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19 - (Am. J. Roentgenol., Jan 2008; 190: W8 - W12)Three-Point Technique of Fat Quantification of Muscle Tissue as a Marker of Disease Progression in Duchenne Muscular Dystrophy: Preliminary Study
OBJECTIVE. Clinical trials involving patients with Duchenne muscular dystrophy are hindered by the lack of suitable objective end points. The purpose of this study was to examine whether muscle lipid infiltration measured with the three-point Dixon MRI technique has value as a marker of disease severity.
SUBJECTS AND METHODS. Disease severity in nine boys (mean age, 8.6 ± 2.7 years) with Duchenne muscular dystrophy was determined with the functional ability scale of Brooke and associates. Functional scores were compared with strength measurements obtained by manual testing of muscles of the lower extremities, knee extensor strength measured with an isokinetic dynamometer, and muscle fat percentage in the quadriceps and hamstrings determined with the three-point Dixon MRI technique.
RESULTS. MRI measurements of fat infiltration had stronger correlation (p < 0.05) with functional grade than did measurements obtained with manual muscle testing (p = 0.07) or quantitative strength measured with the isokinetic dynamometer (p = 0.54). Muscle fat percentage did not correlate with strength measurements from manual or dynamometer muscle testing but increased with age in subjects with Duchenne muscular dystrophy.
CONCLUSION. Muscle adiposity values obtained with three-point Dixon MRI are accurate in assessment of disease severity in patients with Duchenne muscular dystrophy. Because they are not influenced by patient effort or examiner variability, these measurements are more objective and reproducible than measurements of muscle strength.
13 - (Chest. 132(6):1977-1986, December 2007) American College of Chest Physicians Consensus Statement on the Respiratory and Related Management of Patients With Duchenne Muscular Dystrophy Undergoing Anesthesia or Sedation
Birnkrant, David J.; Panitch, Howard B.; Benditt, Joshua O.; Boitano, Louis J. ; Carter, Edward R. MD, ; Cwik, Valerie A. ; Finder, Jonathan D. ; Iannaccone, Susan T. ; Jacobson, Lawrence E. ; Kohn, Gary L. ; Motoyama, Etsuro K. ; Moxley, Richard T. ; Schroth, Mary K. ; Sharma, Girish D. ; Sussman, Michael D. - USA
This statement on the management of patients with Duchenne muscular dystrophy (DMD) undergoing procedural sedation or general anesthesia represents the consensus opinion of a multidisciplinary panel convened under the auspices of the American College of Chest Physicians. Expert recommendations on this subject are needed for several reasons. First, patients with DMD have an increased risk of complications when they undergo sedation or general anesthesia. In addition, due to improved cardiopulmonary therapies, patients with DMD are experiencing an unprecedented duration of survival. As a result, it is more common for them to require procedures involving sedation or general anesthesia. The risks related to anesthesia and sedation for DMD patients include potentially fatal reactions to inhaled anesthetics and certain muscle relaxants, upper airway obstruction, hypoventilation, atelectasis, congestive heart failure, cardiac dysrhythmias, respiratory failure, and difficulty weaning from mechanical ventilation. This statement includes advice regarding the highly interrelated areas of respiratory, cardiac, GI, and anesthetic management of patients with DMD undergoing general anesthesia or procedural sedation. The statement is intended to aid clinicians involved in the care of patients with DMD and to be a resource for other stakeholders in this field, including patients and their families. It is an up-to-date summary of medical literature regarding this topic and identifies areas in need of future research.
13 - (Anaesthesia. 63(1):89-91, January 2008) Acute liver failure following therapeutic paracetamol administration in patients with muscular dystrophies.[Report]
Pearce, B.; Grant, I. S. - UK
Clinically significant liver damage in patients taking
therapeutic doses of paracetamol is very rare. We report two cases of fulminant
hepatic failure caused by therapeutic (4 g.day-1) paracetamol administration on
our Intensive Care Unit. Both patients had a
muscular dystrophy and presented
with a chest infection on a background of endstage neuromuscular
respiratory failure. We also noted one further similar case in the literature
and suggest a relationship between muscular
dystrophy and paracetamol-induced
hepatotoxicity. In this report we discuss in detail possible mechanisms that may
account for this apparent association, which include altered pharmacokinetics,
reduced glutathione stores, malnutrition and hypoxic injury.
12 - (Cell Stem Cell, December - 2007) Restoration of Human Dystrophin Following Transplantation of Exon-Skipping-Engineered DMD Patient Stem Cells into Dystrophic Mice
12 - PPMD Annual Connect Conference, Philadelphia, 12 - 14 July 2007 - Günter Scheuerbrandt report
8 - Cognitive and Psychological Profile of Males With Becker Muscular Dystrophy
NOVEMBER
14 - (47th Annual Meeting of American Society of Cell Biology, December 2007)
The Potential Influence of NF-kappaB Down Regulation on the Activity of the PI3/AkT Pathway in Dystrophic (mdx) Skeletal Muscle
B. Lee,1 G. W. Millman,1 E. C. Turin,1 A. Samadi,2 C. G. Carlson1; 1Physiology, A.T. Still University of Health Sciences, Kirksville, MO, 2Biochemistry, A.T. Still University of Health Sciences, Kirksville, MO
Recent evidence indicating that treatments to reduce the activity of the NF-kappaB pathway may be useful in treating Duchenne and Becker muscular dystrophy (Carlson et al., Neurobiology of Disease, 20 (3):719-730, 2005; Messina et al., Experimental Neurology 198:234-241, 2006; Acharyya et al., J. Clin. Invest. 117, 889-901, 2007) provides the rationale for ongoing experiments which are directed at identifying specific phenotypic endpoints for assessing drug efficacy using the mdx mouse model for Duchenne muscular dystrophy. Consideration of the central importance of the PI3kinase/Akt pathway in promoting the growth and regeneration of skeletal muscle, however, suggests that a combined treatment to reduce the activity of the NF-kappaB pathway and increase PI3kinase/Akt activation may be an appropriate therapy for this group of diseases. In order to investigate this possibility, we have begun to examine the status of the PI3kinase/Akt pathway in costal diaphragm cytosolic extracts obtained from mdx mice treated either acutely or chronically with two inhibitors of the NF-kappaB pathway, pyrrolidine dithiocarbamate (PDTC; 50 mg/kg) and sulfasalazine (SS; 100 mg/kg). Western blots of phosphorylated and total Akt indicate that a single injection of either of these two agents does not modify the activity of the PI3kinase/Akt pathway. Initial experiments suggest that daily treatment of mdx mice with PDTC for a period of at least 30 days may activate the PI3kinase/Akt pathway, but that corresponding treatment with sulfasalazine has no effect on the proportion of phosphorylated Akt. Although it is currently unclear whether chronic treatment with these established NF-kappaB inhibitors effectively down regulates nuclear NF-kappaB activation in dystrophic skeletal muscle, these preliminary results suggest that chronic treatment with NF-kappaB inhibitors that reduce IkappaB-alpha kinase activity (IKK) may be used in conjunction with PI3kinase/Akt activation to judiciously enhance muscle regeneration in dystrophic skeletal muscle.
Evaluation of the Effectiveness of a Mast Cell Degranulation Inhibitor in Dystrophic mdx Mice
D. D. Araujo,1,2 F. S. M. Pires,2 T. T. Fuzisaki,2 J. C. S. Bizario,2 M. R. Costa1,2; 1Cellular Molecular Biology, University of São Paulo, Ribeirão Preto, Brazil, 2Medicine School, University of Ribeirão Preto, Ribeirão Preto, Brazil
Duchenne muscular dystrophy (DMD) is characterized by progressive muscle degeneration which results from the absence of dystrophin. The dystrophin deficient mdx mouse, the most widely used animal model for DMD, shows milder phenotype that can be intensified by compulsory physical activity. Inflammatory response and immune cells contribute to muscle degeneration and/or regeneration. Mast cells are recruited to the injury sites and liberate mediators that increase collagen production, attract other inflammatory cells and intensify tissue damage. Olopatadine, an H1-histamine antagonist and a mast cell degranulation inhibitor, decreases Ca2+ influx and the liberation of histamine, tryptase, leukotrienes and prostaglandins. In this study, we evaluated the effect of 10 mg/kg/day olopatadine in male mdx mice submitted to physical activity (20 minutes of treadmill running at 20 cm/s, 3 times a week, during 5 weeks). To characterize the experimental groups (treated and untreated) before (4 week old mdx) and after (9 week old mdx) of the physical activity program, we used the following parameters: i) measurement of body weight and serum creatine kinase; ii) qualitative and quantitative histopathological analyses of gastrocnemius and diaphragm (Evans Blue Dye infiltration and H&E staining to evaluate myofibers morphology, the presence of inflammatory infiltrates, peripherally and centrally nucleated myofibers, and grouped regenerating myofibers). The results showed that the 9 week old olopatadine treated mice presented significant reduction in the CK levels and amelioration of the muscular condition. We detected significant reduction in the number of myofibers with sarcolemma injury but no difference in the number of centrally nucleated myofibers. The analysis of the individual values suggests a trend to reduction in the inflammatory infiltrate area and in the degeneration/regeneration ratio. These data suggest that the inhibition of mast cell is beneficial to retard the dystrophinopathy progress and can be considered as an option to clinical trials for DMD.
13 - Kiwi company's medical breakthrough
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10 - (Journal of the Neurological Sciences, 2007) Connective tissue growth factor is overexpressed in muscles of human muscular dystrophy
Guilian Sun, Kazuhiro Haginoya, Yanling Wu , Yoko Chiba , Tohru Nakanishi ,Akira Onuma , Yuko Sato e, Masaharu Takigawa , Kazuie Iinuma , Shigeru Tsuchiya - Japan
The detailed process of how dystrophic muscles are replaced by fibrotic tissues is unknown. In the present study, the immunolocalization and mRNA expression of connective tissue growth factor (CTGF) in muscles from normal and dystrophic human muscles were examined with the goal of elucidating the pathophysiological function of CTGF in muscular dystrophy. Biopsies of frozen muscle from patients with Duchenne muscular dystrophy (DMD), Becker muscular dystrophy, congenital muscular dystrophy, spinal muscular atrophy, congenital myopathy were analyzed using anti-CTGF polyclonal antibody. Reverse transcription-polymerase chain reaction (RT-PCR) was also performed to evaluate the expression of CTGF mRNA in dystrophic muscles. In normal muscle, neuromuscular junctions and vessels were CTGF-immunopositive, which suggests a physiological role for CTGF in these sites. In dystrophic muscle, CTGF immunoreactivity was localized to muscle fiber basal lamina, regenerating fibers, and the interstitium. Triple immunolabeling revealed that activated fibroblasts were immunopositive for CTGF and transforming growth factor-beta1 (TGF-beta1). RT-PCR analysis revealed increased levels of CTGF mRNA in the muscles of DMD patients. Co-localization of TGF-beta1 and CTGF in activated fibroblasts suggests that CTGF expression is regulated by TGF-beta1 through a paracrine/autocrine mechanism. In conclusion, TGF-beta1-CTGF pathway may play a role in the fibrosis that is commonly observed in muscular dystrophy.
8 - (Prostaglandins, Leukotrienes and Essential Fatty Acids, 2007) Effects of inhibitors of the arachidonic acid cascade on primary muscle culture from a Duchenne muscular dystrophy patient
I. Gáti O., Danielsson, T. Betmark, J. Ernerudh, K. Öllinger and N. Dizdar - Sweden
The aim of this study was to elucidate the mechanisms of action for potential targets of therapeutic intervention related to the arachidonic acid cascade in muscular dystrophy. Primary cultures from a Duchenne patient were used to study the expression of dystrophin-1, utrophin, desmin, neonatal myosin heavy chain (MHCn) and Bcl-2 during inhibition of phospholipase A2 (PLA2), cyclooxygenase (COX) and lipoxygenase (LOX). Hypo-osmotic treatment was applied in order to trigger Ca2+ influx and PLA2 activity. Inhibition of PLA2 and LOX with prednisolone and nordihydroguaiaretic acid (NDGA) caused a semi-quantitative increase of utrophin and Bcl-2-, and a dose-dependent, quantitative increase of desmin expression, an effect that was augmented by hypo-osmotic treatment. Our results indicate that LOX inhibitors, similarly to corticosteroids, can be beneficial in the treatment of muscular dystrophies
6 - (American Heart Association Annual Meeting, 2007) 1) Minor Myocardial Damage and LV Dysfunction in Patients with Duchenne Muscular Dystrophy -The Preventive Efficacy of Carvedilol on Plasma Cardiac Troponin I-
Background: Cardiac dysfunction is one of the major prognostic factors in patients with Duchenne muscular dystrophy (DMD). Minor myocardial damage assessed by plasma cardiac troponin I (cTnI) is often observed in patients with DMD. However, it is unclear that how the minor myocardial damage occurs to which patient with DMD and how it relates to LV dysfunction. Therefore, we assessed the hypothesis that minor myocardial damage is associated with LV dysfunction, the evaluation of plasma cTnI helps the prediction of LV functional deterioration, and carvedilol prevents elevation of plasma cTnI in patients with DMD.
Methods: Plasma cTnI were repeatedly (every 3 months) measured for 2 years and LV function was assessed by echocardiography in 58 patients with DMD. Carvedilol (2.5–5 mg/day) was orally administered for a year to the patients who have shown plasma cTnI elevation (positive cTnI, cut off 0.06ng/mL).
Results: There were 3 differential groups regarding the progression rate of LV systolic dysfunction, i.e. rapid (19% of total, LVEF < 50% in their 10th), slow (50% of total, LVEF > 50% in their 20 –30th), and unchanged group (31% of total, LVEF < 50% in their 20 –30th). The episode of positive cTnI was observed in 27 (46%) of total patients with DMD. LVEF was lower in patients with positive cTnI than that with negative cTnI (42 ± 2 vs. 52 ± 2%, p < 0.05). Positive cTnI was observed in all patients in rapid group, 84% of patients in slow group, and only 6% of patients in unchanged group. Fourteen differential dystrophin gene mutations were recognized in 48 patients but they were not associated with those 3 differential groups or patients with positive cTnI. Administration of carvedilol in 13 patients (LVEF 40 ± 3) decreased the cTnI detection rate (from 44 ± 5% to 26 ± 10%, p < 0.05), while it was unchanged in 14 patients (LVEF 41 ± 3) without carvedilol treatment (from 44 ± 7% to 39 ± 6%) during same observation period.
Conclusions: The elevation of plasma cTnI was associated with LV systolic dysfunction in patients with DMD. The prediction of LV dysfunction in patients with DMD may be feasible with combinations of age, LVEF, and plasma cTnI elevation. Carvedilol could be a new therapeutic strategy to prevent minor myocardial damage in patients with DMD.
Background: Delta-Sarcoglycan (SGCD) is a member of the dystrophin-associated glycoprotein complex. Mutations may result in isolated dilated cardiomyopathy or limb-girdle muscle dystrophy 2F with variable cardiac involvement. A potential gene therapeutic approach requires an efficient myocardial gene transfer. AAV vectors are suitable for cardiac gene transfer. Novel AAV serotypes such as AAV-8 or -9 even enable a systemic gene transfer in rodents. Aim of our study was to establish a vector for efficient systemic cardiac gene transfer and its subsequent use to prevent heart failure by expressing the SGCD cDNA in SGCD knock out mice.
Material and Methods: 1011 genomic particles of AAV-8 and -9 vectors harboring a luciferase reporter gene under control of the CMV-enhanced myosin light chain promoter were intravenously injected into adult NMRI mice (n=6 and n=10, respectively). After 4 weeks, reporter activities were determined in representative organs. Analogous, 2x1011 AAV-9 SGCD and AAV-9 EGFP vectors were intravenously injected in SGCD knock out mice (n=9 and n=7, respectively). Cardiac function (fraction of shortening) was assed by echocardiography at the beginning and the end of the study after 6 months. Running distances were measured using voluntary wheel running.
Results: Reporter gene transfer with AAV-9 vectors resulted in increased cardiac luciferase activities compared to AAV-8 (3.8x108 ±4.4x108 relative light units [RLU]/mg protein versus 0.10x108 ±0.08x108 RLU/mg protein, p=0.05) with increased specificity. In comparison to EGFP controls, AAV-9 delta-sarcoglycan gene transfer resulted in a significant increase in running distance and a significant decrease in decline in fraction of shortening (67.6% to 65.0% versus 69.9% to 58.1%; p=0.04). Immunohistochemical analyses in AAV-9 SGCD treated knock-out mice revealed a transmural reconstitution of more than 90% of cardiomyocytes.
