News in Muscular Dystrophy

NEWS - JANUARY - DECEMBER 2002

December

30/12/02: Important research from 2001 that we don't know nothing more:

CYTRX Announces research collaboration with the University of Cincinnati Medical Center to Study CYTRX's experimental drug (CRL-5861) for treatment of Muscular Dystrophy

Inotrophic effects of the K(+) channel blocker TEA on dystrophic (mdx and dy/dy) mouse diaphragm

Intraarterial Injection of Muscle-derived CD34⁺Sca-1⁺ Stem Cells Restores Dystrophin in mdx Mice

Transfer of full-length Dmd to the diaphragm muscle of Dmd(mdx/mdx) mice through systemic administration of plasmid DNA

Antisense-induced exon skipping restores dystrophin expression in DMD patient derived muscle cells

36 BOYS NEEDED FOR EXPANDED STUDY OF GENTAMICIN IN DUCHENNE MD

Electrotransfer of naked DNA in the skeletal muscles of animal models of muscular dystrophies

Oligonucleotides against a splicing enhancer sequence led to dystrophin production in muscle cells from a Duchenne muscular dystrophy patient

30/12/02: Information from Stephen Kaufman on the Integrin-research (28/12/02):

"Dear Berit,

In the article you refer to we report an interesting and encouraging alternative approach to DMD. We continue to work on this and I believe other labs may also have taken this cue. At this point, unfortunately, we are years away from knowing whether this approach will lead to a meaningful clinical intervention in humans. I assure you we continue to work on this diligently.

Sincerely,

Stephen Kaufman"

28/12/02: Why the 2001 breakthought haven't continuation? In march,2001 I include this news in my brazilian website. After 21 months there are no continuation.

http://www.jcb.org/cgi/content/abstract/152/6/1207

The video (11,5 MB) of this article: http://www.jcb.org/cgi/content/full/152/6/1207/DC1/1 (you need quick time player)

21/12/02: Brazilian University will start in 2003 the study of gene therapy in a dog model of muscular dystrophy

Veterinary Faculty of Universidade de São Paulo (Brazil) inaugurate in this morning the kennel to muscular dystrophy labrador retriever. The cientists of the University will study gene therapy in this dogs using stem cells from blood cord, bone marrow, placenta. This studies will be realized in São Paulo and Ribeirão Preto (Terapy Genic Center).

21/12/02: Brain in DMD and Neuromuscular Diseases:

Diversity of the Brain Dystrophin-Glycoprotein Complex

When Neuromuscular Disease affects the Brain

21/12/02:Spinal fusion and instrumentation for paediatric neuromuscular scoliosis: retrospective review

21/12/02 IN PRESS (International Journal of Cardiology): Assessment of left ventricular systolic and diastolic functions in children with merosin-positive congenital muscular dystrophy

Naci Ceviz, Fusun Alehan, Dursun Alehan, Sencan Ozme, Zuhal Akçoren, Gulsev Kale, Haluk Topaloglu

Abstract
Cardiopathy is an expected finding in X-linked Duchenne and Becker muscular dystrophies. This holds true for some other forms such as autosomal recessive limb-girdle dystrophies. However, data on early-onset and usually severe congenital muscular dystrophies are limited. The purpose of this study was to investigate the presence of cardiac involvement in children with merosin-positive congenital muscular dystrophy. A total of 42 patients and 22 healthy subjects were evaluated by M-mode, 2D, and Doppler echocardiography. Cardiac anatomy, left ventricular dimensions, wall thickness and systolic and diastolic functions were investigated in patients and
compared with those of healthy control subjects. Mean left ventricular ejection fraction and shortening fraction were significantly lower in the patient group (P<0.05 and P<0.001, respectively) and in three patients ejection fraction was below 55%. Although some impairments in left ventricular inflow indexes which were suggestive of left ventricular diastolic dysfunction were detected in patients with merosin-positive congenital muscular dystrophy they were not statistically significant. Our results suggest that left ventricular systolic abnormalities may occur in children with merosin-positive congenital muscular dystrophy.

19/12/02: Enhanced Dystrophic Progression in mdx Mice by Exercise and Beneficial Effects of Taurine and Insulin-Like Growth Factor-1

19/12/02: Researchers at ViaCell Report Additional Successes in Expanding And Preserving Stem Cells from Umbilical Cord Blood for Use in Cellular Therapies

BOSTON, Dec. 18 /PRNewswire/ -- ViaCell, Inc., a premier cellular therapy company, has presented study results demonstrating its ability to expand and preserve purified stem cell populations from umbilical cord blood for future potential use as cellular medicines. Scientists from ViaCell presented data on these as well as other research and development advancements at the recent American Society of Hematology's 44th Annual Meeting in Philadelphia, PA. Among the findings, scientists showed that populations of human umbilical cord blood stem cells that are expanded using ViaCell's patented Selective Amplification(TM) technology can be successfully cryopreserved without any significant loss of function. In a mouse model, these cells were shown to engraft successfully in transplant procedures at a rate that is comparable to that achieved by selectively amplified cells that were not frozen. Study findings indicate that these stem cells can be manufactured and cryogenically frozen in significantly large populations for future use as cellular medicines. "Umbilical cord blood stem cells are used as a part of the therapy regimen for nearly 50 diseases today. One of the challenges in developing additional cellular therapies is the need to multiply and preserve large quantities of these powerful umbilical cord blood stem cells for use in treating an even broader range of diseases. These important studies indicate that we can substantially increase the number of these valuable cells and freeze them for later use," said Jan Visser, Ph.D., senior vice president, head of science at ViaCell. Scientists also reported the ability to control various expression patterns while reproducing a targeted population of umbilical cord blood stem cells using Selective Amplification(TM). The ability to monitor specific cell receptors and ligands offers the potential to engineer specific, homogeneous and high-potency pools of hematopoietic stem cell candidates in large quantities. Data also presented show that the presence of the growth factor thrombopoietin (TPO) in combination with other agents significantly increases the generation of both long- and short-term repopulating stem cells. These findings are now being applied in ViaCell's Selective Amplification(TM) process to expand target populations from various stem cell sources such as bone marrow or cord blood. This is expected to result in the increased production of human hematopoietic stem cell candidates. "We anticipate that these new studies will show that TPO can further enhance production of human hematopoietic stem cells for use in transplantation therapy," added Dr. Visser. Additionally, research progress on the effects of insulin-like growth factor binding protein-3 (IGFBP-3) in modulating the proliferation of primitive stem cells indicates that IGFBP-3 also serves an important function in blocking cell differentiation. The ability to prevent stem cells from differentiating into other cell types could enhance the ability to grow highly enriched populations of stem cells with broad applications in human health. ViaCell first announced the discovery of this protein's impact on regulating the expansion and engraftment potential of human stem cell populations at the 2001 American Society of Hematology's Annual Meeting. "These critically important studies significantly expand ViaCell's history of research discoveries and advances in support of our mission to develop and provide the highest quality cellular therapies for the treatment of many challenging human diseases," said Marc Beer, president and CEO of ViaCell. "The progress reported here contributes significantly to the rapidly evolving field of cellular medicine and therapeutic development." "Our research and development strides presented at this year's American Society of Hematology meeting represent a few of the important accomplishments the ViaCell team has achieved in furthering cellular therapy," said Dr. Visser. "Our progress in areas including other growth factors, antibodies, endogenous stem cell stimulants, pancreatic stem cell and pancreatic islet isolation and expansion methodologies underscores our commitment to the development of innovative cellular therapies to tackle numerous unmet clinical challenges."

