤ 0.001; **p ≤ 0.01; *p ≤ 0.05; n.s. p > 0.05). Scale bar = 50 µm.
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fig07: Effects of omigapil treatment in 12-week-old dyW/mag miceH & E staining of cross-sections from triceps brachii muscle of dyW/mag mice treated daily with 0.1 mg/kg omigapil (dyW/mag-omigapil) or vehicle (dyW/mag-vehicle).Blood CK levels expressed as percentage of control. Omigapil treatment in dyW/mag mice significantly lowers the CK levels (p = 0.023); N ≥ 6 ≤ 8.Muscle fibre size distribution of triceps brachii. Values represent relative numbers of fibres in a given diameter class (5 µm/class). Muscles from vehicle-treated dyW/mag mice contain more small-caliber fibres than muscles from age-matched dyW/mag-omigapil mice; N = 5.Mean fibre diameter. Omigapil treatment slightly but not significantly (p = 0.07) elevates the mean fibre size in dyW/mag muscles; N = 5.Muscle area (mm2) of mid-belly triceps cross-sections. Omigapil treatment tends to enlarge muscle area in dyW/mag mice (p = 0.06); N = 5.Total number of fibres per triceps cross-section. Omigapil treatment reduces muscle fibre loss in dyW/mag (p = 0.006) but does not completely normalize fibre numbers, as control muscle contain >6000 fibres (dashed line); N = 5.Weight gain normalized to the body weight at 15 days of age (onset of treatment). For each week, body weight data were averaged from daily measurements. Body weight at 15 days of age (starting body weight) was 5.55 ± 0.20 g for dyW/mag-omigapil mice and 6.88 ± 0.21 g for dyW/mag-vehicle mice. Body weight curve for vehicle-treated dyW/dyW mice (Erb et al, 2009) is shown for comparison (dotted line); N ≥ 9 ≤ 11.Locomotion within a 10 min observation period. Locomotive activity of dyW/mag-omigapil mice is significantly increased compared to vehicle-treated dyW/mag mice (p = 0.046). Locomotion of dyW/dyW (dashed line) and WT control mice (dotted line) are given for comparison; N ≥ 9 ≤ 11.Grip strength is expressed as time mice were able to hold on a vertical grid. Omigapil treatment significantly increases the grip strength in dyW/mag mice (p = 0.02). Grip strength of dyW/dyW mice is indicated (dashed line). WT control mice stay >180 s on the grid; N ≥ 9 ≤ 11.Kaplan–Meier cumulative survival plot. Daily treatment (0.1 mg/kg) with omigapil does not improve the survival probability of dyW/mag mice. All values are mean ± SEM. N indicates the animal number per experimental group. p-Values are Student's t-test (***p ≤ 0.001; **p ≤ 0.01; *p ≤ 0.05; n.s. p > 0.05). Scale bar = 50 µm.
Mentions: We have recently shown that oral application of the pharmacological apoptosis inhibitor omigapil to dyW/dyW mice ameliorates MDC1A pathology (Erb et al, 2009). Based on the finding that transgenic expression of Bcl2 increased the therapeutic effect of mini-agrin, we next tested whether the combination of mini-agrin and systemic application of omigapil would be of benefit. To this end, dyW/mag mice were treated with a daily dose of 0.1 mg/kg omigapil starting at the age of 15 days. The first week of treatment, omigapil was injected into the peritoneum followed by oral gavage after weaning. Mice were analysed at the age of 12 weeks. Visual inspection of H & E-stained cross-sections from triceps muscle did not reveal a striking difference between omigapil- (dyW/mag-omigapil) and vehicle- (dyW/mag-vehicle) treated dyW/mag mice (Fig 7A). Quantitative assessment of changes showed, however, lowered creatine kinase (CK) levels in the blood (Fig 7B), normalized muscle fibre size distribution (Fig 7C), a trend to increasing the mean fibre size (Fig 7D) and to enlarging muscle area in dyW/mag mice (Fig 7E). In addition, omigapil also reduced the muscle fibre loss in dyW/mag mice (Fig 7F). Importantly, many of the functional parameters were significantly improved by omigapil. Those included body weight gain (Fig 7G), locomotive activity in the open-field test (Fig 7H) and grip strength (Fig 7I). However, we could not detect a difference in the overall survival (Fig 7J). Together, treatment of dyW/mag mice with omigapil further improved several of the disease parameters but did not affect survival.

Bottom Line: By combining mini-agrin with either transgenic Bcl2 expression or oral omigapil application, we show that the ameliorating effect of mini-agrin, which acts by restoring the mechanical stability of muscle fibres and, thereby, reduces muscle fibre breakdown and concomitant fibrosis, is complemented by apoptosis inhibitors, which prevent the loss of muscle fibres.Treatment of mice with both agents results in improved muscle regeneration and increased force.Our results show that the combination of mini-agrin and anti-apoptosis treatment has beneficial effects that are significantly bigger than the individual treatments and suggest that such a strategy might also be applicable to MDC1A patients.

View Article: PubMed Central - PubMed

Affiliation: Biozentrum, University of Basel, Switzerland. markus-a.ruegg@unibas.ch

Show MeSH
Related in: MedlinePlus