Conclusion: Intravenous injections of AAV-9 SGCD vectors enable an efficient transmural cardiac gene transfer in adult mice and prevent deterioration of left ventricular function in SGCD knock out mice. Therefore, this vector system may be suitable for validation of novel targets for cardiac diseases
BACKGROUND: Duchenne Muscular Dystrophy (DMD), an x-linked myopathy, is characterized by progressive cardiac dysfunction and myocardial fibrosis late in the disease process. We hypothesized that left ventricular myocardial peak circumferential strain (cc) abnormalities would exist in DMD prior to global systolic functional abnormalities regardless of age or left ventricular ejection fraction (EF).
METHODS: We evaluated 33 DMD males undergoing clinical cardiac MRI. Comparison was made to 13 age-matched healthy volunteers. Standard imaging included short-axis cine stack, cine myocardial tagging and myocardial delayed enhancement (MDE). Cardiac functional analysis was performed using Medis MASS software. Tagged images of only the mid-LV were analyzed using HARP software. For analysis, DMD males were subdivided by age: < or > 10 years. All subjects < 10 years had normal EF (>55%); none exhibited MDE. Subjects >10 years were divided by MDE status and EF (>55% or <55%). Student’s t-test was used to test for differences between groups.
RESULTS: DMD subjects with normal EF had abnormal cc at an early age (<10 years) compared to control subjects (p< 0.05). DMD subjects >10 years with normal EF had further decline in =cc compared to younger DMD boys (p<0.05). There was further decline with age in =cc in subjects with lower EF (p<0.05) without MDE. In the oldest males studied with MDE and reduced EF, cc was even lower (figure).
CONCLUSIONS: Myocardial strain abnormalities are prevalent in the early stages of DMD existing in the setting of normal EF and progressing with advances in age. Such strain analysis may provide an early marker to guide the efficacy of preventive or gene therapy in DMD.
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2 - (Journal of Electromyography and Kinesiology, 2007) Evaluation of plantar flexion contracture contribution during the gait of children with Duchenne muscular dystrophy
Nathaly Gaudreailt, Denis Gravel and Sylvie Nadeau - Canada
Because of extensor weakness, children with Duchenne muscular dystrophy (DMD) maintain internal flexion moments at the joints of the lower extremities when they walk. We believe that at the ankle, the plantar flexion moments caused by contractures may contribute significantly to the production of the net ankle flexion moment during the gait in these children. The goal of the present study is to quantify ankle plantar flexion passive moments that may be associated with the presence of flexion contractures and to estimate their contribution to the net moment during the gait of children with DMD. Kinematic and kinetic parameters were collected during gait of eleven subjects with DMD. Ankle plantar flexion passive moments were also measured experimentally during the same session. Fourteen control children participated in the study in order to have normal reference values. The presence of ankle plantar flexion contractures in children with DMD was reflected by a rigidity coefficient obtained at a common moment of −7 Nm that was higher for these children (0.75 Nm/° vs. 0.48 Nm/°; p < 0.05). The relative passive moment contribution to the net plantar flexion moments was higher for the children with DMD at the end of the lengthening phase of the plantar flexors (25% vs. 18%; p < 0.05). We believe that the passive moments can compensate for the presence of progressive muscle weakness in the children with DMD and help these children with gait.
2 - (Am. J. Pathol., Nov 2007; 171: 1576 - 1587) Transgenic Overexpression of ADAM12 Suppresses Muscle Regeneration and Aggravates Dystrophy in Aged mdx Mice
Muscular dystrophies are characterized by insufficient restoration and gradual replacement of the skeletal muscle by fat and connective tissue. ADAM12 has previously been shown to alleviate the pathology of young dystrophin-deficient mdx mice, a model for Duchenne muscular dystrophy. The observed effect of ADAM12 was suggested to be mediated via a membrane-stabilizing up-regulation of utrophin, 7B integrin, and dystroglycans. Ectopic ADAM12 expression in normal mouse skeletal muscle also improved regeneration after freeze injury, presumably by the same mechanism. Hence, it was suggested that ADAM12 could be a candidate for nonreplacement gene therapy of Duchenne muscular dystrophy. We therefore evaluated the long-term effect of ADAM12 overexpression in muscle. Surprisingly, we observed loss of skeletal muscle and accelerated fibrosis and adipogenesis in 1-year-old mdx mice transgenically overexpressing ADAM12 (ADAM12+/mdx mice), even though their utrophin levels were mildly elevated compared with age-matched controls. Thus, membrane stabilization was not sufficient to provide protection during prolonged disease. Consequently, we reinvestigated skeletal muscle regeneration in ADAM12 transgenic mice (ADAM12+) after a knife cut lesion and observed that the regeneration process was significantly impaired. ADAM12 seemed to inhibit the satellite cell response and delay myoblast differentiation. These results discourage long-term therapeutic use of ADAM12. They also point to impaired regeneration as a possible factor in development of muscular dystrophy.
OCTOBER
27 - (JCB, 2007;179(2):305) Necdin mediates skeletal muscle regeneration by promoting myoblast survival and differentiation
Daniela Deponti, Stéphanie François, Silvia Baesso, Clara Sciorati, Anna Innocenzi, Vania Broccoli, Françoise Muscatelli, Raffaella Meneveri, Emilio Clementi, Giulio Cossu, and Silvia Brunelli - Italy
Regeneration of muscle fibers that are lost during pathological muscle degeneration or after injuries is sustained by the production of new myofi bers. An important cell type involved in muscle regeneration is the satellite cell. Necdin is a protein expressed in satellite cell–derived myogenic precursors during perinatal growth. However, its function in myogenesis is not known. We compare transgenic mice that overexpress necdin in skeletal muscle with both wild-type and necdin null mice. After muscle injury the necdin null mice show a considerable defect in muscle healing, whereas mice that overexpress necdin show a substantial increase in myofiber regeneration. We also find that in muscle, necdin increases myogenin expression, accelerates differentiation, and counteracts myoblast apoptosis. Collectively, these data clarify the function and mechanism of necdin in skeletal muscle and show the importance of necdin in muscle regeneration.
26 - (Circulation Research, October 2007) Twenty-One-Month-Old Carrier Mice by Mosaic Dystrophin Expression or Complementary Dystrophin/Utrophin Expression
Brian Bostick ;
A cure for dystrophin-deficient muscular dystrophy requires treating both skeletal muscle and the heart. Whereas mosaic dystrophin expression has been shown to protect skeletal muscle, controversy exists over whether mosaic expression is protective in the heart. We have shown recently that mosaic dystrophin expression prevents stress-induced heart damage in young carrier mice. Although an interesting finding, the clinical relevance remains to be established because young dystrophin-null mdx mice do not have heart disease. On the other hand, heart failure has been reported in human carriers. To resolve this mouse/human discrepancy, we evaluated the cardiac phenotype in 21-month-old mdx, carrier, and normal mice. We found dilated cardiomyopathy in old mdx mice but not in age-matched carrier mice. All anatomical parameters and physiological assay results (ECG and closed-chest Millar catheter) were within the normal range in old carrier mice. Focal myocardial inflammation was found in a small fraction of old carrier mice, but it had no major impact on heart function. Dobutamine stress revealed a near normal hemodynamic profile except for a marginal reduction in systolic pressure in old carrier mice. Immunostaining and Western blot showed dystrophin expression in 50% cardiomyocytes in old carrier mice. Interestingly, utrophin was upregulated in dystrophin-negative heart cells in carrier mice. In summary, we have provided the first clear-cut evidence that dilated cardiomyopathy in old mdx mice was prevented by mosaic dystrophin expression or complementary dystrophin/utrophin expression. Our results raise the hope for ameliorating dystrophic cardiomyopathy through partial gene and/or cell therapy.
20 - (Chest Meeting Abstracts, Oct 2007; 132: 607c - 608) INCIDENCE AND OUTCOMES OF CARDIOPULMONARY RESUSCITATION IN PATIENTS WITH END-STAGE DUCHENNE MUSCULAR DYSTROPHY
Garey Noritz David J. Birnkrant - USA
PURPOSE: Although technological cardiopulmonary therapies, such as noninvasive ventilation (NIV), have increased life span for patients (pts) with Duchenne muscular dystrophy (DMD), the disease remains fatal. Pts with end-stage DMD who forgo “do not resuscitate” directives may be partially resuscitated after near-fatal events, exposing them to the risk of brain damage. PURPOSE: To assess the frequency of attempted resuscitation among pts with DMD in our clinic who experienced cardiopulmonary arrest, and to describe their outcomes.
METHODS: Retrospective chart review.
RESULTS: Ten of our pts with DMD died during the time period 9/1/03-4/1/07. Mean age at death _/- SD: 20.7 _/- 2.7 years. Three of the 10 pts had unmeasurable vital capacities and were dependent on NIV 24 hrs/day. Mean vital capacity of the remaining 7 pts: 0.57 _/- 0.53 liters. All pts were prescribed daily NIV but 4 pts used NIV infrequently. All pts had cardiomyopathy. Four pts were in congestive heart failure. Two pts had cardiac defibrillators. After cardiopulmonary arrest, five of the 10 pts underwent resuscitation attempts. No pt was successfully resuscitated. Four of the 5 pts died immediately. One pt was partially resuscitated, experienced brain damage, and died after being taken off the ventilator by his family.
CONCLUSION: A significant proportion of our pts with end-stage DMD who experienced cardiopulmonary arrest underwent attempted resuscitation. None of the resuscitation attempts was successful. Lack of “do not resuscitate” advance directives exposed these pts to the risk of partial resuscitation and brain damage.
CLINICAL IMPLICATIONS: Pts with advanced DMD are increasingly cared for by cardiologists, pulmonologists, and intensivists, whose focus may be technologically oriented. We speculate that use of technological therapies like assisted ventilation and defibrillators among such pts may produce an unrealistically optimistic view of their prognosis, causing resistance to advance “do not resuscitate” directives, even among pts with end-stage disease. Studies are needed regarding how to reconcile palliative and technological therapies to optimize end-of-life care among pts with severe DMD.
18 - Distinctive patterns of microRNA expression in primary muscular disorders
17 - (Neurology, October 2007) Albuterol increases lean body mass in ambulatory boys with Duchenne or Becker muscular dystrophy
C. L. Skura,
Background: Albuterol is a beta-2 agonist that has been demonstrated to increase muscle strength in studies in animals and humans. Based on a pilot study of extended-release albuterol Repetabs in children with dystrophinopathies, the authors conducted a randomized, double-blind, placebo-controlled study with a crossover design.
Methods: Fourteen boys with Duchenne or Becker muscular dystrophy, 6 to 11 years old, completed two treatment periods (albuterol and placebo), 12 weeks each, separated by a 12-week washout period. As the albuterol Repetab formulation was no longer available, an alternate extended release albuterol was used (Volmax, 12 mg per day). Outcome measurements included 1) lean body mass, 2) fat mass, 3) isometric knee extensor and flexor moments, 4) manual muscle testing, and 5) timed functional tests.
Results: Lean body mass was significantly higher for subjects following albuterol treatment compared to placebo treatment, while fat mass was significantly lower. No differences were found in isometric knee moments or manual muscle tests. Time to run/walk 30 feet was improved following albuterol.
Conclusions: Short-term treatment with extended release albuterol may increase lean body mass, decrease fat mass, and improve functional measures in patients with dystrophinopathies. However, the significant change in strength of specific muscle groups found in the pilot study was not observed in the present study. These findings may be attributed to differences in the drug release and kinetics between Repetab and Volmax formulations as they affect the concentration of available beta-2 receptors on the muscle cell surface differently.
13 - Safety and Efficacy Study of Antisense Oligonucleotides in Duchenne Muscular Dystrophy
12 - (European Journal of Paediatric Neurology 11 (2007) 337-40) CINRG pilot trial of oxatomide in steroid-naıve Duchenne muscular dystrophy
Gunnar M. Buyse,, Nathalie Goemans, Erik Henricson, Alejandro Jara, Marleen van den Hauwe, Robert Leshner, Julaine M. Florence, Jill E. Mayhew, Diana M. Escolar - USA
The authors report a pilot open-label two-center therapeutic trial of oxatomide in 14 steroid-naive DMD boys aged 5–10 years. Comparison of linear evolutions between 3 months medication-free lead-in periods and 6 months treatment periods showed no significant differences in quantitative (QMT) and manual (MMT) measurements of muscle strength and timed functional tests. A modest mitigation of strength deterioration over time cannot be excluded.
12 - (Heart, October - 2007) Life expectancy and death from cardiomyopathy amongst carriers of Duchenne and Becker muscular dystrophy in Scotland
Objectives To assess life expectancy and cardiovascular mortality in carriers of Duchenne and Becker Muscular Dystrophy.
Design Family pedigrees of individuals affected with these conditions, held by the 4 genetics centres in Scotland, were examined to identity a cohort of definite carriers. Electronic death registration data, held by the General Register Office for Scotland were used to identify death certificates of carriers who had died, to obtain age at death and cause of death. Survival and mortality data were obtained for the general population for comparison.
Patients 397 definite carriers in 202 pedigrees were identified from which 94 deaths were identified by record linkage to death certificates.
Main outcome measures Observed numbers surviving to certain ages and numbers dying of cardiac causes were compared with expected numbers calculated from general population data. Results: There were no significant differences between observed and expected numbers surviving to ages 40-90. The standardised mortality ratio for the 371 carriers alive in 1974 was 0.53 (95% confidence interval 0.32-0.82).
Conclusions Whereas female carriers may have clinical features of cardiomyopathy, this study does not suggest that this is associated with reduced life expectancy or increased risk of cardiac death. Routine cardiac surveillance of obligate carriers is therefore probably unnecessary.
9 - (Paediatrics and Child Health, Volume 17, Issue 10, October 2007, Pages 419-420) Bone profile and vitamin D level in children with Duchenne muscular dystrophy (DMD)
M Sundaram, S Spinty - Alder Hey Childrens Hospital, Liverpool
Introduction: Approximately 1500 children are affected by DMD in the UK at any one time. Patients with DMD are at increased risk of long-bone and vertebral fractures. Reduced physical activity, a calcium-poor diet and low vitamin D levels are risk factors for reduced bone mineral density. Low vitamin D levels increase the risk of osteopenia. Inadequate nutritional intake of the precursor vitamin D2 and lack of sun exposure to aid conversion to the active form vitamin D3 in the skin lowers vitamin D levels. Sun exposure is often reduced in wheelchair-dependent patients with DMD. Steroids have been shown to slow down disease progression in patients with DMD. It is now recommended that all children with DMD should be offered steroids at a time when their functional abilities plateau. Osteoporosis is a side-effect of steroid treatment and further increases the risk of bone fractures. We set out to evaluate bone profiles (calcium, phosphate and alkaline phosphate) and vitamin D levels in children and adolescents with DMD, whose samples were taken at Alder Hey Hospital. Methods: Bone profiles and vitamin D2 and D3 levels were reviewed in 46 patients with DMD. Results: None of the patients studied showed clinical or biochemical evidence of rickets. Vitamin D3 levels were low for the seasonal variation in 17 out of 46 (36.9%) patients – 6 out of the 23 (26%) ambulant patients and 11 out of the 23 non-ambulant (47.8%) patients were vitamin D deficient. A number of children were vitamin D deficient at diagnosis. The mean age of the patients with low vitamin D levels was 11.6 years (range 7–17 years). Conclusions: Vitamin D level should be checked routinely at diagnosis and at regular intervals in all children with DMD. Deficiency should be appropriately treated to optimise bone health and reduce the risk of fractures.
5 - Autologous transplantation of muscle-derived CD133+ stem cells in Duchenne muscle patients
4 - Use of Weekly Alendronate to Treat Osteoporosis in Boys with Muscular Dystrophy
4 - (NEUROSCI BIOBEHAV REV, 2007) Duchenne muscular dystrophy: A cerebellar disorder?
CYRULNIK, S.C., and V.J. Hinton - USA
Duchenne muscular dystrophy (DMD) is a genetic disorder that is often associated with cognitive deficits. These cognitive deficits have been linked to the absence of dystrophin, a protein product which is normally found in multiple tissues throughout the body. In the current paper, we argue that it is the absence of dystrophin in the cerebellum that is responsible for the cognitive deficits observed. We begin by reviewing data that document structural and functional abnormalities in the brains of individuals with DMD and mdx mice. We briefly review the cognitive deficits associated with DMD, and then present neuroimaging and neuropsychological evidence to indicate that the cerebellum is involved in the same aspects of cognition that are impaired in children with DMD. It is our contention that brain pathways in the cerebellum (e.g., cerebro-cerebellar loops) which develop without dystrophin may result in altered brain function presenting as cognitive deficits in DMD.