16/12/02 IN PRESS (NEUROLOGY 2002;59:1876–1880): Modafinil reduces excessive somnolence and enhances mood in patients with myotonic dystrophy

J.R. MacDonald, PhD; J.D. Hill, MSc; and M.A. Tarnopolsky, MD, PhD

Abstract—Objective: To evaluate the potential of modafinil in reducing excessive daytime somnolence (EDS) and enhancing indexes of quality of life and mood in patients with myotonic dystrophy (DM). Methods: Forty patients with DM were randomized to receive modafinil and placebo for 14 days each, using a double-blind, cross-over design. Before and after each trial, subjects completed handgrip strength testing, spirometry, and quality-of-life measures (RAND). On days 7 and 14, each subject completed the Epworth Sleepiness Scale (ESS), the Stanford Sleepiness Scale (SSS), and the Profile of Mood States (POMS). Results: ESS scores were lower while taking modafinil (mean 248 mm; 95% confidence limit 220 to 276 mm) as compared with placebo (309 mm; 281 to 336 mm) (p < 0.001). Mean SSS scores were also lower during the modafinil trial (3.05; 2.77 to 3.33) than during the placebo trial (3.45; 3.18 to 3.71) (p < 0.05). The POMS indicated that modafinil decreased fatigue–inertia (p < 0.001) and increased vigor–activity and tension–anxiety (p < 0.001) indexes. The total mood disturbance score was also decreased during the modafinil trial as compared with placebo (p < 0.05). TheRAND quality-of-life measures of energy (p < 0.001) and health change (p < 0.05) were both significantly enhanced during the modafinil treatment phase. No changes in maximal grip strength or forced expired volume in 1 second were detected over the course of the study. Headache was the most frequently reported adverse event. Four patients withdrew from the study, three because of side effects (two during modafinil ingestion and one during placebo ingestion). Conclusion: Modafinil reduces somnolence and improves mood in patients with DM.

16/12/02: Novel Gene Therapy Approach Shows Promise

14/12/02 The non-classical MHC molecule HLA-G protects human muscle cells from immune-mediated lysis: implications for myoblast transplantation and gene therapy

14/12/02 Selected abstracts from de 42th Annual Meeting of American Society for Cell Biology (San Francisco, Ca - December, 14-18,2002):

I) TGF-beta triggers the differentiation of myogenic cells toward myofibroblasts: Implication for muscle fibrosis

Y. Li, W. Foster, Y. Chan, B. M. Deasy, T. Payne, T. Horaguchi, N. Badlani, J. Cummins, J. Huard; Dept. of Orthopeadic Surgery, Children's Hospital of Pittsburgh, Pittsburgh, PA

Injured muscle can undergo regeneration; however, the healing process is often inefficient and hindered by progressive fibrosis, which is caused by stimulation of inflammatory factors. Transforming Growth Factor-beta1 (TGF-b1) is considered to be a central factor in fibrotic diseases of the liver, lung, heart, and kidney. Our previous research has found that muscle-derived stem cells are capable of differentiating into myofibroblasts following muscle laceration injury. We also have observed TGF-b1 over-expression in injured skeletal muscle. In this study, we determined that TGF-b1 stimulation can induce myoblasts (C2C12 cells) to express TGF-b1 in an autocrine manner, down-regulate the expression of myogenic proteins, and activate the production of fibrotic-related proteins in vitro. In addition, direct injection of human recombinant TGF-b1 into skeletal muscle in vivo was found to stimulate myogenic cells, including myofibers, to express TGF-b1 and subsequently induce scar tissue formation within the injected area. These results demonstrate that TGF-b1 is a key factor that can initiate the fibrotic cascades in skeletal muscle and induce a population of myogenic cells, which includes myofibers, to de-differentiate into myofibroblastic cells in injured muscle. These observations shed new light on the process of scar tissue formation, which commonly occurs in injured and diseased muscle.

II) Side population stem cells are recruited to sites of acute but not chronic muscle degeneration

K. A. Lapidos, E. M. McNally; Cardiology, University of Chicago, Chicago, IL

Muscular dystrophies are degenerative disorders in which skeletal muscle undergoes chronic cycles of degeneration and regeneration. Stem cells are a critical component of the regenerative process. Recently, a pluripotent population of stem cells has been isolated from bone marrow and muscle of adult mice, and these side population (SP) cells were shown to regenerate hematopoietic lineages and skeletal muscle. Based on their ability to differentiate into both cell types, we hypothesized that SP cells comprise an in vivo, stable stem cell compartment available for the regeneration of muscle tissue or the hematopoietic lineages. For example, in muscular dystrophy, a high demand for stem cells in the muscle promotes recruitment of SPs from the bone marrow. Over time, this may result in a quantifiable decrease in SP cells from bone marrow and a quantifiable, concomitant increase at the muscle. We tested this hypothesis by quantifying the percentage of SP cells obtained from bone marrow or muscle in a chronic model of muscular dystrophy, g-sarcoglycan null mice (gsg^-/-). Whole bone marrow and lower limb skeletal muscle were isolated from wild type and gsg^-/- mice at either 6 weeks or 6 months of age. Using the Hoechst dye exclusion technique, SPs from bone marrow (HSPs) and muscle (MSPs) were quantified. We found no statistically significant differences between the mutant and wild type mice. We also studied an acute model of muscle degeneration elicited by cardiotoxin injection of the crural muscles of normal mice. Three days post-injection, SP cells showed a five to nine-fold increase in MSPs in cardiotoxin-treated versus the control. In conclusion, we have shown that side population cells increase at the site of acute, but not chronic, muscle degeneration/regeneration.