SEPTEMBER
24 - (Faseb J, 2007) Transgenic expression of a myostatin inhibitor derived from follistatin increases skeletal muscle mass and ameliorates dystrophic pathology in mdx mice
Myostatin is a potent negative regulator of skeletal muscle growth. Therefore, myostatin inhibition offers a novel therapeutic strategy for muscular dystrophy by restoring skeletal muscle mass and suppressing the progression of muscle degeneration. The known myostatin inhibitors include myostatin propeptide, follistatin, follistatin-related proteins, and myostatin antibodies. Although follistatin shows potent myostatin-inhibiting activities, it also acts as an efficient inhibitor of activins. Because activins are involved in multiple functions in various organs, their blockade by follistatin would affect multiple tissues other than skeletal muscles. In the present study, we report the characterization of a myostatin inhibitor derived from follistatin, which does not affect activin signaling. The dissociation constants (Kd) of follistatin to activin and myostatin are 1.72 nM and 12.3 nM, respectively. By contrast, the dissociation constants (Kd) of a follistatin-derived myostatin inhibitor, designated FS I-I, to activin and myostatin are 64.3 µM and 46.8 nM, respectively. Transgenic mice expressing FS I-I, under the control of a skeletal muscle-specific promoter showed increased skeletal muscle mass and strength. Hyperplasia and hypertrophy were both observed. We crossed FS I-I transgenic mice with mdx mice, a model for Duchenne muscular dystrophy. Notably, the skeletal muscles in the mdx/FS I-I mice showed enlargement and reduced cell infiltration. Muscle strength is also recovered in the mdx/FS I-I mice. These results indicate that myostatin blockade by FS I-I has a therapeutic potential for muscular dystrophy
23 - (2007 Annual Meeting of The American Society of Human Genetics) Antisense-mediated exon 51 skipping restores local dystrophin expression in muscle of Duchenne muscular dystrophy patients.
A. Aartsma-Rus1, J.J.G.M. Verschuuren2,
A.A.M. Janson3, G. Platenburg3, G-J.B. van Ommen1,
J.C.T. van Deutekom1,3 1) Department of Human Genetics, Leiden
University Medical Center, Leiden, the Netherlands; 2) Department of Neurology,
Leiden University Medical Center, Leiden, the Netherlands; 3) Prosensa B.V.
Leiden, the Netherlands.
Antisense-mediated reading frame restoration is currently one of the most
promising therapeutic approaches for Duchenne muscular dystrophy (DMD). In this
approach, antisense oligoribonucleotides (AONs) induce specific exon skipping
during pre-mRNA splicing. They have been successful in repairing the disrupted
open reading frame accompanied by the generation of internally deleted,
partially functional Becker-like dystrophins. Proof of concept has been achieved
in cultured muscle cells from patients, as well as in the mdx mouse model.
As an essential step towards broad clinical studies and future applications, we
here evaluated the effect of a single, intramuscular dose of DMD AON PRO051.
Four DMD patients with different mutations were included on basis of eligible
mutation, adequate condition of the target muscle, and positive in vitro
PRO051 skip-response. A dose of 0.8 mg PRO051, without any excipient, was
injected locally into tibialis anterior muscle and a biopsy was taken after 4
weeks. Exon 51 skipping on RNA level and restoration of dystrophin expression
was confirmed for each patient, as demonstrated by RT-PCR, immunohistochemical
and western blot analyses. Dystrophin levels were ~10% of wild type levels,
except for one patient who suffered from a severe loss of muscle fibers and
profound signs of dystrophy in his tibialis muscle . The AON was well tolerated
and did not provoke serious adverse events in any of the patients. Our results
provide a strong basis for subsequent studies on systemic treatment of DMD
patients.
21 - Genetic treatments in muscular dystrophies
19 - 18 years experience with mechanical ventilation in patients with Duchenne muscular dystrophy
16 - (The 2nd Congress of Asian Society for Pediatric Research - 2007)
1) TREATMENT OF DUCHENNE MUSCULAR DYSTROPHY BY AMINOGLYCOSIDES
Lai PS, Lim PPD, Tay SKH, Low PS - Dept of Paediatrics, National University of Singapore
Background
Duchenne muscular dystrophy (DMD) is an X-linked disorder caused by absence of dystrophin protein. A novel genetic approach for therapy of DMD patients carrying nonsense mutations has been proposed using aminoglycoside treatment. Aminoglycosides are postulated to suppress nonsense mutations by translational readthrough of stop codons, thus resulting in protein expression. The aim of this study is to investigate the efficacy of four aminoglycosides (gentamicin, paromomycin, tobramycin and G418), and to develop cell-free and cell-based assays for measuring readthroughs.
Methods
The cell-free system involved in vitro transcription-translation reactions to assay the radioisotope-labeled protein produced after aminoglycoside treatment. The cell-based system was designed by cloning mutational constructs carrying readthrough cassettes of three types of stop codons, UAA, UAG and UGA. The constructs were transfected into HEK cell lines treated with aminoglycosides and assayed by FITC fluorescence.
Results
For the cell-free assays, optimal readthroughs of the UAG stop codon was observed ranging from 5-20% (2-10 mcg/ml gentamicin), 20-40% (20-400 mcg/ml paromomycin), 6-14% (80-1000 mcg/ml tobramycin) and 2-20% (2-1000 mcg/ml G418). For the cell-based assays, the readthroughs were lower, with maximum values of only up to 3%. The optimal concentrations for the aminoglycosides were : 0.75–1.0 mg/ml gentamicin, 1–2.5 mg/ml paromomycin, > 2.4 mg/ml tobramycin, 0.15- 2.4 mg/ml G418.
Conclusions
The nonsense mutation most susceptible to aminoglycoside suppression is UGA stop codon. All aminoglycosides resulted in some levels of readthroughs. Differences in clinically effective readthrough levels between the two assay systems highlight the need for caution when comparing results using different assays.
2) Purification of Satellite Cells From Mouse ES Cells
Hsi Chang1, Toshio Heike1, Momoko Yoshimoto2, Katsutsugu Umeda1, Tatsutoshi Nakahata1
1Kyoto University, Department of Pediatrics, 2Wells Center For Pediatric Research, Indianapolis, IN, USA
Duchene muscular dystrophy (DMD) is well known as one of the progressive muscle diseases which decline the quality of life (QOL). Although many researchers search for new possible and trial therapies, but there is no conclusive clinical therapy that could improve the patients’ QOL till nowadays. For the solution of these problems, myogenic cell transplantation has been reported as a new approach recently. Before the establishment of myogenic cell transplantation therapy, the identification and purification of muscle specific stem cells called satellite cells could be indispensable. So far, there have been many studies related to the isolation and characterization of satellite cells. However, the differentiation and isolation of satellite cells from mouse embryonic stem (ES) cells has not yet been reported and it is still hard to specifically induce myogenic cells from ES cells. Here, we report a newly established method that effectively induces a cell subpopulation with satellite cell properties from mouse ES cells. This is the first report of satellite cells induced from mouse ES cells and these findings could pave a path for therapeutic use of ES cells for muscular disorders.
3) Comprehensive mutation analysis of the dystrophin gene in Japanese Duchenne and Becker muscular dystrophies
Yasuhiro Takeshima, Kazuto Ishibashi, Atsushi Nishiyama, Zhujun Zhang, Tran Thi Hoai Thu, Yasuaki Habara, Mariko Yagi, Masafumi Matsuo - Kobe University Graduate School of Medicine, Department of Pediatrics
[Background] Duchenne and Becker muscular dystrophy (DMD/BMD) are caused by mutations in the dystrophin gene. Mutation analysis of the dystrophin gene is indispensable not only to provide proper clinical information but to apply the molecular therapies, which depend on the type of mutation in each case. However, the large size of dystrophin gene (3000kb and 79 exons) hampers the detection of small mutations including nonsense, frame shift, and splice site mutations. To clarify the responsible mutations of all cases, a comprehensive mutation analysis of the dystrophin gene employing not only genomic DNA but mRNA was performed in Japanese DMD/BMD. [Patients and Methods] A total of 431 cases from unrelated 355 Japanese families of DMD/BMD were recruited. Gross gene arrangements were detected by Southern blotting or PCR methods using genomic DNA as template, and small mutations were analyzed by RT-PCR or direct sequence method using genomic DNA or cDNA as template. [Results] Among 355 families, 221 (62%) and 23 (6%) had large deletion and duplication encompassing at least one exon, respectively. X-chromosome abnormality was disclosed in 3 families. In the 108 remaining families, 61 nonsense, 23 frame shift, and 22 splice site mutations were identified. It is noteworthy that 67% of them are novel mutations. In two cases, however, no mutation was detected.[Conclusions] In this study we succeeded to reveal the responsible gene mutations in 99.4% of the analyzed cases, and the present result disclosed the highest mutation detection rate in DMD/BMD.
15 - (Spine, 32(20), 2007, 2278-2283) Tranexamic Acid Diminishes Intraoperative Blood Loss and Transfusion in Spinal Fusions for Duchenne Muscular Dystrophy Scoliosis
Shapiro, Frederic ; Zurakowski, David P; Sethna, Navil F - USA
Study Design. Retrospective review of intraoperative blood loss and blood replacement.
Objective. We compared intraoperative blood loss and blood replacement during spinal fusion surgery for scoliosis in Duchenne muscular dystrophy (DMD) performed with and without the synthetic antifibrinolytic agent tranexamic acid (TXA).
Summary of Background Data. High levels of intraoperative blood loss are widely documented in DMD patients undergoing posterior spinal fusion for scoliosis. The effect of the antifibrinolytic agent tranexamic acid on decreasing the blood loss has not been studied in a large group of DMD patients.
Methods. All 56 DMD patients underwent posterior spinal fusion with the same technique using 2 rods and multiple sublaminar wires. TXA was not used in 36 patients and was used in 20. In the respective groups, the age at surgery (14 vs. 13.9 years), the preoperative deformity (45° vs. 51°), the mean number of levels fused (14.3 vs. 14.7), and the mean surgical times (446 minutes vs. 459 minutes) were similar. TXA dose was 100 mg/kg in solution over 15 minutes before incision followed by an infusion of 10 mg/kg per hour during surgery. Standardized measurements of intraoperative blood loss were used and calculated to compare total amount of blood loss in milliliters per patient and blood loss as a percentage in relation to estimated blood volume [estimated blood loss (EBL)/estimated blood volume (EBV) × 100]. The EBV was calculated to be 70 mL/kg (body weight).
Results. Mean blood loss with TXA was 1944 ± 789 mL (range, 760–4000 mL) and without TXA was 3382 ± 1795 mL (range, 600–9580 mL) (P < 0.001). Blood loss with TXA decreased by 42% compared with those not treated with TXA. Accounting for patient weight and estimated blood volume, mean % blood loss with and without TXA was 47% ± 28% versus 112% ± 67% (P < 0.001). This physiologic indicator shows that blood loss with TXA decreased by 58% compared with those patients not treated with TXA. TXA was also found to reduce blood loss after accounting for surgical time. No hypercoagulation or other complications from TXA therapy were obs-erved. The reduced blood loss in TXA-treated patients translated into decreased blood transfusions. Transfusion of homologous whole blood and packed red blood cells in the TXA group was decreased by 46% compared with the no TXA group (mean levels, 512 ± 470 mL vs. 955 ± 718 mL), and transfusion of autologous cell saver blood was decreased by 42% in the TXA group (mean levels, 419 ± 235 mL vs. 728 ± 416 mL).
Conclusion. TXA significantly reduces both intraoperative blood loss and the need for homologous transfusion of whole blood and packed red blood cells in DMD patients undergoing posterior spinal fusion for scoliosis.
15 - CTS-IPITA-IXA 2007 Joint Conference – Abstracts - Minneapolis – September 15–20, 2007
13 - 6th Monaco Round Table report, held on June 23rd, 2007
12 - Dystrophin levels as low as 30% are sufficient to avoid muscular dystrophy in the human
11 - (Am J Physiol Heart Circ Physiol 293: H1969-H1977, 2007) The role of reactive oxygen species in the hearts of dystrophin-deficient mdx mice
Duchenne muscular dystrophy (DMD) is caused by deficiency of the cytoskeletal protein dystrophin. Oxidative stress is thought to contribute to the skeletal muscle damage in DMD; however, little is known about the role of oxidative damage in the pathogenesis of the heart failure that occurs in DMD patients. The dystrophin-deficient (mdx) mouse is an animal model of DMD that also lacks dystrophin. The current study investigates the role of the antioxidant N-acetylcysteine (NAC) on mdx cardiomyocyte function, Ca2+ handling, and the cardiac inflammatory response. Treated mice received 1% NAC in their drinking water for 6 wk. NAC had no effect on wild-type (WT) mice. Immunohistochemistry experiments revealed that mdx mice had increased dihydroethidine (DHE) staining, an indicator of superoxide production; NAC-treatment reduced DHE staining in mdx hearts. NAC treatment attenuated abnormalities in mdx cardiomyocyte Ca2+ handling. Mdx cardiomyocytes had decreased fractional shortening and decreased Ca2+ sensitivity; NAC treatment returned mdx fractional shortening to WT values but did not affect the Ca2+ sensitivity. Immunohistochemistry experiments revealed that mdx hearts had increased levels of collagen type III and the macrophage-specific protein, CD68; NAC-treatment returned collagen type III and CD68 expression close to WT values. Finally, mdx hearts had increased NADPH oxidase activity, suggesting it could be a possible source of increased reactive oxygen species in mdx mice. This study is the first to demonstrate that oxidative damage may be involved in the pathogenesis of the heart failure that occurs in mdx mice. Therapies designed to reduce oxidative damage might be beneficial to DMD patients with heart failure.
11 - (Arch Neurol. 2007;64:1236-1241) Gene Therapy for Duchenne Muscular Dystrophy: Expectations and Challenges
Duchenne muscular dystrophy is a debilitating X-linked disease with limited treatment options. We examined the possibility of moving forward with gene therapy, an approach that demonstrates promise for treating Duchennemuscular dystrophy. Gene therapy is not limited to replacement of defective genes but also includes strategies using surrogate genes with alternative but effective means of improving cellular function or repairing gene mutations. The first viral-mediated gene transfer for any muscle disease was carried out at Columbus Children's Research Institute and Ohio State University for limb girdlemuscular dystrophy type 2D, and the first viral-mediated trial of gene transfer for Duchenne muscular dystrophy is under way at the same institutions. These studies, consisting of intramuscular injection of virus into a single muscle, are limited in scope and represent phase 1 clinical trials with safety as the primary end point. These initial clinical studies lay the foundation for future studies, providing important information about dosing, immunogenicity, and viral serotype in humans. This article highlights the challenges and potential pitfalls as the field advances this treatment modality to clinical reality.
Myogenic precursor cell (MPC) transplantation is a good strategy to introduce dystrophin expression in muscles of Duchenne muscular dystrophy (DMD) patients. Insulin-like growth factor (IGF-1) promotes MPC activities, such as survival, proliferation, migration and differentiation, which could enhance the success of their transplantation. Alternative splicing of the IGF-1 mRNA produces different muscle isoforms. The mechano growth factor (MGF) is an isoform, especially expressed after a mechanical stress. A 24 amino acids peptide corresponding to the C-terminal part of the MGF E domain (MGF-Ct24E peptide) was synthesized. This peptide had been shown to enhance the proliferation and delay the terminal differentiation of C2C12 myoblasts. The present study showed that the MGF-Ct24E peptide improved human MPC transplantation by modulating their proliferation and differentiation. Indeed, intramuscular or systemic delivery of this synthetic peptide significantly promoted engraftment of human MPCs in mice. In vitro experiments demonstrated that the MGF-Ct24E peptide enhanced MPC proliferation by a different mechanism than the binding to the IGF-1 receptor. Moreover, MGF-Ct24E peptide delayed human MPC differentiation while having no outcome on survival. Those combined effects are probably responsible for the enhanced transplantation success. Thus, the MGF-Ct24E peptide is an interesting agent to increase MPC transplantation success in DMD patients.
AUGUST
28 - Quadrupling Muscle Mass in Mice by Targeting TGF-ß Signaling Pathways
28 - (Neuromuscular Disorders, 2007) Workshop report: Behavior patterns in Duchenne muscular dystrophy: Report on the Parent Project Muscular Dystrophy behavior workshop 8–9 of December 2006, Philadelphia, USA LIBRARY
James Poysky - USA
Introduction
Eighteen participants representing clinicians, scientists, parents, industry, and public health agencies involved in the assessment and treatment of children with Duchenne muscular dystrophy (DMD) from Australia, Canada, the Netherlands, the UK and the USA met in Philadelphia on December 8–9, 2006 to attend a workshop addressing behavioral issues among children and young adults with DMD. The workshop, sponsored by Parent Project Muscular Dystrophy (PPMD–USA), was organized in response to parent and clinician concerns regarding behavior associated with DMD, and its subsequent impact on patient and family quality of life. For the purposes of clarification, in this report the term “behavior” is defined as readily observable external actions and responses, as well as internal affective/emotional states. The aims of the workshop were to (1) evaluate current findings and determine whether characteristic behaviors occur among boys and young men with DMD, (2) examine etiological factors contributing to behavior patterns in DMD, and (3) develop recommendations for research and intervention strategies.