III) Staging Interactive Pathogenic Mechanisms in mdx Mouse Dystrophinopathy by Temporal Gene Expression Signatures

J. D. Porter, A. P. Merriam, S. Khanna; Ophthalmology, Case Western Reserve University, Cleveland, OH

Although Duchenne muscular dystrophy (DMD) results from mutations of dystrophin, multiple interactive mechanisms likely contribute toward pathogenesis. We used Affymetrix oligonucleotide arrays to compare hindlimb muscle of the mdx mouse dystrophinopathy to wild type mice at postnatal (P) ages 7, 14, 18, 23, 28, 56, and 112 days; 3 independent replicates/age/strain. Extraocular muscle, which shows no signs of pathology in dystrophinopathy, also was studied at most ages. Patterned alterations in hindlimb gene expression between P7 and P14 (prenecrotic stage) included only 8 genes. Rapid accumulation of expression changes was observed thereafter, with a 20-fold increase in the number of up-regulated genes between P14 and P28, which coincided with appearance of histologic changes in muscle. Assessed at the expression level, disease peaked with 376 differentially regulated genes at P56, but a persistent response was evident at P112 (~130 genes). Correlation analyses provided insight into the interactive nature of inflammatory, fibrotic, and muscle regenerative events, with proinflammatory mechanisms particularly evident. By contrast, few changes were detected in extraocular muscle at any age. Immunocytochemistry and quantitative PCR for chemotaxic molecules confirmed their early role in recruitment of an inflammatory infiltrate to dystrophic muscle, along with a corresponding absence in extraocular muscle. These findings suggest that inflammation is a key contributor to, rather than a consequence of, the pathogenesis of dystrophin deficiency. The absence of changes in extraocular muscle suggest that this tissue does not adapt, but rather is constitutively protected in muscular dystrophy. Taken together, genome-wide expression profiling establishes an integrated profile of dystrophinopathy and the resultant mdx temporal expression signature for hindlimb suggests that blockade of inflammatory cell recruitment may be effective in ameliorating the disease, an attractive prospect in the absence of effective therapy for the primary mutation.

IV) Migration and Fusion of Muscle SP Cells into Dystrophic Myofibers

R. L. Sohn,¹ E. Gussoni² ; ¹ Medicine/Cardiology, Brigham and Women's Hospital, Boston, MA, ² Division of Genetics, Children's Hospital, Boston, MA

Cell therapy with muscle SP cells in animal models is being explored as a potential treatment for muscular dystrophy. However, though promising, stem cell therapy is plagued by its low efficiency: only about 1-5% of these stem cells fuse into pre-existing myotubes. In fact, how myoblasts, or even muscle SP cells, fuse to form multinucleate myotubes is a fascinating and highly regulated process. Recent work in Drosophila has elucidated some of the molecular components of this process. Our hope is to improve the efficiency of cell therapy with muscle SP cells by utilizing the tools and concepts of myoblast fusion found in Drosophila. As a first step, we have looked for cell surface markers that promote the migration of SP cells from the blood where they are injected to skeletal muscle where they then fuse into myotubes. Chemokines are known to play major roles in leukocyte migration. Therefore, we looked for the presence of chemokine receptors in skeletal muscle. By immunohistochemistry on human fetal skeletal muscle, we have found cells positive for the chemokine receptor CXCR-4 along the myofibers. These are not satellite cells as they are not positive for the transcription factor Pax-7. We have also found myotubes positive for CDO (CAM-regulated, down-regulated by oncogenes), a cell surface protein of the immunoglobulin/fibronectin type III repeat family that has been found to positively regulate myogenic differentiation in vitro (Kang, et al. 1998, JCB 143: 403-13). Work is currently underway to determine if these cell surface proteins are also present in muscle SP cells. If so, it is possible that by increasing the expression of these molecules, we will be able to also increase the efficiency of the SP cells to migrate to muscle and to fuse into the myotubes.

14/12/02: Cloning embryos a ‘pro-life’ move

14/12/02: U. of Minnesota grants rights to develop adult stem-cell technology

14/12/02: Temporomandibular joint and masticatory muscle involvement in myotonic dystrophy: a study by magnetic resonance imaging

9/12/02: New information about the Transgene research obtained by our friend from Norway, Berit Sofie: "Current protocol of a research phase I clinical trial of full-length dystrophin plasmid DNA in Duchenne/Becker muscular dystrophies Part II: clinical protocol"

e-mail from Serge Braun, PharmD, PhD - Vice-President Research Transgene S.A.:

Dear Berit,

We are working on gene therapy of Duchenne dystrophy, but still at a preliminary stage (phase I clinical trial). As you may have heard, we are currently completing a phase I clinical trial with a dystrophin-plasmid non-viral vector injected intra-muscularly. The goal is to demonstrate good tolerance and safety of the vector and of the newly produced dystrophin. The next step is to take the intra-arterial delivery procedure to the clinic (trial expected 2004); at this stage we would like to target a small foot muscle, again for safety reasons. Should it work, we would than move on to a more ambitious treatment, getting to whole limb muscles (expected 2006 ?). Then, we would target respiratory and cardiac muscles. But, there is still a long way to go.

I understand time is running too fast for the Duchenne kids and families. It is however impossible to proceed faster for security reasons. It would be a catastrophy should a severe side effect or any undesired event happend.

Keep in touch.

Serge Braun

7/12/02: Prevention of pathology in mdx mice by expression of utrophin: analysis using an inducible transgenic expression system

7/12/02: Effects of corticosteroids in the management of Duchenne muscular dystrophy: our experience

Full text in Spanish: Efecto de los corticoides en el tratamiento de la distrofia muscular de Duchenne

5/12/02: Improved Contractile Function of the mdx Dystrophic Mouse Diaphragm Muscle after Insulin-Like Growth Factor-I Administration

5/12/02 IN PRESS (NEUROMUSCULAR DISORDERS): High dose weekly oral prednisone improves strength in boys with Duchenne muscular dystrophy

Anne M. Connolly, Jeanine Schierbecker, Renee Renna, Julaine Florencea

Department of Neurology, Saint Louis Children’s Hospital, Washington University School of Medicine, Box 8111, 660 S. Euclid Avenue,63110 Saint Louis, MO, USA

Department of Pediatrics, Saint Louis Children’s Hospital, Washington University School of Medicine, Box 8111, 660 S. Euclid Avenue,63110 Saint Louis, MO, USA