DMD is a genetic disorder occurring in approximately one in 3300 live male births. Mutations in the dystrophin gene result in an absence of dystrophin or a non-functional dystrophin protein. The different types and locations of the mutation are highly variable across individuals, and it is unclear to what extent this has an impact on the phenotype. However, in all cases progressive muscle degeneration ensues, with morbidity typically occurring in the second or third decade of life. Muscle degeneration has understandably been the primary focus of research and treatment, and much less emphasis has been placed on behavioral functioning. There is, however, emerging evidence of central nervous system (CNS) involvement resulting in cognitive and neurobehavioral disorders. In addition, because of the debilitating and fatal nature of the disorder, significant psychosocial factors must also be considered. Behavioral functioning has a significant impact on quality of life and can also directly affect medical care, in particular as it is related to things like treatment adherence and behavioral response to medication. Research to date is limited, and studies have typically focused on one component of physiological, cognitive, or emotional functioning in DMD. The complex interactions between these various factors have not been clearly defined. As such, greater understanding of these interactions is necessary for appropriate patient care and treatment planning.
25 - (Neuromuscular Disorders, 2007) Case report: Myoglobinuria in boys with Duchenne muscular dystrophy on corticosteroid therapy
P. Garrood, M. Eagle, P.E. Jardine, K. Bushby and V. Straub - UK
Myoglobinuria is a recognised complication of Duchenne muscular dystrophy (DMD), but has only once been reported in ambulant boys on corticosteroid therapy [Dubowitz V, Kinali M, Main M, Mercuri E, Muntoni F. Remission of clinical signs in early Duchenne muscular dystrophy on intermittent low-dosage prednisolone therapy. Eur J Paediatr Neurol 2002;6(3):153–9.]. We present three prednisolone-treated boys with myoglobinuria and in two cases this was recurrent. All three showed improved motor performance in response to the introduction of corticosteroids. The greater activity of steroid-treated individuals may place their dystrophin-deficient muscles under greater mechanical stress, predisposing to further muscle fibre damage and consequent myoglobinuria. Families and physicians need to have an increased awareness of this possibility and of the appropriate management of myoglobinuria.
25 - Videofluorographic assessment of swallowing function in patients with Duchenne muscular dystrophy
25 - Utrophin up-regulation by an artificial transcription factor in transgenic mice
22 - (Pediatric Anesthesia,2007) Duchenne muscular dystrophy: an old anesthesia problem revisited LIBRARY
Jason Hayes, Francis Veyckemans, Bruno Bissonnette - Canada e Belgium
Patients with Duchenne and Becker muscular dystrophy suffer from a progressive deterioration in muscle secondary to a defect in the dystrophin gene. As such, they are susceptible to perioperative respiratory, cardiac and other complications, such as rhabdomyolysis. Inhalational anesthetic agents have been implicated as a cause of acute rhabdomyolysis that can resemble malignant hyperthermia (MH). This article reviews perioperative ‘MH-like’ reactions reported in muscular dystrophy patients and groups them into three categories according to clinical presentation. The etiology and underlying pathophysiological process responsible for these reactions is discussed and recommendations are proposed for the safe anesthetic management of these patients.
13 - (Brain & Development 29:596-500,2007) Cardiac troponin I for accurate evaluation of cardiac status in myopathic patients
Tsuyoshi Matsumura , Toshio Saito, Harutoshi Fujimura, Susumu Shinno - Japan
Background: Brain natriuretic peptide (BNP) and left ventricular ejection fraction (LVEF) are standard indexes for cardiac function. However, they can not reveal myocardial damage directly and they often remain normal even in advanced cardiomyopathy in immobilized patients. Myocardial markers such as MB type of creatine kinase (CK-MB), heart-type fatty acid binding protein (HFABP) and cardiac troponins are expected to evaluate active myocardial degeneration. However, their availabilities in these patients have not been examined yet. Methods: Participants were 129 patients with dystrophinopathies; 100 Duchenne muscular dystrophy (DMD), 25 Becker muscular dystrophy (BMD) and 4 DMD/BMD carriers. Various serological cardiac indexes, including CK-MB, H-FABP, cardiac troponin I (cTnI), BNP and LVEF were measured and statistical analysis was done. Results: CK-MB and HFABP was highly associated with creatine kinase (CK). On the contrary, cTnI, BNP and LVEF were independent from CK. In DMD, relatively high cTnI values were observed in patients with motor ability of rowing wheelchair and in their second decade. BNP and LVEF was strongly correlated. However, cTnI was independent from LVEF and only weak correlation could be detected between cTnI and BNP. Conclusion: cTnI had been proven to be expressed in myocardium exclusively. Our results also certified that cTnI can assess cardiac degeneration independently from skeletal muscle degeneration and is practical index even in myopathic patients. Our findings also suggested that cardiac degeneration was preceded to functional impairment in many cases. It indicated that cTnI enable us to detect early stage of cardiac degeneration and initiate intervention at proper stage.
11- 2007 Annual Conference Powerpoint Presentations
JULY
31 - (Growth Hormone & IGF Research, 2007) Muscular dystrophy-related quantitative and chemical changes in adenohypophysis GH-cells in golden retrievers
A.R. de Lima , J.R. Nyengaard , A.A.L. Jorge , J.C.C. Balieiro , C. Peixoto , E.T. Fioretto , C.E. Ambrosio f, M.A. Miglino , M. Zatz , A.A.C.M. Ribeiro - Brazil
Duchenne muscular dystrophy (DMD) is a recessive X-linked lethal condition which affects a boy in every 3300 births. It is caused by the absence of dystrophin, a protein occurring especially within the musculoskeletal system and in neurons in specific regions of the central nervous system (CNS). Growth hormone (GH) inhibition is believed to decrease the severity of DMD and could perhaps be used in its treatment. However, the underlying pathological mechanism is not known. The golden retriever muscular dystrophy dog (GRMD) represents an animal model in the study of DMD. In this paper we investigated the morphological aspects of the adenohypophysis as well as the total number and size of GH-granulated cells using design-based stereological methods in a limited number of dystrophic and healthy golden retrievers. GH-cells were larger (32.4%) in dystrophic dogs than in healthy animals (p = 0.01) and they occupied a larger portion (62.5%) of the adenohypophysis volume (p = 0.01) without changes in either adenohypophysis volume (p = 0.893) or total number of GH-granulated cells (p = 0.869). With regard to ultrastructure, granulated cells possessed double-layer electron-dense granules which were evenly distributed in the cytosol. Furthermore, these granules in dystrophic animals occupied a larger proportion of GH-granulated cell volume (66.9%; p = 0.008) as well as of all GH-cells in the whole pars distalis of adenohypophysis (77.3%; p = 0.035), albeit IGF-1 serum concentration was lower in severe cases. This suggests difficulties in the GH secretion that might possibly be associated to dystrophin absence. In contrast to earlier reports, our data suggest that a lower IGF-1 concentration may be more related to a severe, as opposed to a benign, clinical form of muscular dystrophy.
31 - (International Journal of Cardiology,2007) Letter to the Editor: Holter electrocardiogram should be systematic in Duchenne muscular dystrophy
A. Fayssoil - France
Duchenne muscular dystrophy (DMD) is due to mutations in the dystrophin gene on chromosome Xp21.1. Dystrophin is a sarcolemmal protein which binds actin to extra cellular matrix. A deficiency in dystrophin leads to breakdown of muscle membrane and muscular alteration
[1]. The disease is clinically characterized by proximal weakness and wasting, leading to loss of ambulation by 12 years of age. The heart is affected to various degrees, depending on the stage of the disease. The most common cardiac abnormality in DMD is dilated cardiomyopathy. Conduction system disease and tachyarrhythmias may occur in some patients. Death occurs in early adulthood secondary to respiratory or cardiac failure [2], even if recent advances in the use of inspiratory and expiratory muscle aids have greatly reduced the risk of pulmonary morbidity and mortality rates [3].The typical initial manifestation of cardiac involvement in DMD is sinus tachycardia [1]. Cardiac involvement is due to progressive replacement of the cardiomyocytes and the Purkinje system by connective tissue or fat [1]. Impairment of cardiac autonomic function in patients with DMD is well known. Kirchmann found sinus tachycardia in 26% and reduced heart rate variability in 51% [4]. In human cardiac purkinje fibers, immunocytochemical staining showed that dystrophin was localised to the membrane surface of the purkinjer fiber [5]. The cardiac rhythm abnormalities play a significant role in morbidity and mortality in this disease [6]. Supraventricular ectopy or couplets are reported to indicate increasing severity of arrhythmia with the progression of disease [7]. 33% of DMD patients had ventricular premature beats in a study including 45 Duchenne muscular dystrophy patients without congestive heart failure and followed up for 3 years [8]. Patients who died suddenly were more likely to have had documented complex ventricular arrhythmias (6 of 9; 66%) [8].Shortened PQ interval and prolonged QT interval are also reported. In a controlled series including 328 DMD patients, 62% had conduction abnormalities like shortened PQ interval and prolonged QT interval by age 10 years [9]. Conduction system disease was also reported. Indeed, complete atrioventricular block was reported in the literature in 1997 [10]. Holter electrocardiogram should be systematic periodically in the following of the DMD patients because of possible occurring conduction system disease and tachyarrhythmias. According to American paediatrics recommendations, periodic Holter monitoring should be considered for patients with demonstrated cardiac dysfunction [2].References
[1] Finsterer J, Stollberger C. The heart in human dystrophinopathies cardiology. 2003; 99: 1-19.
[2] American Academy of Pediatrics Section on Cardiology and Cardiac Surgery. Cardiovascular health supervision for individuals affected by Duchenne or Becker muscular dystrophy. Pediatrics Dec 2005;116(6):1569–73.
[3] Bach JR, Ishikawa Y, Kim H. Prevention of pulmonary morbidity for patients with Duchenne muscular dystrophy. Chest 1997;112:1024–8. [4] Kirchmann C, Kececioglu D, korinthenberg R, Dittrich S. Echocardiographic and electrographic finding of cardiomyopathy in Duchenne and Becker-Kiener muscular dystrophies. Pedriatr Cardiol janv-feb 2005;26(1):66–72.
[5] Bies RD, Friedman D, Roberts R, Perryman MB, Caskey CT. Expression and localization of the dystrophin in human cardiac purkinje fibers. Circulation 1992;86(1):147–53.
[6] Corrado G, Lissoni A, Beretta S, et al. Prognostic value of electrocardiograms, ventricular late potentials, ventricular arrhythmias, and left ventricular systolic dysfunction in patients with Duchenne muscular dystrophy. Am J Cardiol 2002;89:838–41.
[7] Yanagisawa A, Miyagawa M, Yotsukura M, et al. The prevalence and prognostic significance of arrhythmias in Duchenne muscular dystrophy. Am Heart J 1992;124:1244–50
[8] Chenard AA, Becane HM, Tertrain F, de Kermadec JM, Weiss YA. Ventricular arrhythmia in Duchenne muscular dystrophy: prevalence, significance and prognosis. Neuromuscul Disord May 1993;3(3):201–6. [9] Nigro G, Comi L, Politan L, Brain RJ. The incidence and evolution of cardiomyopathy in Duchenne muscular dystrophy. Int J Cardiol 1990;26:271–7.
[10] Takano N, Honke K, Hasui M, Ohno I, Takemura H. A case of pacemaker implantation for complete atrioventrivular block associated with Duchenne muscular dystrophy. No to Hattatsu Nov 1997;29 (6):476–80.
Mechanism of action of ACE inhibitors and Angiotensin II blockers
27 - Episodic hypoxia exacerbates respiratory muscle dysfunction in DMD(mdx) mice
K. Bushby , R. Griggs , on behalf of the MSG/ENMC FOR DMD trial study group
Eighteen participants from 6 countries met in Naarden from 22nd–24th October 2006 to review the proposed plan for an international trial to find the optimal steroid regimes in Duchenne Muscular Dystrophy (DMD)(trial acronym FOR DMD). At the 124th ENMC international workshop in April 2004 the need for a trial of corticosteroid dosage regimes in DMD was highlighted by clinicians in the field as well as patient organisations, reflecting a high level of dissatisfaction with the inconsistency of use of corticosteroid and the profusion of different steroid regimes in use in clinics across the world. The conclusion of that workshop was that a trial of different corticosteroid regimes was indicated, and subsequently an international steering committee was established. This committee prepared a protocol for an international trial of steroid dosage regimes in DMD which received funding from the NIH for a planning grant to develop a full trial grant application. The aim of the current workshop was to bring together members of the trial writing group with representatives of investigators from the different countries taking part in the trial to allow a full review of the protocol and prepare the final version of the protocol and manual of operations to be submitted as a full grant to NIH in early 2007.
Table included in this article:
25 - Arrhythmia follow-up of children and adolescents with neuromuscular diseases
24 - (American Heart Journal, 2007) Perindopril preventive treatment on mortality in Duchenne muscular dystrophy: 10 years' follow-up
Denis Duboc; Christophe Meune, Bertrand Pierre, Karim Wahbi, Bruno Eymard, Annick Toutain, Carole Berard, Guy Vaksmann, Simon Weber, Henry-Marc Becane - France
Background: Duchenne muscular dystrophy (DMD), an X-linked disorder due to lack of dystrophin, is associated with muscle weakness and myocardial dysfunction. Although preliminary data support the efficacy of angiotensin-converting enzyme inhibitors on left ventricular (LV) function, our aim was to examine the long-term impact of a preventive treatment with perindopril on mortality in children with DMD.
Methods: Patients with DMD between the ages of 9.5 and 13 years presenting with normal LV ejection fraction were included in this prospective study. They were randomly assigned for 3 years to perindopril, 2 to 4 mg (group 1), or placebo (group 2) in a double-blind protocol, followed by open-label treatment with perindopril for up to 10 years. Survival rate at 10 years in each group is reported.
Results: There were 28 patients assigned to group 1 and 29 to group 2. Baseline characteristics were similar in both groups. At the end of the 10 years' follow-up period, survival status was available for all included patients: 26 (92.9%) of 28 patients in group 1 were alive at 10 years versus 19 (65.5%) of 29 in group 2 (P = .02). Kaplan-Meier cumulative survival was significantly lower in group 2 than in group 1 (P = .013).
Conclusion: Early initiation of treatment with perindopril is associated with a lower mortality in patients with DMD with normal LV ejection fraction at study entry.
14 - Low bone mineral density and decreased bone turnover in Duchenne muscular dystrophy
12 - New Target For Muscular Dystrophy Drug Therapy Found
ERF Silences Extrasynaptic Utrophin by N-Box-mediated Repression in Skeletal Muscle
7 - Antisense-induced exon skipping for duplications in Duchenne muscular dystrophy
Myostatin (Mst) is a negative regulator of skeletal muscle in humans and animals. It is moderately expressed in the heart of sheep and cattle, increasing considerably after infarction. Genetic blockade of Mst expression increases cardiomyocyte growth. We determined whether Mst overexpression in the heart of transgenic mice reduces left ventricular size and function, and inhibits in vitro cardiomyocyte proliferation. Young transgenic mice overexpressing Mst in the heart (Mst transgenic mice (TG) under a muscle creatine kinase (MCK) promoter active in cardiac and skeletal muscle, and Mst knockout (Mst (–/–)) mice were used. Xiscan angiography revealed that the left ventricular ejection fraction did not differ between the Mst TG and the Mst (–/–) mice, when compared with their respective wild-type strains, despite the decrease in whole heart and left ventricular size in Mst TG mice, and their increase in Mst (–/–) animals. The expected changes in cardiac Mst were measured by RT-PCR and western blot. Mst and its receptor (ActRIIb) were detected by RT-PCR in rat H9c2 cardiomyocytes. Transfection of H9c2 with plasmids expressing Mst under muscle-specific creatine kinase promoter, or cytomegalovirus promoter, enhanced p21 and reduced cdk2 expression, when assessed by western blot. A decrease in cell number occurred by incubation with recombinant Mst (formazan assay), without affecting apoptosis or cardiomyocyte size. Anti-Mst antibody increased cardiomyocyte replication, whereas transfection with the Mst-expressing plasmids inhibited it. In conclusion, Mst does not affect cardiac systolic function in mice overexpressing or lacking the active protein, but it reduces cardiac mass and cardiomyocyte proliferation.
JUNE
30 - New Genetic Test Developed At Emory Advances Detection And Diagnosis Of Muscular Dystrophy
21 - Morpholino Oligomer-Mediated Exon Skipping Averts the Onset of Dystrophic Pathology in the mdx Mouse
18 - (Biology of Blood and Marrow Transplantation, 2007) Hematopoietic Cell Transplantation Directly into Dystrophic Muscle Fails to Reconstitute Satellite Cells and Myofibers
Christian S. Kuhr, Marilena Lupu, Rainer Storb - USA
We sought to determine whether wild-type hematopoietic cell transplantation directly into muscle could restore dystrophin expression in a relevant preclinical canine model of Duchenne muscular dystrophy. In recipients rendered tolerant to their dog leukocyte antigen-matched unaffected littermates through hematopoietic stem cell transplantation, intramuscular injection of donor marrow cells produced no evidence of dystrophin expression, and clonal analysis of satellite cells failed to reveal any donor contribution.
16 - The role of reactive oxygen species in the hearts of dystrophin-deficient (mdx) mice
16 - Chronic respiratory failure in patients with neuromuscular diseases: diagnosis and treatment LIBRARY
9 - Potential of oligonucleotide-mediated exon-skipping therapy for Duchenne muscular dystrophy
8 - Living with muscular dystrophy: health related quality of life consequences for children and adults
7 - (Free Radical Biology and Medicine; 43(1):145-54) Low intensity training decreases markers of oxidative stress in skeletal muscle of mdx mice
Jan J. Kaczor, , Julie E. Hall, Eric Payne and Mark A. Tarnopolsky - Canada
Reactive oxygen species may contribute to the pathogenesis
of muscular dystrophy. High intensity exercise clearly
induces muscle damage in mdx mice; however, the effects of low intensity
exercise training (LIT) on mdx muscle are less clear. We examined the
effect of LIT on markers of oxidative stress (malondialdehyde and protein
carbonyls), antioxidant (superoxide dismutase, catalase, and glutathione
peroxidase), and mitochondrial (2-oxoglutarate dehydrogenase and cytochrome
oxidase) enzymes in skeletal muscle of mdx and wild-type mice. Mdx
and wild-type mice were allocated to LIT and sedentary groups. Malondialdehyde
levels were higher in white muscle from sedentary mdx as compared to both
sedentary and LIT wild-type mice (P < 0.001). Protein carbonyl content
was higher in white and red muscle of mdx versus wild-type mice (P < 0.05).
LIT was associated with lower levels of malondialdehyde and protein carbonyls in
white muscle of mdx mice (decreased 38 and 44%, P < 0.001 and P < 0.01,
respectively). Antioxidant and mitochondrial enzyme activities were higher in
white muscle of mdx than in wild-type mice (P < 0.05). LIT in
mdx mice induced physiological adaptation resulting in lower levels of
markers of oxidative stress that were not different than those from wild type.
These results are of relevance for therapeutic exercise in patients with
dystrophinopathy where exercise prescription remains controversial.
7 -
Herpes
Simplex Virus VP22 Enhances Adenovirus-Mediated Microdystrophin Gene Transfer to
Skeletal Muscles in Dystrophin-Deficient (mdx) Mice
5 - Effects of a CRF2R agonist and exercise on mdx and wildtype skeletal muscle
2 - The role of corticosteroids in muscular dystrophy: A critical appraisal
MAY
25 - Passive in vivo elastography from skeletal muscle noise
23 - (Am J Physiol Regul Integr Comp Physiol, 2007 ) Reversal by relaxin of altered ileal spontaneous contractions in dystrophic (mdx) mice through a nitric oxide-mediated mechanism
Altered nitric oxide (NO) production/release is involved in gastrointestinal motor disorders occurring in dystrophic (mdx) mice. Since the hormone relaxin (RLX) can up-regulate NO biosynthesis, its effects on spontaneous motility and NO synthase (NOS) expression in the ileum of dystrophic (mdx) mice were investigated. Mechanical responses of ileal preparations were recorded in vitro via force-displacement transducers. Evaluation of the expression of NOS isoforms was performed by immunohistochemistry and Western blot. Normal and mdx mice were distributed into three groups: untreated, RLX-pretreated, vehicle-pretreated. Ileal preparations from the untreated animals showed spontaneous muscular contractions whose amplitude was significantly higher in mdx than in normal mice. Addition of RLX, alone or together with L-arginine, to the bath medium depressed the amplitude of the contractions in the mdx mice, thus re-establishing a motility pattern typical of the normal mice. The NOS inhibitor L-NNA or the guanylate cyclase inhibitor ODQ reversed the effects of RLX. In RLX-pretreated mdx mice, the amplitude of spontaneous motility was reduced, thus resembling that of the normal mice, and NOS II expression in the muscle coat was increased in respect to the vehicle-pretreated mdx animals. These results indicate that RLX can reverse the altered ileal motility of mdx mice to a normal pattern, likely by up-regulating NOS II expression and NO biosynthesis in the ileal smooth muscle.
18 - (Acta Histochemica, 2007) Fibre-related nitric oxide synthase (NOS) in Duchenne muscular dystrophy
K. Punkt, S. Schering, M. Fritzsche, G. Asmussen, E.A. Minin, V.E. Samoilova, F.-U. Mu¨ller, W. Schmitz, M. Hasselblatt, W. Paulus, U. Muller-Werdan, J. Slezak, G. Koehler, W. Boecker, I.B. Buchwalow - Germany
Summary
Nitric oxide (NO) mediates fundamental physiological actions on skeletal muscle. The loss of NO synthase (NOS) from the sarcolemma was assumed to be associated with development of Duchenne muscular dystrophy (DMD). We have, however, recently reported that, in contrast to the commonly accepted view, NOS expression in DMD myofibres is up-regulated. This poses the question of the fibre type-specific NOS expression in DMD muscles and how the NOS expression is related to the regeneration or degeneration status. To address this issue, we examined localization of NOS isoforms I, II and III in skeletal muscles of DMD patients employing immunohistochemical labelling with tyramide signal amplification complemented with enzyme histochemistry. We found that NOS immunolabelling as well as metabolic enzyme activity in DMD muscles were heterogeneously distributed along the fibre length of DMD muscle fibres revealing regenerating and degenerate (hypercontracted) fibres as well as normal segments. Like in normal muscles, positive NOS immunoreactivity was found to be associated with fast-oxidative glycolytic (FOG) phenotype. The regeneration status of NOS-positive segments was deduced from the presence of neonatal and developmental myosin heavy chains. High NOS expression in regenerating DMD muscle fibres can be well reconciled with reports about the protective role of endogenous NO in inflammatory diseases and in muscle repair.
16 - Red-green color vision impairment in duchenne muscular dystrophy
12 - (Invest Ophthalmol Vis Sci 2007;48: E-Abstract 5277) Extraocular Muscles Buffer Calcium Better Than Limb Muscle: Implications for Preferential Sparing in Duchenne's Muscular Dystrophy
Abstract
Purpose:Duchenne's Muscular Dystrophy (DMD) is the most common fatal, genetic disease in males and associated with widespread muscle wasting. Extraocular muscles (EOMs) are spared in DMD, although the reasons for this preferential sparing remain elusive. Dysregulation of calcium homeostasis has been suggested to contribute to muscle damage in DMD. Based on morphological observations, pharmacological tests and expression profiling, we propose that EOM sparing is facilitated by differential calcium buffering properties of EOM compared to limb (tibialis anterior-TA) muscle. We investigated the role of candidate proteins as well as compared calcium handling properties of cultured myotubes from EOM and TA.
Methods:The sarcoplasmic reticulum (SR) content of EOM and TA muscle was calculated from electron microscopy (EM) micrographs. Quantitative PCR was used to compare mRNA expression levels. Primary myoblasts from EOM and TA from 5-10-day old Sprague Dawley rats were cultured and fused in vitro, using the Rando & Blau (1994) protocol. Intracellular calcium buffering was determined using Fura-2 loaded myotubes after challenging the cells with ionomycin and 300nM extracellular Ca2+.
Results:EM morphometry showed a 1.7 fold larger SR area in pale global EOM fibers than TA. The expression of mRNAs for several calcium and SR related proteins including SERCA1, Phospholamban and S100a1 was higher in EOM than TA. Cultured EOM myotubes showed 2.3 fold larger calcium peaks in response to sudden increases in calcium. However, the signal decayed 1.7 fold faster in EOM than in TA, implying a more efficient buffering from excessive calcium levels in these muscles.
Conclusions:Larger SR and differential expression of calcium buffering proteins may contribute to the improved ability of EOM to handle elevated intracellular calcium levels. This in turn would be predicted to protect EOM from the calcium-mediated damage noted in DMD limb muscles.
12 - (Invest Ophthalmol Vis Sci 2007;48: E-Abstract 3772) Upregulation of Retinal Dystrophin Dp260 Transcription by Methylprednisolone is Mediated by Glucocorticoid Receptor
Abstract
Purpose:The Duchenne muscular dystrophy (DMD) gene encodes several isoforms of dystrophin via either internal or external promoters. Internal promoters are located within introns, and their regulatory mechanisms are not well described. Dp260, also known as retinal dystrophin, is a 260kDa isoform of dystrophin that is expressed in the outer-plexiform layer of the retina and is associated with normal retinal electrophysiology. We sought to characterize the Dp260 internal promoter within DMD intron 29 and to identify elements that regulate its expression.
Methods:The human Dp260 promoter region was cloned, sequenced, and analysed for potential regulatory elements. A luciferase reporter-promoter construct was transfected into cell lines and exposed to methylprednisolone and the glucocorticoid receptor (GR) antagonist RU486. Transcriptional activity was measured. Potential transcription factors were characterized by EMSA.
Results:Two glucocorticoid response elements were identified by sequence analysis. Transcription was upregulated in Y79 (retinoblastoma), C2 (myoblast), and HeLa cells, but not in the GR-deficient COS-7 cell line. Transcription was blocked by RU486. EMSA showed a shift in the presence of a GRE DNA sequence and a supershift with anti-GRbeta.
Conclusions:The transcription of Dp260 can be manipulated by the use of steroid drugs and the effect is mediated by the glucocorticoid receptor binding to glucocorticoid response elements. This is of particular interest in the light of current steroid clinical treatment trials for DMD. An ability to manipulate Dp260 expression by pharmacogenetic means may have implications for the treatment of DMD and inherited disorders of night vision.
9 - (Neuromuscular Disorders, 2007) Managing Duchenne muscular dystrophy – The additive effect of spinal surgery and home nocturnal ventilation in improving survival
Michelle Eagle, John Bourke, Robert Bullock, Mike Gibson, Volker Straub and Kate Bushby
- United KingdomAbstract
Objectives: To determine the long term survival in patients with Duchenne muscular dystrophy (DMD) following spinal surgery and nocturnal ventilation.
Study design: A retrospective review of 100 consecutive patients born between 1970 and 1990 was conducted.
Results: Forty-seven patients had surgical spinal fusion, 27 were subsequently ventilated. Fourteen patients received ventilation only. Thirty-nine patients received neither intervention. The age at which ventilation was required correlated with the age at which ambulation was lost. Those who walked for longer were less likely to require spinal surgery. Mean vital capacity dropped from 1.4 to 1.13 L 1 year post-operatively. Patients having both spinal surgery and ventilation had a median survival of 30 years whereas those who were only ventilated survived to 22.2 years.
Conclusion: Nocturnal ventilation improves survival in DMD. Spinal surgery does not increase forced vital capacity but in combination with nocturnal ventilation further improves median survival to 30 years.
7 - (Neurology,
2007.68(19):1607-13) Orthopedic outcomes of long-term daily
corticosteroid treatment in Duchenne muscular dystrophy
King, W M. ; Ruttencutter, R ; Nagaraja, H N. ; Matkovic, V ; Landoll, J ; Hoyle, C ; Mendell, J R. ; Kissel, J T. - USA
Objective: To document the effects of long-term daily corticosteroid treatment on a variety of orthopedic outcomes in boys with Duchenne muscular dystrophy.
Methods: We reviewed the charts of 159 boys with genetically confirmed dystrophinopathies followed at the Ohio State University Muscular Dystrophy Clinic between 2000 and 2003. Charts were reviewed for ambulation status, type and duration of steroid treatment (if any), and orthopedic complications including presence and location of long bone fractures, vertebral compression fractures, and the presence and degree of scoliosis.
Results: The cohort consisted of 143 boys (16 boys with Becker dystrophy were excluded); 75 had been treated with steroids for at least 1 year, whereas 68 boys had never been treated or had received only a brief submaximal dose. The mean duration of daily steroid treatment was 8.04 years. Treated boys ambulated independently 3.3 years longer than the untreated group (p < 0.0001) and had a lower prevalence of scoliosis than the untreated group (31 vs 91%; p < 0.0001). The average scoliotic curve was also milder in the treated group (11.6°) compared with the untreated group (33.2°; p < 0.0001). Vertebral compression fractures occurred in 32% of the treated group, whereas no vertebral fractures were discovered in the steroid naive group (p = 0.0012). Long bone fractures were 2.6 times greater in steroid-treated patients.
Conclusions: Although boys with Duchenne muscular dystrophy on long-term corticosteroid treatment have a significantly decreased risk of scoliosis and an extension of more than 3 years' independent ambulation, they are at increased risk of vertebral and lower limb fractures compared with untreated boys.
7 - CureDuchenne Funds Dr. Giulio Cossu’s Adult Stem Cell Study
5 - Respiration and Muscular Dystrophy - 6 new researchs
2 - Gene Therapy in Muscular Dystrophy - 39 new unpublished researchs
1 - (Nature Medicine, 2007) SIK1 is a class II HDAC kinase that promotes survival of skeletal myocytes
During physical exercise, increases in motor neuron activity stimulate the expression of muscle-specific genes through the myocyte enhancer factor 2 (MEF2) family of transcription factors. Elevations in intracellular calcium increase MEF2 activity via the phosphorylation-dependent inactivation of class II histone deacetylases (HDACs). In studies to determine the role of the cAMP responsive element binding protein (CREB) in skeletal muscle, we found that mice expressing a dominant-negative CREB transgene (M-ACREB mice) exhibited a dystrophic phenotype along with reduced MEF2 activity. Class II HDAC phosphorylation was decreased in M-ACREB myofibers due to a reduction in amounts of Snf1lk (encoding salt inducible kinase, SIK1), a CREB target gene that functions as a class II HDAC kinase. Inhibiting class II HDAC activity either by viral expression of Snf1lk or by the administration of a small molecule antagonist improved the dystrophic phenotype in M-ACREB mice, pointing to an important role for the SIK1-HDAC pathway in regulating muscle function.
Inhibiting class II HDAC activity by the administration of a small molecule antagonist = trichostatin A
APRIL
28 - PAPERS THAT WILL BE PRESENTED IN 59TH ANNUAL MEETING OF AMERICAN ACADEMY OF NEUROLOGY, BOSTON, 2007
28 - (Bone, 2007) EFFECTS OF BISPHOSPHONATES ON BONE MASS AND THE INCIDENCE OF FRACTURES IN STEROID-TREATED PATIENTS WITH DUCHENNE MUSCULAR DYSTROPHY.
R. R. Scott, A. Huynh, N. Alos, Y. Brousseau, G. Chabot - Canada
Introduction: Duchenne muscular dystrophy (DMD) is the most common muscular dystrophy of childhood. Affected males present with muscle weakness and abnormal gait. Death usually occurs in the second or third decade of life. Osteoporosis and pathologic fractures are known consequences of DMD and affected boys have a low bone mass that declines progressively with loss of ambulation. Previous studies have shown that treatment with glucocorticoids preserves muscle strength and prolongs ambulation. Prednisone and deflazacort are equally effective but both contribute to bone loss.
Methods: A retrospective chart review identified 43 patients with DMD treated with deflazacort (0.9 mg/kg/day) between 1993 and 2005. Starting in 1998, bone mass was measured annually by DEXA using the Hologic QDR 4500. Bisphosphonate treatment was initiated in patients with fractures or significant pain. Twenty-eight patients were treated with alendronate (35 to 70 mg per week). Four patients received IV pamidronate (9 mg/kg/year). Testosterone therapy was given to patients without spontaneous puberty by age15.
Results: Patients were treated with deflazacort alone for an average of 5.6 years and bisphosphonates for an average of 4.0 years. Nineteen patients treated with alendronate and four patients treated with pamidronate had sufficient available data to assess progression in bone mass. The annual change in BMC, BMD, volumetric BMD, and BMD Z score was significantly higher during treatment with bisphosphonates (p<0.005). The difference between treatment with alendronate and pamidronate was only significant for BMD Z score (p=0.02) although the number of patients treated with pamidronate was small. During the study period, 84% of all patients sustained a fracture, of which more than half were vertebral compression fractures. During treatment with deflazacort alone, 68% of patients sustained fractures. The peak incidence of fractures was during the 6th year of treatment. After initiation of bisphosphonates, 37% of treated patients sustained fractures. The peak incidence of fractures was during the first 2 years of bisphosphonate therapy.
Conclusions: Patients with DMD treated with deflazacort lose bone mass during treatment and the prevalence of fractures is very high. Bisphosphonate therapy improves their annual gain in bone mass and reduces their fracture risk.