Abstract

Daily prednisone improves strength in boys with Duchenne muscular dystrophy, but side effects are almost universal. We used a different dosing regimen of prednisone to determine if benefit to boys with Duchenne muscular dystrophy might be maintained with fewer side effects. Twice weekly oral prednisone was given each Friday and Saturday (5 mg/kg/dose). This total dose is twice as high as the daily low dosage prednisone regimen (0.75 mg/kg/day). Twenty boys (8.0 ^ 1.2 years) were treated. Historical control groups included 18 untreated boys (6.1 ^ 1.6 years) and four boys (7.3 ^ 0.6 years) treated with daily prednisone. Strength (using a hand-held manometer and grip meter) and timed functional testing were measured. There was an improvement in upper extremity strength for 95% of boys (n ¼ 20) at 6 months using quantitative strength testing. Improvement in lower extremity strength occurred in all boys with antigravity quadriceps strength (17/17). The improvement (P ¼ 0:001 for proximal upper extremities; P ¼ 0:002 for grip; and P , 0:0001 for proximal lower extremities) was significant compared to untreated boys. Sixteen boys were treated continuously for more than 12 months (22 ^ 1.5 months). Of these, 15 remained significantly stronger than prior to treatment and 8/16 showed additional gains in strength after six months of treatment. Six boys were on the weekly prednisolone 2 years or longer without interruption. All six had upper and lower extremity strength at follow-up that was as good or better than at baseline. Functional testing improved in boys less than 8 years without contractures. Three boys without antigravity quadriceps strength at the start of treatment lost the ability to walk unassisted within 6 months. Eight other boys lost the ability to ambulate unassisted between 12 and 24 months of treatment. In each, progressive contractures developed. Linear growth was maintained in all boys on weekly treatment. Obesity rates did not differ from untreated boys. Twice weekly prednisone improved strength over 6–12 months in the majority of boys, but did not slow contracture development. Sustained benefit beyond 12 months is possible with fewer side effects compared to daily prednisone.

5/12/02: Australia OKs embryo stem cell research

November

30/11/02: Results of this research "Current protocol of a research phase I clinical trial of full-length dystrophin plasmid DNA in Duchenne/Becker muscular dystrophies Part II: clinical protocol" will be published only in 2003.

I receive this e-mail from Institute de Myologie, France about this research:

"Dear Dr. Feder,

Thank you very much for your mail and for your interest in neuromuscular
disorders. To answer your question, the trial on Duchenne muscular
dystrophy is not yet completely finished, currently the last patient is
enrolled. This means that the results of the trial will probably be known
by early next year.

I hope this is helpful,
Yours sincerely,

Jacques Salama
Secretaire General"

The summary of this article: Current protocol of a research phase I clinical trial of full-length

dystrophin plasmid DNA in Duchenne/Becker muscular dystrophies Part II: clinical protocol

Norma Beatriz Romeroa,*, Olivier Benvenisteb, Christine Payana, Serge Braunc, Patrick Squibanc,Serge Hersonb, Michel Fardeaua

A phase I clinical study on gene therapy in Duchenne and Becker muscular dystrophy, open, without direct individual benefit for thepatient, is being performed at the Pitie´-Salpeˆtrie`re Hospital, Paris. The aims of this project are: (a) to determine the tolerance and the safety of the intramuscular administration of dystrophin cDNA and (b) to study the quality of the gene transfer in vivo in human patients affected byDuchenne and Becker muscular dystrophy. This clinical trial is conducted sequentially and includes three cohorts of three patients each. Patients must be at least 15 years of age. Diagnosis of Duchenne and Becker muscular dystrophy was confirmed by molecular analysis of the dystrophin gene and for each patient the abnormal expression of dystrophin was confirmed, in skeletal muscle, with antibodies directed against the deleted part of the dystrophin. This phase I study is scheduled to be completed by the end of 2002.

30/11/02: Defective glycosylation in muscular dystrophy

Francesco Muntoni (e-mail:[email protected]), Martin Brockington, Derek J Blake, Silvia Torelli, Susan C Brown Lancet 2002; 360: 1419-21

Context Over the past 15 years the causative genes of several inherited muscular dystrophies have been identified. These genes encode sarcolemmal, extracellular matrix, sarcomeric, and nuclear envelope proteins. Although the post-translational processing of muscle proteins has a significant role in their correct assembly and function, these processes have not been shown to be primarily involved in the pathogenesis of muscular dystrophies until recently. In the past 18 months, four different forms of inherited muscular dystrophy in human beings have been associated with mutations in genes encoding for putative glycosyltransferases. Aberrant glycosylation of -dystroglycan, an external membrane protein expressed in muscle, brain, and other tissues, is a common feature in these disorders. -dystroglycan is highly glycosylated, its sugar components varying in different tissues and controlling its interaction with extracellular matrix partners. Disrupted glycosylation of -dystroglycan results in a loss of these interactions, giving rise to both progressive muscle degeneration and abnormal neuronal migration in the brain.

Starting point Kevin Campbell and colleagues have recently demonstrated that patients with muscle-eye-brain disease (MEB) and Fukuyama congenital muscular dystrophy (FCMD), as well as the myodystrophy (myd) mouse, have an abnormally glycosyated form of -dystroglycan (Nature 2002; 418: 417-22 and 422-25). The abnormally glycosylated protein did not bind to three of its extracellular matrix ligands, laminin 2 chain, agrin, and neurexin. The investigators also showed that a neuronal migration disorder occurs in both the myd mouse and in a brain-restricted -dystroglycan knock-out mouse that is similar to that seen in patients with MEB and FCMD. These results identify -dystroglycan as having an essential role in both muscle and brain development and function.

Where next Emphasis is moving away from identifying the protein components of the muscle fibre that are involved in muscular dystrophies towards the post-translational processing of proteins and the enzymes involved in these modifications. This opens up new avenues of research. Abnormal glycosylation of -dystroglycan may underlie other as yet uncharacterised forms of muscular dystrophy and neuronal migration disorders.

30/11/02: Tailor-made sugar coated proteins manufactured in novel E. coli system

28/11/02: Muscular dystrophy: Myostatin blockade

Myostatin is a negative regulator of muscle mass — a mutant myostatin is responsible for the exaggerated musculature in Belgian blue cattle. Blockade of myostatin activity in the mdx mouse model for Duchenne muscular dystrophy with a monoclonal antibody is now shown to improve muscle function and cause a significant increase in muscle size. Myostatin blockade would be an attractive strategy for treating muscle degeneration diseases, as it would avoid some of the problems associated with conventional gene therapy.

Functional improvement of dystrophic muscle by myostatin blockade

Nature 420, 418 - 421 (2002); doi:10.1038/nature01154

SASHA BOGDANOVICH*†, THOMAS O. B. KRAG*†, ELISABETH R. BARTON*, LINDA D. MORRIS*, LISA-ANNE WHITTEMORE‡, REXFORD S. AHIMA§ & TEJVIR S. KHURANA*

* Department of Physiology and Pennsylvania Muscle Institute, University of Pennsylvania School of Medicine, 3700 Hamilton Walk, Richards A-601, Philadelphia, Pennsylvania 19104-6085, USA
‡ Musculoskeletal Sciences Department, Wyeth Research, Cambridge, Massachusetts 02140, USA
§ Division of Endocrinology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA
† These authors contributed equally to this work

Correspondence and requests for materials should be addressed to T.S.K. (e-mail: [email protected]).

Mice and cattle with mutations in the myostatin (GDF8) gene show a marked increase in body weight and muscle mass, indicating that this new member of the TGF- superfamily is a negative regulator of skeletal muscle growth. Inhibition of the myostatin gene product is predicted to increase muscle mass and improve the disease phenotype in a variety of primary and secondary myopathies. We tested the ability of inhibition of myostatin in vivo to ameliorate the dystrophic phenotype in the mdx mouse model of Duchenne muscular dystrophy (DMD). Blockade of endogenous myostatin by using intraperitoneal injections of blocking antibodies for three months resulted in an increase in body weight, muscle mass, muscle size and absolute muscle strength in mdx mouse muscle along with a significant decrease in muscle degeneration and concentrations of serum creatine kinase. The functional improvement of dystrophic muscle by myostatin blockade provides a novel, pharmacological strategy for treatment of diseases associated with muscle wasting such as DMD, and circumvents the major problems associated with conventional gene therapy in these disorders.

28/11/02: Blocking Myostatin Proves Beneficial in Mice with DMD

25/11/02: Loss of myostatin attenuates severity of muscular dystrophy in mdx mice

Johns Hopkins release

Comments:1) Mighty mice are less susceptible to muscular dystrophy gene's effects

2) Mighty Mice Are Less Susceptible To Muscular Dystrophy Gene's Effects

23/11/02 IN PRESS (NEUROMUSCULAR DISORDERS): Report on the Muscular Dystrophy Campaign workshop: Exercise in neuromuscular diseases

Summary: It must be emphasised that these recommendations are based on current expert opinion only and that research is needed to improve the evidence base in all the areas suggested.

A. Ambulant children

1. Daily stretches to the gastrocnemius–soleus complex, hip flexors and iliotibial band.

2. Encourage voluntary active exercise such as swimming or hydrotherapy and cycling (may be motor assisted).

3. Symmetry to be promoted in posture, exercises and activities.

4. Eccentric activities such as running downhill and excessive walking downstairs to be avoided.

B. Non-ambulant children

1. Mobilising passive or active assisted exercises to maintain and promote symmetry and comfort. These may be land based programmes or in water if preferred.

Wheelchair/seating prescription, respiratory and spinal management have not been discussed in this workshop but they are clearly important issues that need consideration in the holistic management of the young person.

C. Ortosis

1. Night-time AFOs in addition to stretching daily are recommended for ambulant children with DMD to maintain the length of the gastrocnemius–soleus complex. There is no evidence on when to supply night splints but it is recommended that this be when there is loss of dorsiflexion.

2. AFOs are not recommended for ambulant children with DMD as this compromises their ability to walk by preventing characteristic equinus gait. In ambulant children with other neuromuscular disorders careful assessment is essential to ensure that walking is not compromised.

3. Clinical experience suggests that daytime AFOs should be supplied once ambulation is lost to prevent painful contractures and foot deformity. If tenotomies are performed in the non-ambulant child AFOs should be worn during the day.

4. KAFOs can be used to prolong ambulation for approximately 2 years in DMD. They can also help delay the onset of lower limb contractures and weaker evidence suggests that prolonging ambulation beyond 13 years may delay the onset of scoliosis. They should be supplied at the time of loss of ambulation by an orthotist with experience in neuromuscular disorders.

5. The benefit of a standing posture in the non-ambulant child to control contractures is logical but not evidenced. Standing frames or swivel walkers may be used in children with neuromuscular disorders.

D. Adults with muscular dystrophy

1. An accurate diagnosis should be made so that the possible complications and manifestations of the disease can be considered in devising a physical treatment plan and assessment programme.

2. The possibility of overuse fatigue, pain or weakness should be considered especially in FSH muscular dystrophy but there is no reliable evidence to suggest that exercise is contraindicated.

3. Active low resistance exercise may be prescribed to improve strength and endurance in relatively well maintained muscles (stronger than grade 3 MRC scale).

4. Where possible active exercise on a regular basis to promote general physical health to be encouraged.

5. Patients with myotonic dystrophy may be given an active resisted exercise regime but may need additional support and motivation.

6. Avoid inspiratory breath holding techniques in myotonic dystrophy.

7. AFOs may be used in ambulant patients with foot drop but not effective if plantar grade cannot be achieved

23/11/02 IN PRESS (NEUROMUSCULAR DISORDERS): Steroids in Duchenne muscular dystrophy: from clinical trials to genomic research Francesco Muntonia,*, Ivan Fishera,b,c, Jennifer E. Morganb, David Abrahamc

Abstract

Steroids represent the only pharmacological palliative treatment for Duchenne muscular dystrophy. However, they do have side effects and despite a large number of published studies showing their efficacy, they are still not universally used. This is largely due to the lack of functional outcome and quality of life measures in most of the published studies and suggests that further trials might be required to answer some of the still unclear aspects of their role. Another important aspect of steroid therapy in Duchenne dystrophy is that we do not know how they work in dystrophic muscle. We have initiated a collaborative study on gene profiling using microarray in steroid-treated mdx mice. cDNA microarray studies were performed to examine the levels of skeletal muscle gene expression in a pool of mdx mice treated with prednisolone for 1 and 6 weeks. Interesting preliminary data on untreated mdx mice suggest that the gene profiling of young (7 weeks) versus older (12 weeks) mice is very significantly different. Furthermore, a large number of genes showed significant changes in expression at the mRNA level on treatment with prednisolone. These included structural protein genes; signalling genes and genes involved in immune response. Hopefully, analysis of this pattern of steroid-induced gene expression will provide some insight into understanding how glucocorticoids improve strength in Duchenne dystrophy, and may help in developing more effective and less toxic therapeutic approaches.

23/11/02: Muscle cell transplants repair damaged heart tissue

23/11/02: Gene therapy eases stubborn heart disease

20/11/02: Improved antisense oligonucleotide induced exon skipping in the mdx mouse model of muscular dystrophy.

18/11/02 IN PRESS (NEUROMUSCULAR DISORDERS): An evaluation of leukaemia inhibitory factor as a potential therapeutic agent in the treatment of muscle disease

The exogenous delivery of growth factors and cytokines is a potential therapeutic strategy to alleviate the degenerative effects of primary inherited myopathies such as Duchenne muscular dystrophy. The mdx mouse diaphragm is a model for examining the progressive degeneration of dystrophic muscle. We have delivered leukaemia inhibitory factor to the mdx diaphragm using slow release alginate gels. Previous studies have reported an improvement in the histology of mdx diaphragms after delivery of leukaemia inhibitory factor in a similar manner, but little attention has been paid to the mechanism by which leukaemia inhibitory factor acts. We have used autoradiography to examine cell proliferation, Evans Blue Dye to examine myofibre damage, and morphometric analysis to examine histology in leukaemia-inhibitory-factor-treated diaphragms and compared them with untreated mdx and normal C57Bl10/ScSn diaphragms. Autoradiography showed that although myoblast proliferation was significantly increased in leukaemia inhibitory factor-treated mdx diaphragms, leukaemia inhibitory factor did not reduce myofibre damage and no histological improvement was observed. The data presented here, while demonstrating a role for leukaemia inhibitory factor in myoblast proliferation, do not support a strong and consistent benefit of leukaemia inhibitory factor on dystrophic muscle in vivo as a means of alleviating the effects of chronic dystrophic muscle degeneration.