28 - Effect of specific respiratory muscle training in mdx mouse on calcium homeostasis and mitochondrial respiration
J Gayraud, A Lacampagne, D Mornet, J Mercier, M Ramonatxo, S Matecki - France
Introduction: The cellular mechanisms resulting from the low exercice capacity of respiratory muscles in children with Duchenne muscular dystrophy (DMD) are poorly known. The aim of this study was thus to investigate the functional effect induced by a specific respiratory muscle training in mdx mouse diaphragm. Methods: Respiratory muscle training, was performed by hypercapnic stimulation of ventilation (8% CO2), during 6 weeks/30 min per day, in 10 mdx mice (10 mdx mice were also used as control).
Results: Diaphragmatic force frequency relationships showed that this specific respiratory training improves the tetanic force production of the diaphragm. Maximal mitochondrial respiration was increased in the training group (14.3 ± 0.4 vs. 9.5 ± 0.3 lmol/mn/mg) with no difference between the groups in diaphragm muscular fiber type and in citrate synthase activity. In additon, ryanodine receptor (RyR) activity was extrapolated from the analysis of spontaneous Ca2+-sparks measured on individual diaphragm fibers using laser scanning confocal microscopy. It showed decrease of Ca2+-spark rise-time in training group (4.66 ± 0.11 vs. 5.59 ± 0.12 ms) without any difference in Ca2+-spark amplitude, which could be interpreted as a better synchronisation of RyR opening involved in spark generation.
Conclusion: In conclusion, functional beneficial effect of low level of exercise on mdx diaphragm could be partially explained by an improvement of mitochondrial function and RyR gating properties which could attenuate the basal rise of cytoplasmic Ca2+.
28 - (Science in Sports & Exercise. 39(5):816-820, May 2007) Creatine Monohydrate Increases Bone Mineral Density in Young Sprague-Dawley Rats.
ANTOLIC, ANAMARIA ; ROY, BRIAN D. ; TARNOPOLSKY, MARK A. ; ZERNICKE, RONALD F. ; WOHL, GREGORY R. ; SHAUGHNESSY, STEPHEN G. ; BOURGEOIS, JACQUELINE M. - Canada
Introduction: Creatine kinase, found in osteoblasts,
is an enzyme that is upregulated in response to interventions that enhance bone
mass accretion. Creatine monohydrate supplementation can increase fat-free mass
in young healthy men and women and can reduce markers of bone breakdown in boys
with Duchenne muscular
dystrophy.
Purpose: The objective of this study was to determine the influence of
supplementation with creatine monohydrate on bone structure and function in
growing rats, to establish a therapeutic model.
Materials and Methods: Creatine monohydrate (2% w.w.) (CR; N = 16) or
standard rat chow (CON; N = 16) was fed to Sprague-Dawley rats beginning at 5 wk
of age, for 8 wk. Bone mineral density (BMD) and content (BMC) were assessed
using dual-energy x-ray absorptiometry at the beginning and end of the protocol.
The rats were sacrificed, and one femur was removed for the determination of
mechanical properties.
Results: The CR-treated rats showed greater lumbar BMD and femoral
bending load at failure compared with the CON rats (P < 0.05).
Conclusions: Together, these data suggest that creatine monohydrate
potentially has a beneficial influence on bone function and structure; further
investigation is warranted into its effect on bone functional properties and its
effects in disorders associated with bone loss.
24 - (European Journal of Paediatric Neurology, 2007) CINRG pilot trial of oxatomide in steroid-naıve Duchenne muscular dystrophy
Gunnar M. Buyse,, Nathalie Goemans, Erik Henricson, Alejandro Jara, Marleen van den Hauwe, Robert Leshner, Julaine M. Florence, Jill E. Mayhew, Diana M. Escolar - Belgium and USA
The authors report a pilot open-label two-center therapeutic trial of oxatomide in 14 steroid-naive DMD boys aged 5–10 years. Comparison of linear evolutions between 3 months medication-free lead-in periods and 6 months treatment periods showed no significant differences in quantitative (QMT) and manual (MMT) measurements of muscle strength and timed functional tests. A modest mitigation of strength deterioration over time cannot be excluded.
23 - (NATURE, 2007) PTC124 targets genetic disorders caused by nonsense mutations
21 - Rehabilitation interventions for foot drop in neuromuscular disease
20 - (FASEB, 21, 2007) Activation of PPAR stimulates utrophin A expression in skeletal muscle cells
A potential therapeutic strategy to treat Duchenne
Muscular Dystrophy (DMD) involves upregulation of utrophin in muscle
fibers of patients. Our recent studies demonstrated that utrophin
A is regulated by mechanisms that also promote expression of
the slow myofiber program (PNAS 100: 7791–6, 2003
20 - (FASEB, 21,
2007) The membrane sealant poloxamer
reduces membrane permeability in tibialis anterior muscles from dystrophic
Duchenne muscular dystrophy is characterised by the loss
of the membrane stabilizing protein dystrophin, resulting in an
increased susceptibility to sarcolemmal injury. The membrane
sealant Poloxamer 188 (P-188) prevents sarcolemmal tearing in cardiac
myocytes from dystrophic
20 - (FASEB, 21, 2007) IGF-I improves excitation-contraction coupling in skeletal muscle fibers of dystrophic mdx mice
The absence of dystrophin in Duchenne muscular dystrophy (DMD) causes progressive muscle wasting and possibly aberrant intracellular signaling including abnormal excitation-contraction coupling (ECC) in skeletal muscle. Insulin-like growth factor-I (IGF-I) is a potential therapy for DMD, and can alter ECC. We tested the hypothesis that muscle specific over-expression of IGF-I could ameliorate disruptions in ECC in muscle fibers from transgenic mdx (IGF-mdx) mice. Mechanically-skinned single fibers were prepared from the EDL muscles of mdx, IGF-mdx, and wild type mice. The number of depolarization-induced contractions (DICR) before a 50% reduction in amplitude was lower in mdx fibres (7 ± 1, n=15 fibers) than control (16 ± 2, n=12, P<0.05). There was no difference in SR Ca2+ properties. Rundown of DICR was improved significantly in IGF-mdx (14 ± 2, n=14) compared with mdx mice (7 ± 1, n=15, P<0.05). SR Ca2+ release was 300% higher in IGF-I-mdx mice, possibly due to a decrease in SR Ca2+ leak rate [mdx, 19.0 ± 3.5% of loaded SR (n=13) vs. IGF-mdx, 7.2 ± 3.0% (n=14)]. These results demonstrate that muscle specific over-expression of IGF-I in mdx mice can attenuate abnormal E-C coupling.
20 - (FASEB, 21, 2007) Roles of AML1 in Duchenne muscular dystrophy and in myogenesis
AML1 (acute myelogenous leukemia 1) was originally cloned
from the breakpoint of chromosome 21 in t(8;21)(q22;q22), and is
one of the most frequent targets of leukemia-associated gene
aberrations. AML1 is a transcription regulator of a number of
haematopoietic genes. The function of AML1 in skeletal muscle is not
clear. Based on expression profiling data of muscles of
non-symptomatic DMD patients (10m infants), symptomatic patients
(5–12yr), and
20 - (FASEB, 21, 2007) Disruption of IGF-I/IGFBP interactions decreases contraction-induced injury in dystrophic skeletal muscle
Duchenne muscular dystrophy (DMD) is caused by a lack of
dystrophin, a structural protein thought to confer stability to
skeletal muscle fibers. A lack of dystrophin results in an increased
susceptibility to contraction-induced (CI) injury. Insulin-like
growth factor-I (IGF-I) therapy has beneficial effects on muscle
function in
20 - (FASEB, 21, 2007) Local expression of IGF-1 accelerates muscle regeneration by rapidly modulating inflammatory cytokines and chemokines
Muscle regeneration following injury is characterized by myonecrosis accompanied by local inflammation, activation of satellite cells, and repair of injured fibers. The resolution of the inflammatory response is necessary to proceed toward muscle repair, since persistence of inflammation often renders the damaged muscle incapable of sustaining efficient muscle regeneration. Here, we show that local expression of a muscle-restricted insulin-like growth factor (IGF)-1 (mIGF-1) transgene accelerates the regenerative process of injured skeletal muscle, modulating the inflammatory response, and limiting fibrosis. At the molecular level, mIGF-1 expression significantly down-regulated proinflammatory cytokines, such as tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta, and modulated the expression of CC chemokines involved in the recruitment of monocytes/macrophages. Analysis of the underlying molecular mechanisms revealed that mIGF-1 expression modulated key players of inflammatory response, such as macrophage migration inhibitory factor (MIF), high mobility group protein-1 (HMGB1), and transcription NF-B. The rapid restoration of injured mIGF-1 transgenic muscle was also associated with connective tissue remodeling and a rapid recovery of functional properties. By modulating the inflammatory response and reducing fibrosis, supplemental mIGF-1 creates a qualitatively different environment for sustaining more efficient muscle regeneration and repair.
20 - (FASEB, 21, 2007) Physical activity and muscle adaptations in dystrophic mice treated with prednisone
Glucocorticoid administration is standard treatment for
individuals with Duchenne’s Muscular Dystrophy. However, the drug
effects on physical activity and skeletal muscle are not clearly
established. We tested the hypothesis that
20 - (FASEB, 21, 2007) A calpain inhibitor fails to rescue dystrophic skeletal muscle
Duchenne muscular dystrophy (DMD) is a fatal muscle
disease caused by the absence of a functional dystrophin protein and
is modeled by the
20 - (FASEB, 21,
2007)
Chemical sympathectomy ameliorates the dystrophic
pathology of limb and diaphragm muscles in
Mutations of the dystrophin gene cause Duchenne muscular
dystrophy, but the pathogenetic mechanisms are poorly understood. We
previously showed that vasoconstriction evoked by activation of
sympathetic nerves or vascular
-adrenoceptors
is enhanced in dystrophin-deficient muscles during exercise. We
therefore hypothesized that sympathetically-mediated muscle ischemia
might contribute to the dystrophic phenotype. To test this, we
treated C57BL10 and
Genome-wide gene expression profiling of skeletal muscle from Duchenne muscular dystrophy (DMD) patients has been used to describe muscle tissue alterations in DMD children older than 5 years. By studying the expression profile of 19 patients younger than 2 years, we describe with high resolution the gene expression signature that characterizes DMD muscle during the initial or "presymptomatic" phase of the disease. We show that in the first 2 years of the disease, DMD muscle is already set to express a distinctive gene expression pattern considerably different from the one expressed by normal, age-matched muscle. This "dystrophic" molecular signature is characterized by a coordinate induction of genes involved in the inflammatory response, extracellular matrix (ECM) remodeling and muscle regeneration, and the reduced transcription of those involved in energy metabolism. Despite the lower degree of muscle dysfunction experienced, our younger patients showed abnormal expression of most of the genes reported as differentially expressed in more advanced stages of the disease. By analyzing our patients as a time series, we provide evidence that some genes, including members of three pathways involved in morphogenetic signaling—Wnt, Notch, and BMP—are progressively induced or repressed in the natural history of DMD.
20 - (FASEB, 21, 2007) The brain-derived neurotrophic factor (BDNF) regulates skeletal muscle regeneration and is mis-regulated in dystrophic muscle
Our recent studies have shown that BDNF is highly
expressed in skeletal muscle satellite cells (J.
Neurosci., 26: 5739, 2006
20 - Myocardial Delayed Enhancement by Magnetic Resonance Imaging in Patients With Muscular Dystrophy
Objectives: This study sought to analyze whether cardiovascular magnetic resonance (CMR) can detect and quantify myocardial damage in the early stages of cardiomyopathy in muscular dystrophies (MD).
Background: Muscular dystrophy is a genetic disease that involves skeletal and cardiac tissues of humans. Cardiomyopathy is common, and death secondary to cardiac or respiratory diseases occurs early in life. Cardiovascular magnetic resonance is a reliable method for assessing global and regional cardiac function, allowing also for the detection of myocardial fibrosis (MF).
Methods: Ten patients with Duchenne or Becker dystrophies were studied by CMR. Physical examination, Chagas disease serological tests, electrocardiogram, chest radiograph, total creatine kinase, and Doppler echocardiogram were also obtained in all patients.
Results: Patients with MF had a lower ejection fraction than those without. Myocardial fibrosis (midwall and/or subepicardial) was observed in 7 of the 10 patients, and the lateral wall was the most commonly involved segment. There was moderate correlation between segmental MF and dysfunction.
Conclusions: Cardiovascular magnetic resonance can identify MF and may be useful for detecting the early stages of cardiomyopathy in MD. Future work will be needed to evaluate whether CMR can influence cardiomyopathy and outcomes.
19 - (FASEB, 21, 2007) Reactive oxygen species promote sarcolemmal damage following ischemia and reperfusion of dystrophin-deficient muscles
Mutations of the dystrophin gene cause Duchenne muscular dystrophy, but the pathogenetic mechanisms are poorly understood. We previously reported that dystrophin-deficient muscles are susceptible to ischemia during exercise. Reactive oxygen species (ROS) produced during reperfusion of ischemic muscle can cause tissue injury. Because dystrophin-deficient muscle is abnormally sensitive to ROS, we hypothesized that ischemia-reperfusion (I/R) injury would be enhanced in dystrophin-deficient muscles of mdx mice, and that this damage would be ameliorated by antioxidant treatment. Anesthetized mdx and C57BL10 mice were subjected to 1 h of unilateral hindlimb ischemia followed by 2 h of reperfusion. Sarcolemmal damage following I/R was enhanced in mdx vs C57BL10 mice as shown by greater increases in serum creatine kinase (+2775±260 vs +1136±211 U/L; P<0.05) and in Evans blue dye (EBD) uptake by muscle (EBD+ area: 31±6 vs 8±3% of plantaris; 27±5 vs 3±1% of soleus; P<0.05). Infusion of the superoxide dismutase mimetic tempol (172 mg/kg, iv) attenuated EBD uptake by 63% in plantaris and by 31% in soleus muscles (P<0.05 vs I/R alone) of mdx mice, but had no effect in C57BL10 mice. These data demonstrate that the hindlimb muscles of mdx mice are abnormally susceptible to I/R-induced injury, and suggest that ROS production is a key event mediating sarcolemmal damage in dystrophin-deficient muscle.
18 - (Biochemical and Biophysical Research Communications, 2007) Efficient conversion of ES cells into myogenic lineage using the gene-inducible system
Shiro Ozasa, Shigemi Kimura, Kaori Ito, Hiroe Ueno, Makoto Ikezawa, Makoto Matsukura, Kowashi Yoshioka, Kimi Araki, Ken-ich Yamamura, Kuniya Abe, Hitoshi Niwa and Teruhisa Miike - Japan
We established genetically engineered ES (ZHTc6-MyoD) cells that harbor a tetracycline-regulated expression vector encoding myogenic transcriptional factor MyoD, for the therapy of muscle diseases, especially Duchenne muscular dystrophy (DMD). Almost all the ZHTc6-MyoD cells were induced into muscle lineage after removal of tetracycline. The undifferentiated ZHTc6-MyoD cells are Sca-1+ and c-kit+, but CD34−, all well-known markers for mouse hematopoietic stem cells. In addition, they are able to maintain themselves in the undifferentiated state, even after one month of culture. Therefore, it is possible to obtain a large quantity of ZHTc6-MyoD cells in the undifferentiated state that maintain the potential to differentiate only into muscle lineage. Additionally, at two weeks post-injection of these cells into muscle of mdx, a model mouse of DMD, clusters of dystrophin-positive myofibers were observed at the injection site. Therefore, ES cells have considerable therapeutic potential for treating muscle diseases.
13 - Evaluation of narrative abilities in patients suffering from Duchenne Muscular Dystrophy
4 - (J Appl Physiol 102: 1677–1686, 2007) The role of free radicals in the pathophysiology of muscular dystrophy
James G. Tidball and Michelle Wehling-Henricks - USA
Null mutation of any one of several members of the dystrophin protein complex can cause progressive, and possibly fatal, muscle wasting. Although these muscular dystrophies arise from mutation of a single gene that is expressed primarily in muscle, the resulting pathology is complex and multisystemic, which shows a broader disruption of homeostasis than would be predicted by deletion of a single-gene product. Before the identification of the deficient proteins that underlie muscular dystrophies, such as Duchenne muscular dystrophy (DMD), oxidative stress was proposed as a major cause of the disease. Now, current knowledge supports the likelihood that interactions between the primary genetic defect and disruptions in the normal production of free radicals contribute to the pathophysiology of muscular dystrophies. In this review, we focus on the pathophysiology that results from dystrophin deficiency in humans with DMD and the mdx mouse model of DMD. Current evidence indicates three general routes through which free radical production can be disrupted in dystrophin deficiency to contribute to the ensuing pathology. First, constitutive differences in free radical production can disrupt signaling processes in muscle and other tissues and thereby exacerbate pathology. Second, tissue responses to the presence of pathology can cause a shift in free radical production that can promote cellular injury and dysfunction. Finally, behavioral differences in the affected individual can cause further changes in the production and stoichiometry of free radicals and thereby contribute to disease. Unfortunately, the complexity of the free radical mediated processes that are perturbed in complex pathologies such as DMD will make it difficult to develop therapeutic approaches founded on systemic administration of antioxidants. More mechanistic knowledge of the specific disruptions of free radicals that underlie major features of muscular dystrophy is needed to develop more targeted and successful therapeutic approaches.