18/11/02 IN PRESS (NEUROMUSCULAR DISORDERS): Survival in Duchenne muscular dystrophy: improvements in life expectancy since 1967 and the impact of home nocturnal ventilation

We reviewed the notes of 197 patients with Duchenne muscular dystrophy whose treatment was managed at the Newcastle muscle centre from 1967 to 2002, to determine whether survival has improved over the decades and whether the impact of nocturnal ventilation altered the pattern of survival. Patients were grouped according to the decade of death and whether or not they were ventilated. Kaplan Meier survival analyses showed significant decade on decade improvement in survival. Mean age of death in the 1960s was 14.4 years, whereas for those ventilated since 1990 it was 25.3 years. Cardiomyopathy significantly shortened life expectancy from 19 years to a mean age of 16.9 years. Better coordinated care probably improved the chances of survival to 25 years from 0% in the 1960s to 4% in the 1970s and 12% in the 1980s, but the impact of nocturnal ventilation has further improved this chance to 53% for those ventilated since 1990.

16/11/02: Scientists show how stem cells change Stanford uses fluorescent mice to track transformation into tissue

16/11/02: A- and B-utrophin Have Different Expression Patterns and Are Differentially Up-regulated in mdx Muscle

16/11/02: Beta-2-Agonist fenoterol has greater effects on contractile function of rat skeletal muscles than clenbuterol

16/11/02: Long-term follow-up of arrhythmias in patients with myotonic dystrophy treated by pacing

15/11/02: "HOW THE MDA CLINIC CAN MAKE A DIFFERENCE" IS FOCUS OF MDA NATIONAL CONFERENCE

13/11/02: MDA PHYSICIANS REVIEW PROGRESS IN FOUR DISEASES

13/11/02: Immunological hurdles in the path to gene therapy for Duchenne muscular dystrophy

13/11/02: Predictive Factors of Cessation of Ambulation in Patients with Duchenne Muscular Dystrophy

Bakker JPJ, de Groot IJM, Beelen A, Lankhorst GJ: Predictive factors of cessation of ambulation in patients with Duchenne muscular dystrophy.

Am J Phys Med Rehabil 2002;81:906–912.

Objectives: To identify baseline patient and treatment characteristics that can predict wheelchair dependency within 2 yr.

Design: This prospective cohort study included 44 subjects who met study inclusion criteria. The same investigator examined them at 6-mo intervals. Ambulatory status, anthropometric data, muscle strength, range of motion of weight bearing joints, scoliosis, WeeFIM® instrument, Functional Status II revised, and use of standing and walking aids. Cox proportional hazards regression analysis and the stepwise technique were used to search for prognostic factors of wheelchair dependency within 2 yr.

Results: Children with impaired hip extension and ankle dorsiflexion strength are 11.5 (95% confidence interval, 3.2– 40.5) and 3.7 (95% confidence interval, 1.4 –9.7) times, respectively, more likely to stop ambulating within 2 yr.

Conclusions: This study confirms that strength loss, specifically in hip extension and ankle dorsiflexion, are the primary predictors of loss of ambulation in Duchenne muscular dystrophy. Further research is needed for medical interventions that can improve hip extension or ankle dorsiflexion and actually can improve ambulation.

09/11/02: Prenatal stem cell transplants may open door to organ transplants, treating genetic diseases

09/11/02: Adeno-associated virus vector-mediated gene transfer into dystrophin-deficient skeletal muscles evokes enhanced immune response against the transgene product

09/11/02: Fracture prevalence in Duchenne muscular dystrophy.

04/11/02: ADAM12 Alleviates the Skeletal Muscle Pathology in mdx Dystrophic Mice

02/11/02: Gene expression comparison of biopsies from Duchenne muscular dystrophy (DMD) and normal skeletal muscle

02/11/02: The role of CD34 expression and cellular fusion in the regeneration capacity of myogenic progenitor cells

02/11/02: Men With Duchenne MD Find Independence Within Their Reach

02/11/02: Loss of sarcolemma nNOS in sarcoglycan-deficient muscle.

02/11/02: Fibronectin receptor reduction in skin and fibroblasts of patients with Ullrich's disease.

OCTOBER:

26/10/02: ABSTRACTS - THE ANNUAL METTING OF AMERICAN SOCIETY OF HUMAN GENETICS

26/10/02: Fibrogenic cytokines and extent of fibrosis in muscle of dogs with X-linked golden retriever muscular dystrophy.

26/10/02: Stem cells from cord blood show promise

21/10/02: UI muscular dystrophy findings may point the way to muscle regeneration therapies

19/10/02: Excellent news for Duchenne Muscular Dystrophy: A promising treatment tested in Quebec City

http://montreal.cbc.ca/template/servlet/View?filename=muscdist021016

http://www.cbc.ca/stories/2002/10/17/musc_dys021017

19/10/02: New drugs from nonsense

12/10/02: Improved Performance of a Fully Gutted Adenovirus Vector Containing Two Full-Length Dystrophin cDNAs Regulated by a Strong Promoter

12/10/02: FDA panel: ‘Bubble boy disease’ gene therapy trial should go forward

9/10/02: Transgene and Mirus Reach Research Milestone - Application in Intra-arterial Gene Delivery to Muscle Cells

5/10/02:Bubble Boy' Gene Trials Halt When Boy Gets Cancer

SEPTEMBER

28/09/02: Overexpression of a calpastatin transgene in mdx muscle reduces dystrophic pathology

28/09/02: Ullrich disease: Collagen VI deficiency: EM suggests a new basis for muscular weakness

23/09/02: Antibiotic May Help Fight Genetic Diseases

23/09/02: Calif. to Enact Bill Promoting Stem Cell Research

21/09/02: Delivery of alpha- and beta-sarcoglycan by recombinant adeno-associated virus: efficient rescue of muscle, but differential toxicity

19/09/02: MDA - Application of Stem Cell Therapy in Treating Neuromuscular Diseases:

September 13, 2002

TO: Families of Those Affected by Duchenne and Becker Muscular Dystrophy

FROM:     Robert Ross
                President & CEO
                Muscular Dystrophy Association

RE: Application of Stem Cell Therapy in Treating Neuromuscular Diseases

Stem cell research is an emerging area of scientific investigation which holds extraordinary promise for advancing the development of therapies for the muscular dystrophies and other neuromuscular disorders. Media coverage of reports on the use of stem cells has been extensive . A September 10, 2002 article in The New York Times (copy attached) summarizes one such report. The following is offered to help place this and similar reports in a realistic context.

The Times article indicates that MDA Scientific Advisory Committee Member Louis M. Kunkel, Ph.D. and his team have verified the presence of donor cells in the skeletal muscle of a young man with Duchenne muscular dystrophy who had earlier received a bone marrow transplant (a source of stem cells) for an immune deficiency disease. Although it's significant that some donor cells were found in his muscle tissue, the efficiency of the procedure was very low.

While researchers are optimistic that stem cell therapy will be used to effectively treat neuromuscular disorders, the low efficiency of stem cell incorporation into muscle at this point means that the therapeutic application of stem cell technology is still a number of years in the future.

The Association has taken steps to accelerate stem cell research by sponsoring meetings of specialists to review and evaluate progress, and has established a stem cell working group to guide the Association's efforts in this area. At its most recent meeting the group focused on identifying a series of critical milestones that must be met in order to conduct a bone marrow transplant-based clinical trial. At present, the risk/benefit ratio of such a clinical trial in youngsters with neuromuscular disease does not support it.

Information about the status of specific neuromuscular disease clinical trials is available on the Association's Web site at www.mdausa.org by clicking on "Research" and "Active Clinical Trials"."

18/09/02: Stanford researchers devise novel gene therapy technique

18/09/02: Transgene’s Portfolio Expanded by Ten Patents

17/09/02: Gene therapy reverses muscular dystrophy in animal model

14/09/02: Immune Response to Full-Length Dystrophin Delivered to Dmd Muscle by a High-Capacity Adenoviral Vector

14/09/02: Pulsed Doppler tissue imaging in dystrophinopathic cardiomyopathy

07/09/02: Adeno-Associated Virus Vector-Mediated Minidystrophin Gene Therapy Improves Dystrophic Muscle Contractile Function in mdx Mice

07/09/02: Mice deficient in small leucine-rich proteoglycans: novel in vivo models for osteoporosis, osteoarthritis, Ehlers-Danlos syndrome, muscular dystrophy, and corneal diseases

07/09/02: The ß₂-agonist fenoterol has greater effects on contractile function of rat skeletal muscles than clenbuterol

03/09/02: $58.3 MILLION! JERRY LEWIS TELETHON SETS RECORD

AUGUST

24/08/02: Heart drug could help muscle growth

24/08/02: -Sarcoglycan, a novel component of the sarcoglycan complex, is reduced in muscular dystrophy

17/08/02: ViaCell Obtains Exclusive Worldwide License to Novel Protein for Use in Treatment of Stroke Recovery and Other Neurological and Neuromuscular Disorders
Dimerized Fibroblast Growth Factor (dFGF) Shows Potential to Enhance Neurological Recovery After Stroke

BOSTON, Aug. 14 /ViaCell Inc., a premier cellular therapy company, announced that it has obtained an exclusive worldwide license for dimerized fibroblast growth factor (dFGF), a novel protein developed by
scientists at Massachusetts Institute of Technology (M.I.T.), with applications for the treatment of stroke and other neurological disorders.
ViaCell Neuroscience, a subsidiary of ViaCell, will initially develop dFGF as a product to enhance functional recovery in patients who have suffered a stroke.Marc Beer, ViaCell's chairman and chief executive officer, stated, "Our team at ViaCell Neuroscience is committed to the development of new stem cell
and stem cell stimulating therapies to treat neurological diseases. We are very encouraged by the development of dFGF, which represents a major advance in our potential to treat stroke and numerous other neurological disorders."
ViaCell Neuroscience's exclusive license includes rights to intellectualn property covering dFGF and its use in treatment of all neurological disorders, including stroke. These patents are based on inventions of Ram Sasisekharan, Ph.D., professor in the department of bioengineering and environmental health
at M.I.T.
"We believe that this growth factor may prove to be a very important contributor in ongoing research to develop effective therapies to treat neurological diseases," said Dr. Sasisekharan.
Fibroblast growth factors (FGFs) are a family of proteins in the human body responsible for the proliferation and survival of cell types that comprise the bulk of living cells in many tissues, including brain, vascular
system, and muscle. Dimerized fibroblast growth factor (dFGF) is a novel combination of two subunits of FGF protein that have been linked together.
Over the past 15 months, a team of researchers at ViaCell Neuroscience has found that the intravenous administration of dFGF markedly enhances neurological recovery in an animal model of stroke, even when given a day or more after the event has occurred. The effect is most likely due to stimulation of endogenous stem cells within the brain.
"The results to date are very encouraging," said Seth Finklestein, M.D., head of ViaCell Neuroscience and vice president of ViaCell, Inc. "We believe dFGF is an important protein that has the potential to be a breakthrough
therapy for stroke and other neurological disorders."
Development of dFGF therapies will be conducted at the ViaCell research and development facility in Worcester, MA, under the direction of Dr.Finklestein. Prior to joining ViaCell, Dr. Finklestein served as an associate professor of neurology at Harvard Medical School and was also associate neurologist at Massachusetts General Hospital for more than 12 years. He continues his clinical activities at Massachusetts General Hospital.
While the effects of stroke can be devastating, there are no treatments available today to enhance patient recovery. The only approved therapy for treating stroke is tissue plasminogen activator (tPA) which has been shown to be useful in minimizing the initial damage from stroke. However, treatment
must be administered within three hours of stroke onset.
"With dFGF, we have the potential to provide an entirely new therapy for stroke patients, namely the administration of a treatment days or even weeks after a stroke has occurred, in order to improve neurological function and quality of life," said Dr. Finklestein.
Stroke or "brain attack" is a clot (ischemia) or hemorrhage in the brain that frequently results in damage to brain tissue. Stroke is the third leading cause of death in the United States, killing nearly 160,000 Americans
each year. An additional 590,000 Americans experience and survive a new or recurrent stroke each year; two-thirds of these patients live with moderate to severe paralysis, or speech, vision, or memory loss. There are four million stroke survivors in the U.S. The National Stroke Association estimates that stroke costs the United States $30 billion annually.
About ViaCell, Inc.
ViaCell, Inc. (http://www.viacellinc.com) is a clinical-stage biotechnology company with a cellular medicine pipeline that focuses in the areas of cancer, neurological diseases, diabetes and muscular dystrophy.
ViaCell offers umbilical cord blood stem cell preservation services through Viacord (http://www.viacord.com), a subsidiary of ViaCell. ViaCell is developing amplified, high-definition stem cell products based on its patented technology, Selective Amplification(TM), for the expansion of targeted populations of stem cells. ViaCell is currently conducting a Phase I clinical trial with its lead product, produced through Selective Amplification(TM). ViaCell Neuroscience, a subsidiary of ViaCell, is committed to the development
of new molecular and cellular treatments to prolong life and restore function in neurological diseases, including stroke, head trauma, amyotrophic lateral sclerosis (ALS), genetic enzyme deficiencies and muscular dystrophy, among others.