4 - Osteoporosis in children with neuromuscular diseases and inborn errors of metabolism
MARCH
31 - Switching genes to overdrive improves muscular dystrophy symptoms in mice
27 - (American College of Cardiology Meeting 2007) Cardiac Myolysis And LV Dysfunction In Patients With Duchenne Muscular Dystrophy -Inhibitory Effect Of Carvedilol On Cardiac Myolysis
Takao Nishizawa, Fumihiko Yasuma, Akiko Noda, Hideo Izawa, Kohzo Nagata,Mitsuhiro Yokota, Toyoaki Murohara - Japan
Background: Cardiac myolysis is observed in idiopathic dilated cardiomyopathy (DCM) and elevation of a biochemical marker leads to poor prognosis. Patients with Duchenne muscular dystrophy (DMD) often have cardiomyopathy. However it is unclear that how cardiac myolysis occurs in patients with DMD and how it relates to cardiac function.
Methods: Fifty-eight Duchenne muscular dystrophy patients were subjected of this study. Serum CPK and plasma cardiac troponin I (cTnI) were repeatedly (every 3 months) measured for 2 years and LV function was assessed by echocardiography.
Results: Patients with positive cTnI (Cut off 0.06ng/mL) showed lower LVEF(42± 2%) than those of negative cTnI (52± 2%). However, 28% of the impaired LV function (LVEF<50%) never showed positive cTnI. The frequency of detecting positive cTnI was decreased according to age and decreased serum CPK level. Some patients in less than 20 years old showed positive cTnI with preserved LV function. On the other hand, patients with preserved LV function in more than 20 years old never showed positive cTnI. Administration in carvedilol (2.5-5 mg/day) in 12 patients with positive cTnI for 6 months decreased the frequency of positive cTnI (From 100% to 25%).
Conclusions: Plasma cTnI was associated with impaired LV function. Positive plasma cTnI precedes the impaired cardiac function and the frequency of detecting plasma cTnI decreased with decreased LVEF suggesting that persistent myolysis become intermittent as progression of LV dysfunction. Repeated measurements of plasma cTnI may be useful to predict development of LV dysfunction. Carvedilol administration is one of the effective therapies to prevent cardiac myolysis.
26 - (J. Clin. Pharmacol,2007; 47:430-444) Safety, Tolerability, and Pharmacokinetics of PTC124, a Nonaminoglycoside Nonsense Mutation Suppressor, Following Single- and Multiple-Dose Administration to Healthy Male and Female Adult Volunteers
Nonsense (premature stop codon) mutations are causative in 5% to 15% of patients with monogenetic inherited disorders. PTC124, a 284-Dalton 1,2,4-oxadiazole, promotes ribosomal readthrough of premature stop codons in mRNA and offers therapeutic potential for multiple genetic diseases. The authors conducted 2 phase I studies of PTC124 in 62 healthy adult volunteers. The initial, single-dose study evaluated doses of 3 to 200 mg/kg and assessed fed-fasting status on pharmacokinetics following a dose of 50 mg/kg. The subsequent multiple-dose study evaluated doses from 10 to 50 mg/kg/dose twice per day (bid) for up to 14 days. PTC124 administered orally as a liquid suspension was palatable and well tolerated through single doses of 100 mg/kg. At 150 and 200 mg/kg, PTC124 induced mild headache, dizziness, and gastrointestinal events. With repeated doses through 50 mg/kg/dose bid, reversible transaminase elevations <2 times the upper limit of normal were sometimes observed. Immunoblot analyses of peripheral blood mononuclear cell extracts revealed no protein elongation due to nonspecific ribosomal readthrough of normal stop codons. PTC124 plasma concentrations exceeding the 2- to 10-µg/mL values associated with activity in preclinical genetic disease models were safely achieved. No sex-related differences in pharmacokinetics were seen. No drug accumulation with repeated dosing was apparent. Diurnal variation was observed, with greater PTC124 exposures after evening doses. PTC124 excretion in the urine was <2%. PTC124 pharmacokinetics were described by a 1-compartment model. Collectively, the data support initiation of phase II studies of PTC124 in patients with nonsense mutation–mediated cystic fibrosis and Duchenne muscular dystrophy.
23 - (J. Clin. Invest,2007) Interplay of IKK/NF-B signaling in macrophages and myofibers promotes muscle degeneration in Duchenne muscular dystrophy
Duchenne muscular dystrophy (DMD) is a lethal X-linked disorder associated with dystrophin deficiency that results in chronic inflammation and severe skeletal muscle degeneration. In DMD mouse models and patients, we find that IB kinase/NF-B (IKK/NF-B) signaling is persistently elevated in immune cells and regenerative muscle fibers. Ablation of 1 allele of the p65 subunit of NF-B was sufficient to improve pathology in mdx mice, a model of DMD. In addition, conditional deletion of IKKß in mdx mice elucidated that NF-B functions in activated macrophages to promote inflammation and muscle necrosis and in skeletal muscle fibers to limit regeneration through the inhibition of muscle progenitor cells. Furthermore, specific pharmacological inhibition of IKK resulted in improved pathology and muscle function in mdx mice. Collectively, these results underscore the critical role of NF-B in the progression of muscular dystrophy and suggest the IKK/NF-B signaling pathway as a potential therapeutic target for DMD.
More informations about IKK/NF-kB:
"Since NF-κB regulation maintains a delicate balance
between cell survival and apoptosis and is very important for host defense
mechanisms, the specificity and duration of the NF-κB–inhibiting agent becomes
highly relevant in designing effective pharmacological therapies against chronic
diseases. Whereas IKKβ inhibition in macrophages is beneficial in reducing
inflammation in cancer, IKKα loss in macrophages can actually elevate the immune
response by increasing the production of proinflammatory cytokines and
chemokines, thus underscoring the importance of specificity for IKK inhibitors.
Similarly, though administration of nonspecific agents such as MG-132 and,
more recently, a synthetic antioxidant ,has been shown to ameliorate the
dystrophic phenotype, risks of inhibiting unwanted cellular targets can also
sometimes outweigh the benefits. In addition, our inability to detect clear
evidence of IκBα degradation in dystrophic muscles may argue against the use of
general proteasome inhibitors as a therapeutic strategy for blocking classical
NF-κB signaling in DMD. Hence, the implementation of a specific NF-κB
inhibitor such as the NBD peptide might provide maximal therapeutic benefit with
minimal side effects (. Although long-term inhibition of NF-κB is
associated with a vulnerability to bacterial infections and mounting immune
responses, it is noteworthy that the NBD peptide has not been found to block
basal NF-κB activity and, as seen in this and
previously described studies, does not exhibit overt signs of toxicity.
Together, the results identify the IKK/NF-κB signaling
pathway as a viable target for anti-DMD therapy."
23 - (Am. J. Respir. Crit. Care Med. 2007) Leupeptin Inhibits Ventilator-Induced Diaphragm Dysfunction in Rats
Karen Maes,
Rationale: Controlled mechanical ventilation has been shown to result in elevated diaphragmatic proteolysis and atrophy along with diaphragmatic contractile dysfunction. Objectives: To test whether administration of leupeptin, an inhibitor of lysosomal proteases and calpain, concomitantly with 24 hours of controlled mechanical ventilation would protect the diaphragm from the deleterious effects of mechanical ventilation. Methods: Rats were assigned to either a control group or 24 hours of controlled mechanical ventilation; animals in the ventilation group received either a single intramuscular injection of saline or 15 mg/kg of the protease inhibitor leupeptin. Measurements and main results: Compared with controls, mechanical ventilation resulted in a significant reduction of the in vitro diaphragm specific force production at all stimulation frequencies. Leupeptin completely prevented this reduction in force generation. Atrophy of type IIx/b fibers was present following controlled mechanical ventilation but not after treatment with leupeptin. Cathepsin B and calpain activities were significantly higher following controlled mechanical ventilation compared to the other groups; this was abolished by treatment with leupeptin. Significant inverse correlations were found between diaphragmatic force generation and cathepsin B and calpain activity and illustrate the deleterious role of proteolysis in diminishing diaphragmatic force production following prolonged controlled mechanical ventilation. Conclusion: Administration of the protease inhibitor leupeptin concomitantly with mechanical ventilation completely prevented ventilation-induced diaphragmatic contractile dysfunction and atrophy.
20 - (Am
J Phys Med Rehabil 2007;86:295-300)
Lung Inflation by Glossopharyngeal Breathing and "Air Stacking" in Duchenne
Muscular
Dystrophy.
Bach JR, Bianchi C, Vidigal-Lopes M, Turi S, Felisari G - USA
Objective: To compare the use of glossopharyngeal breathing (GPB) and air
stacking to increase lung volumes and cough peak flows (CPF), and GPB to
increase ventilator-free breathing ability (VFBA), for patients with Duchenne
muscular
dystrophy.
Design: A case series of all referred patients with declining vital capacity (VC).
Seventy-eight patients underwent training in and monitoring of the efficacy of
air stacking (retaining consecutively delivered volumes of air delivered via
manual resuscitator and held by glottic closure) to maximum insufflation
capacity (MIC). GPB also was demonstrated to all 78 patients, and 32 were
formally trained and prescribed GPB as their VCs decreased below 400 ml. To be
successful, the MIC or GPB maximum single-breath capacity (GPmaxSBC) had to
exceed VC. Improvements in VFBA were determined by requiring fewer
ventilator-assisted breaths per minute. CPFs were measured by peak flow meter.
Results: Seventy-four (94.9%) of the patients could air stack (MIC > VC), and,
thus far, 21 (27%) are able to GPB. Fifteen could GPB sufficiently to delay
onset of daytime ventilator use and, later, to require 1.9 fewer ventilator
assisted breaths per minute. For the 47 patients with multiple data points, as
VC deteriorated from 1080 +/- 870 to 1001 +/- 785 ml, MIC increased from 1592
+/- 887 to 1838 +/- 774 ml. For 21 patients, GPmaxSBC significantly exceeded VC
(824 +/- 584 vs. 244 +/- 151 ml, respectively, P < 0.001). The ability to
increase lung volume by air stacking (MIC) was better retained than was the
ability to increase lung volume by GPB (GPmaxSBC). Air stacking also permitted
assisted CPF to exceed unassisted CPF: 289 +/- 91 and 164 +/- 76 liters/m,
respectively (P < 0.001).
Conclusions: GPB and air stacking can increase lung volumes and, thereby, cough
flows. GPB also can be used in many cases to delay and decrease daytime
ventilator use.
14 - BM stem cell transplantation rescues pathophysiologic features of aged dystrophic mdx muscle
03 - (Neuromuscular Disorders, 2007) Myostatin does not regulate cardiac hypertrophy or fibrosis
Ronald D. CohN, Hsin-Yueh Liang, Reena Shetty, Theodore Abraham and Kathryn R. WagneR - USA
Myostatin is a negative regulator of muscle growth. Loss of myostatin has been shown to cause increase in skeletal muscle size and improve skeletal muscle function and fibrosis in the dystrophin-deficient mdx muscular dystrophy mouse model. We evaluated whether lack of myostatin has an impact on cardiac muscle growth and fibrosis in vivo. Using genetically modified mice we assessed whether myostatin absence induces similar beneficial effects on cardiac function and fibrosis. Cardiac mass and ejection fraction were measured in wild-type, myostatin-null, mdx and double mutant mdx/myostatin-null mice by high resolution echocardiography. Heart mass, myocyte area and extent of cardiac fibrosis were determined post mortem. Myostatin-null mice do not demonstrate ventricular hypertrophy when compared to wild-type mice as shown by echocardiography (ventricular mass 0.69 ± 0.01 vs. 0.69 ± 0.018 g) and morphometric analyses including heart/body weight ratio (5.39 ± 0.45 vs. 5.62 ± 0.58 mg/g) and cardiomyocyte area 113.67 ± 1.5, 116.85 ± 1.9 μm2). Moreover, absence of myostatin does not attenuate cardiac fibrosis in the dystrophin-deficient mdx mouse (12.2% vs. 12%). The physiological role of myostatin in cardiac muscle appears significantly different than that in skeletal muscle as it does not induce cardiac hypertrophy and does not modulate cardiac fibrosis in mdx mice.
03 -
Multinational Trial to Evaluate Steroids in DMD
FEBRUARY
24 - Poloxamer 188 failed to prevent exercise-induced membrane breakdown in mdx skeletal muscle fibers
24 - Streptomycin reduces stretch-induced membrane permeability in muscles from mdx mice.
22 - (Molecular Biology Cell, 2007) Menstrual Blood-derived Cells Confer Human Dystrophin Expression in the Murine Model of Duchenne Muscular Dystrophy via Cell Fusion and Myogenic Transdifferentiation
ChangHao Cui,
Duchenne muscular dystrophy, the most common lethal genetic disorder in children, is an X-linked recessive muscle disease characterized by the absence of dystrophin at the sarcolemma of muscle fibers. We examined a putative endometrial progenitor obtained from endometrial tissue samples to determine whether these cells repair muscular degeneration in a murine mdx model of duchenne muscular dystrophy. Implanted cells conferred human dystrophin in degenerated muscle of immunodeficient mdx mice. We then examined menstrual blood-derived cells to determine whether primarily cultured nontransformed cells also repair dystrophied muscle. In vivo transfer of menstrual blood-derived cells into dystrophic muscles of immunodeficient mdx mice restored sarcolemmal expression of dystrophin. Labeling of implanted cells with EGFP and differential staining of human and murine nuclei suggest that human dystrophin expression is due to cell fusion between host myocytes and implanted cells. In vitro analysis revealed that endometrial progenitor cells and menstrual blood-derived cells can efficiently transdifferentiate into myoblasts/myocytes, fuse to C2C12 murine myoblasts by in vitro coculturing, and start to express dystrophin after fusion. These results demonstrate that the endometrial progenitor cells and menstrual blood-derived cells can transfer dystrophin into dystrophied myocytes at a high frequency through cell fusion and transdifferentiation in vitro and in vivo.
17 - Intracellular calcium handling in ventricular myocytes from mdx mice
15 - (Neuromuscular Disorders, 2007) The allure of stem cell therapy for muscular dystrophy
Miranda D. Grounds , Kay E. Davies - Australia and UK
Duchenne muscular dystrophy (DMD) is a lethal muscle disease for which an effective treatment is urgently needed. The use of stem cells to produce normal muscle cells to replace the missing dystrophin protein has attracted much attention. Claims of success using stem cell treatment in animal models of human muscle diseases require careful evaluation and are not necessarily easily extrapolated to the clinical situation. Recent studies in the dystrophic dog model have been claimed to show that injected mesangioblasts, stem cells derived from blood vessels, reduce the severity of the disease. However, the authors’ interpretation of the results did not consider that benefits might arise from the concomitant use of immunosuppressive drugs alone.
15 - (Neuromuscular Disorders, 2007) Serial casting of the ankles in Duchenne muscular dystrophy: can it be an alternative to surgery?
Marion Main, Eugenio Mercuri, Goknur Haliloglu, Ros Bake, Maria Kinali, Francesco Muntoni - UK
The aim of this non-randomized pilot study was to assess whether serial casting can be used in Duchenne muscular dystrophy (DMD) patients to reduce tendo-achilles (TA) contractures and to allow the fitting of the knee–ankle–foot orthoses (KAFOs) at the end of the functional walking. TA range and age at loss of standing with KAFOs were prospectively assessed in nine DMD patients who underwent serial casting prior to rehabilitation into KAFOs and compared to a group of 20 DMD patients who had TA (group II) or TA and iliotibial band (ITB) surgical resection (group III). TA range after casting was reduced at least to plantargrade/neutral in 8 of the 9 patients studied although patients who had surgery had better correction of TA angles. The mean interval between rehabilitation and loss of standing ability with KAFOs was only minimally different among the three groups 1.8 years (9 months–2.8 years) in group I, 2 years (1 month–4.3 years) in group II, 1.9 years (6 months–4.7 years) in group III. The results of this pilot study suggest that serial casting of the TAs can be considered in DMD patients with moderate TA contractures and in whom there are no significant iliotibial band (ITB) tightness, although this intervention is probably not as effective as surgery in maintaining long-term TA range. This procedure may therefore be considered both in patients with medical contraindication to surgery, and also in patients or families who are not prepared to cope with the stress and discomfort of TA surgery.
15 - (SPINE Volume 32, Number 4, pp 459–465) Posterior Spinal Fusion for Scoliosis in Duchenne Muscular Dystrophy Diminishes the Rate of Respiratory Decline
Maria V. Velasco, Andrew A. Colin,, David Zurakowski, Basil T. Darras, and Frederic Shapiro - USA
Study Design. To assess the rate of decline in pulmonary function in Duchenne muscular dystrophy (DMD) before and after posterior spinal fusion for scoliosis.
Objective. To compare the rate of respiratory decline using percent normal forced vital capacity (%FVC) measurements before and after posterior spinal fusion.
Summary of Background Data. Posterior spinal fusion for scoliosis is used widely in DMD, although the long term pulmonary effects have not been well established.
Methods. Fifty-six patients were assessed. Percent forced vital capacity was the outcome parameter with data analysis using a mixed-model repeated-measures ANOVA and paired t tests. Group 1: Inclusion criteria were a diagnosis of DMD, 2 or more pulmonary function tests presurgery, and 2 or more postsurgery. Group 2: The rates of respiratory decline before and after spinal fusion for the whole study population were determined by withinsubjects mixed-model regression analysis to account for the varying number of FVC studies between patients and unequal spacing between tests.
Results. Group 1: 20 patients. Mean length of time of respiratory value determination was 2.5 + 1.0 years presurgery and 5.6 + 2.8 years postsurgery. Mean rate of decline presurgery was 8.0% + 4.1% per year, which decreased to 3.9% + 1.9% per year postsurgery (paired t test 4.58, P < 0.0001). Group 2: 56 patients. The respiratory value determinations ranged from 4 years presurgery to 8 years postsurgery. The rates of respiratory decline based on the whole study population were 4% per year presurgery, which decreased to 1.75% per year postsurgery (F-test comparison of slopes 19.71, P < 0.0001).
Conclusions. Posterior spinal fusion for scoliosis in DMD is associated with a significant decrease in the rate of respiratory decline postsurgery compared with presurgery rates.
12 - (Nature Cell Biology , 2007) Pericytes of human skeletal muscle are myogenic precursors distinct from satellite cells
Arianna Dellavalle, Maurilio Sampaolesi
, Rossana Tonlorenzi, Enrico Tagliafico, Benedetto Sacchetti, Laura Perani, Anna Innocenzi, Beatriz G. Galvez, Graziella Messina, Roberta Morosetti, Sheng Li, Marzia Belicchi, Giuseppe Peretti, Jeffrey S. Chamberlain, Woodring E. Wright, Yvan Torrente, Stefano Ferrari, Paolo Bianco and Giulio Cossu - ItalyCells derived from blood vessels of human skeletal muscle can regenerate skeletal muscle, similarly to embryonic mesoangioblasts. However, adult cells do not express endothelial markers, but instead express markers of pericytes, such as NG2 proteoglycan and alkaline phosphatase (ALP), and can be prospectively isolated from freshly dissociated ALP+ cells. Unlike canonical myogenic precursors (satellite cells), pericytederived cells express myogenic markers only in differentiated myotubes, which they form spontaneously with high efficiency. When transplanted into severe combined immune deficient–Xlinked, mouse muscular dystrophy (scid–mdx) mice, pericytederived cells colonize host muscle and generate numerous fibres expressing human dystrophin. Similar cells isolated from Duchenne patients, and engineered to express human minidystrophin, also give rise to many dystrophin-positive fibres
in vivo. These data show that myogenic precursors, distinct from satellite cells, are associated with microvascular walls in the human skeletal muscle, may represent a correlate of embryonic ‘mesoangioblasts’ present after birth and may be a promising candidate for future cell-therapy protocols in patients.10 - 20 Years after finding the Duchenne Gene: A Terrible Disease is being Conquered
10 - (Chinese Journal of Biotechnology, 2007; 23(1):27-32) Construction of Recombinant Adenovirus Including Microdystrophin and Expression in the Mesenchymal Cells of mdx Mice
Fu XIONG, Cheng ZHANG, Shao-Bo XIAO, Mei-Shan LI, Shu-Hui WANG, Mei-Juan YU and Yan-Chang SHANG
Construction of recombinant adenovirus, which contain human microdystrophin, and then transfection into mesenchymal cells (MSCs) of mdx mice were done, and genetically-corrected isogenic MSCs were acquired; the MSCs transplantation into the mdx mice was then done to treat the Duchenne muscular dystrophy (DMD). Microdystrophin cDNA was obtained from recombinant plasmid pBSK-MICRO digested with restrictive endonuclease Not I; the production was inserted directionally into pShuttle-CMV. The plasmid of pShuttle-CMV-MICRO was digested by Pme I, and the fragment containing microdystrophin was reclaimed and transfected into E. coli BJ5183 with plasmid pAdeasy-1. After screening by selected media, the extracted plasmid of positive bacteria was transfected into HEK293 cells with liposome and was identified by observing the CPE of cells and by the PCR method. Finally, MSCs of mdx mice were infected with the culture media containing recombinant adenovirus, and the expression of microdystrophin was detected by RT-PCR and immunocytochemistry. Recombinant adenovirus including microdystrophin was constructed successfully and the titer of recombinant adenovirus was about 5.58 × 1012 vp/mL. The recombinant adenovirus could infect MSC of mdx mice and microdystrophin could be expressed in the MSC of mdx mice. Recombinant adenovirus including microdystrophin was constructed successfully, and the microdystrophin was expressed in the MSC of mdx mice. This lays the foundation for the further study of microdystrophin as a target gene to correct the dystrophin-defected MSC for stem cell transplantation to cure DMD.
10 -
Antisense Oligonucleotide-induced Exon Skipping Across the Human Dystrophin Gene
Transcript
7 - (In Press:
Chest, 2007; 131,368-75)
Lung Function Accurately Predicts Hypercapnia
in Patients With Duchenne Muscular Dystrophy
Michel Toussaint, Marc Steens, and Philippe Soudon - Belgium
Background:
In patients with Duchenne muscular dystrophy (DMD), implementation of mechanical ventilation depends on sleep investigation and measurement of CO2 tension. The objective of this cross-sectional study was to determine which noninvasive lung function parameter best predicts nocturnal hypercapnia and diurnal hypercapnia in these patients.Methods: According to transcutaneous CO2 (TcCO2) measurement, 114 DMD patients were classified into three groups: nocturnal hypercapnia (n = 38) [group N], diurnal hypercapnia (n = 39), despite nocturnal ventilation (group D), and 24-h normocapnia and spontaneous breathing (n = 37) [group S] as control. TcCO2 tension and lung function variables included vital capacity (VC) and maximal inspiratory pressure (MIP), and breathing pattern variables included tidal volume (VT) and respiratory rate (RR), measured at the time of group inclusion. The rapid and shallow breathing index (RSBI [RR/VT]) and VT/VC ratio were calculated. Areas under the curve from the receiver operating characteristic (ROC) were calculated for those parameters.
Results: Compared to group S, lung function was significantly worse in group N and group D. VC, RR, and RSBI distinguished group S from group N by ROC comparison. Cut-off values of VC < 680 mL (ROC, 0.968), MIP < 22 cm H2O (ROC, 0.928), and VT/VC > 0.33 (ROC, 0.923) accurately discriminated group D from group N, but RSBI, RR, and VT did not.
Conclusions: Lung function is useful to predict nocturnal hypercapnia in patients with DMD. Moreover, VC < 680 mL is very sensitive to predict daytime hypercapnia.
3 - (In Press: Neuropathology and Applied Neurobiology, 2007) Role of tumour necrosis factor alpha, but not of cyclo-oxygenase-2-derived eicosanoids, on functional and morphological indices of dystrophic progression in mdx mice: a pharmacological approach
S. Pierno, B. Nico, R. Burdi, A. Liantonio, M. P. Didonna, V. Cippone, B. Fraysse, J.-F. Rolland, D. Mangieri, Andreetta, P. Ferro, C. Camerino, A. Zallone, P. Confalonieri and A. De Luca
The role of tumour necrosis factor (TNF)-alpha or cyclooxygenase- 2 (COX-2) eicosanoids in dystrophinopathies has been evaluated by chronically treating (4–8 weeks) adult dystrophic mdx mice with the anti-TNF-alpha etanercept (0.5 mg/kg) or the COX-2 inhibitor meloxicam (0.2 mg/kg). Throughout the treatment period the mdx mice underwent a protocol of exercise on treadmill in order to worsen the pathology progression; gastrocnemious muscles from exercised mdx mice showed an intense staining for TNF-alpha by immunohistochemistry. In vivo, etanercept, but not meloxicam, contrasted the exercise induced forelimb force drop. Electrophysiological recordings ex vivo, showed that etanercept counteracted the decrease in chloride channel function (gCl), a functional index of myofibre damage, in both diaphragm and extensor digitorum longus (EDL) muscle, meloxicam being effective only in EDL muscle. None of the drugs ameliorated calcium homeostasis detected by electrophysiology and/or spectrofluorimetry. Etanercept, more meloxicam, effectively reduced plasma creatine (CK). Etanercept-treated muscles showed a reduction connective tissue area and of pro-fibrotic cytokine vs. untreated ones; however, the histological profile weakly ameliorated. In order to better evaluate the of etanercept treatment on histology, a 4-week treatment was performed on 2-week-old mdx mice, so to match first spontaneous degeneration cycle. The histology of gastrocnemious was significantly improved reduction of degenerating area; however, CK levels only slightly lower. The present results support a key of TNF-alpha, but not of COX-2 products, in different phases dystrophic progression. Anti-TNF-alpha drugs may be in combined therapies for Duchenne patients.
1 - Efficient Whole-body Transduction with Trans-splicing Adeno-associated Viral Vectors
JANUARY
31 - Lack of myostatin results in excessive muscle growth but impaired force generation
27 - Verbal and memory skills in males with Duchenne muscular dystrophy
27 - Surgery for scoliosis in Duchenne muscular dystrophy
27 - Creatine for treating muscle disorders
21 - (In Press: Nature Medicine, 2007) Angiotensin II type 1 receptor blockade attenuates TGF-beta–induced failure of muscle regeneration in multiple myopathic states
Ronald D Cohn, Christel van Erp, Jennifer P Habashi, Arshia A Soleimani, Erin C Klein, Matthew T Lisi, Matthew Gamradt, Colette M ap Rhys, Tammy M Holm, Bart L Loeys, Francesco Ramirez, Daniel P Judge, Christopher W Ward & Harry C Dietz - USA
Skeletal muscle has the ability to achieve rapid repair in response to injury or disease. Many individuals with Marfan syndrome (MFS), caused by a deficiency of extracellular fibrillin-1, exhibit myopathy and often are unable to increase muscle mass despite physical exercise. Evidence suggests that selected manifestations of MFS reflect excessive signaling by transforming growth factor (TGF)-beta. TGF-beta is a known inhibitor of terminal differentiation of cultured myoblasts; however, the functional contribution of TGF-beta signaling to disease pathogenesis in various inherited myopathic states in vivo remains unknown. Here we show that increased TGF-beta activity leads to failed muscle regeneration in fibrillin-1– deficient mice. Systemic antagonism of TGF-beta through administration of TGF-beta–neutralizing antibody or the angiotensin II type 1 receptor blocker losartan normalizes muscle architecture, repair and function in vivo. Moreover, we show TGF-beta–induced failure of muscle regeneration and a similar therapeutic response in a dystrophin-deficient mouse model of Duchenne muscular dystrophy.
(It's the same research published in October/2006 in this website)
21 - (In Press: Clinical Biochemistry, 2007) Decreased calcineurin activity in circulation of Duchenne muscular dystrophy
J. Shanmuga Sundarama, V. Mohana Raoa, A.K. Meenab, and M.P.J.S.Anandaraja - India
Objectives: Analysis of enzymes in easily accessible tissues like blood cells, serum can provide valuable information and simple tool for disease and carrier detection. In the study presented we have analyzed calcineurin activity in Duchenne muscular dystrophy (DMD) and carrier sera and lymphocytes for its diagnostic value and its status in DMD pathology. Design and methods: We have monitored calcineurin activity in sera and lymphocytes of DMD, carriers and in controls using colorimetric method by following the p-nitrophenol released in the presence and absence of Trifluoperazine (TFP), an inhibitor of calcineurin. Results: Results showed a significant decrease in serum and lymphocyte calcineurin activity in DMD (p<0.001) without alteration in carriers compared to normal. Conclusion: Further studies are required to understand possible alterations mediated by calcineurin with reference to DMD lymphocytes as any alteration in phosphorylation / dephosphorylation pathway can disturb the normal functioning of these cells. The decreased calcineurin activity observed in DMD serum compared with controls could be further examined for its diagnostic utility.
16 - New Approaches in the Therapy of Cardiomyopathy in Muscular Dystrophy
13 - Do all patients with Duchenne muscular dystrophy require surgery for the correction of scoliosis?
BACKGROUND: Most patients with
Duchenne muscular
dystrophy (DMD) develop scoliosis,
which can compromise sitting posture and respiratory function. There is
controversy as to whether and when scoliosis should be corrected surgically.
OBJECTIVE: To evaluate the efficacy of an individualized approach to
managing scoliosis in DMD.
DESIGN AND INTERVENTION: In this retrospective study, the investigators
reviewed case notes on 123 boys with DMD who were at least 17 years old at the
time of the study and who had been evaluated at a British hospital for at least
10 years (1992-2002). At the loss of ambulation, patients were remobilized in
lightweight knee-ankle-foot orthoses. In the 90% of the study population who
developed scoliosis, sitting anteroposterior spinal radiographs were taken every
6 months. The Cobb method was used to determine the degree of the spinal
curvature. Patients with a Cobb angle above 20[degrees] were offered
thoracolumbar spinal orthoses, and patients with rapidly progressing scoliosis (progress
of at least 10[degrees] per month) or with severe scoliosis (Cobb angle >50[degrees])
were considered for surgery. Respiratory function was assessed every 6 months by
recording forced vital capacity (FVC). The investigators compared the outcome at
17 years of age between surgical candidates (n = 70) and conservatively managed
patients (n = 53). The surgical-candidate group comprised 43 patients who
underwent surgery, 11 patients who refused surgery and 16 patients with severe
scoliosis who were unfit for surgery because of poor cardiopulmonary function.
OUTCOME MEASURES: Outcome measures were sitting posture (referral to a
specialized sitting clinic) and respiratory function (requirement for nocturnal
ventilation) at 17 years of age.
RESULTS: The conservatively managed group included nine patients who had
severe scoliosis at the age of 17 years. The maximum annual Cobb-angle
deterioration was less than 20[degrees] in eight of these nine cases and tended
to occur after 14 years of age. By contrast, the maximum annual Cobb-angle
deterioration among surgical candidates tended to be greater than 20[degrees]
and to occur before the boys were 14 years old. At 17 years of age, patients in
the conservatively managed group tended to have a better sitting posture than
candidates for surgery, as indicated by the proportion of patients who were
referred to a sitting clinic-19% of patients considered not to require surgery
were referred, compared with 33% of patients who received surgery, 36% of
patients who refused surgery and 44% of patients who were unfit for surgery.
Similarly, patients in the conservatively managed group tended to have higher
FVC values and were less likely to develop nocturnal hypoventilation than
surgical candidates (incidence of noninvasive ventilation: 19% among patients
who were considered not to require surgery, 40% among patients who received
surgery, 36% among patients who refused surgery and 56% among patients who were
unfit for surgery).
CONCLUSION: A substantial proportion of DMD patients with scoliosis can
be managed effectively without surgery.
13 - Recommendations to define exercise prescription for duchenne muscular dystrophy
CONCLUSION
Based on the current literature, a major limitation to the use of exercise to treat neuromuscular diseases in general, and DMD specifically, is the absence of systematic analysis to determine the functional capabilities of the diseased muscles. Thus, there is a huge need for conducting studies to assess the benefits of exercise on DMD that range from training studies to determining physiologic outcomes to exploring the molecular signals that define the adaptive responses to the exercise stress. Because of the ethical and practical issues of determining exercise prescription for individuals with DMD, it is suggested that studies may be best conducted first in dystrophic animals (e.g., mice) specifically to determine functional thresholds to maximize the benefits of the exercise and minimize the potential for exacerbating the dystrophy (Fig. 5). However, to effectively transfer these exercise principles to individuals with DMD, noninvasive assays such as those that have been developed for mouse skeletal muscle (1), should be developed for humans to assess beneficial muscle adaptations and potential muscle damage in vivo. Finally, it is suggested that a systematic approach by muscle and cardiovascular physiologists to define exercise prescription should be undertaken in an attempt to positively affect the dystrophic outcome.
13 - MRI for the demonstration of subclinical muscle involvement in muscular dystrophy
13 - A Method to Evaluate Contractures Effects during the Gait of Children with Duchenne Dystrophy
13 - Summary of Important Research Results Between July and December 2006 - Guenter Scheuerbrandt report in English - in Spanish - in Germany
13 - PTC Non-Confidential Report