17/08/02: Patterns and predictors of sleep disordered breathing in primary myopathies.

17/08/02: Tumor necrosis factor-alpha and myocardial function in patients with myotonic dystrophy type 1

17/08/02: Patients + research = result! The role of patients and their interest groups in biomedical research

17/08/02: The therapeutic potential of stem cells from adults

10/08/02: Duchenne muscular dystrophy: current knowledge, treatment, and future prospects

10/08/02: Human feeders support prolonged undifferentiated growth of human inner cell masses and embryonic stem cells

Mark Richards¹, Chui-Yee Fong¹, Woon-Khiong Chan², Peng-Cheang Wong¹ & Ariff Bongso¹
Correspondence should be addressed to A Bongso. e-mail: [email protected]

Previous reports have demonstrated the growth of undifferentiated human embryonic stem (HES) cells on mouse embryonic fibroblast (MEF) feeders and on laminin- or Matrigel-coated plastic surfaces supplemented with MEF-conditioned medium^1-3. These xenosupport systems run the risk of cross-transfer of animal pathogens from the animal feeder, matrix, or conditioned medium to the HES cells, thus compromising later clinical application. Here we show that human fetal and adult fibroblast feeders support prolonged undifferentiated HES cell growth of existing cell lines and are superior to cell-free matrices (collagen I, human extracellular matrix, Matrigel, and laminin) supplemented with human or MEF feeder–conditioned medium. Additionally, we report the derivation and establishment of a new HES cell line in completely animal-free conditions. Like HES cells cultured on MEF feeders, the HES cells grown on human feeders had normal karyotypes, tested positive for alkaline phosphatase activity, expressed Oct-4 and cell surface markers including SSEA-3, SSEA-4, Tra 1-60, and GCTM-2, formed teratomas in severely combined immunodeficient (SCID) mice, and retained all key morphological characteristics. Human feeder–supported HES cells should provide a safer alternative to existing HES cell lines in therapeutic applications.

10/08/02: Scientists identify a new kind of genetic problem in muscular dystrophy

03/08/02: Stem cell discovery raises prospect of treating genetic disorders in the womb

03/08/02:Researchers find way to improve body's tolerance of stem cells

JULY

29/07/02: Expanded CUG Repeats Trigger Aberrant Splicing of ClC-1 Chloride Channel Pre-mRNA and Hyperexcitability of Skeletal Muscle in Myotonic Dystrophy

Ami Mankodi ⁵¹, Masanori P. Takahashi ⁵², Hong Jiang¹, Carol L. Beck³, William J. Bowers¹, Richard T. Moxley¹, Stephen C. Cannon², and Charles A. Thornton¹

In myotonic dystrophy (dystrophia myotonica, DM), expression of RNAs that contain expanded CUG or CCUG repeats is associated with degeneration and repetitive action potentials (myotonia) in skeletal muscle. Using skeletal muscle from a transgenic mouse model of DM, we show that expression of expanded CUG repeats reduces the transmembrane chloride conductance to levels well below those expected to cause myotonia. The expanded CUG repeats trigger aberrant splicing of pre-mRNA for ClC-1, the main chloride channel in muscle, resulting in loss of ClC-1 protein from the surface membrane. We also have identified a similar defect in ClC-1 splicing and expression in two types of human DM. We propose that a transdominant effect of mutant RNA on RNA processing leads to chloride channelopathy and membrane hyperexcitability in DM.

29/07/02: Researchers Identify Defect That Causes Rare Muscular Dystrophies

27/07/02: A Matter of Life and Breath - Dr. John Bach "Inspires" Patients to Survive and Thrive

27/07/02: Discovery Health Channel will air documentary focusing on Duchenne MD.

20/07/02: Dystrophin deficiency markedly increases enterovirus-induced cardiomyopathy: a genetic predisposition to viral heart disease

20/07/02: Successful bridge to transplantation in a patient with Becker muscular dystrophy-associated cardiomyopathy

20/07/02: Fukuyama-type congenital muscular dystrophy: a case report in the Japanese population living in Brazil

20/07/02: Facioscapulohumeral (FSHD1) and other forms of muscular dystrophy in the same family: is there more in muscular dystrophy than meets the eye?

12/07/02: MDA Scientists Find Two Chemicals That May Slow Wasting in Muscular Dystrophy

12/07/02: Blocking Myostatin Protein Could Treat Muscular Dystrophy

08/07/02: Parent Project: 2002 Annual Conference Abstracts

06/07/02: Peak Flow and Peak Cough Flow in the Evaluation of Expiratory Muscle Weakness and Bulbar Impairment in Patients with Neuromuscular Disease

Adrián Alejandro Suárez , MD ; Fernando Augusto Pessolano; Sergio Gabriel Monteiro, PT; Gabriela Ferreyra, PT; Maria Esther Capria, PT; Lilia Mesa, MD; Alberto Dubrovsky, MD; Eduardo Luis De Vito, MD

From the Instituto de Investigaciones Médicas Alfredo Lanari, Facultad de Medicina, Universidad de Buenos Aires, Argentina (AAS, FAP, SGM, GF, MEC, ELDV), and the Hospital Francés, Buenos Aires, Argentina (LM, AD).

AMERICAN JOURNAL OF PHYSICAL MEDICINE & REHABILITATION 2002;81:506-511

ABSTRACT

Suárez AA, Pessolano FA, Monteiro SG, Ferreyra G, Capria ME, Mesa L, Dubrovsky A, De Vito EL: Peak flow and peak cough flow in the evaluation of expiratory muscle weakness and bulbar impairment in patients with neuromuscular diseases. Am J Phys Med Rehabil 2002;81:506–511.

Objective: To study the expiratory muscle force and the ability to cough estimated by the peak expiratory flow and peak cough flow in patients with Duchenne muscular dystrophy and amyotrophic lateral sclerosis.

Design: A total of 27 patients with amyotrophic lateral sclerosis and 52 patients with Duchenne muscular dystrophy were studied. From the group of 144 normal subjects of this laboratory, we selected 38 for comparison.

Results: The maximal inspiratory pressure in patients with Duchenne muscular dystrophy and amyotrophic lateral sclerosis was 64.5 ± 24.7% and 37.8 ± 21.8%, respectively, and maximal expiratory pressure was 64.2 ± 32.5% and 37.7 ± 21.6%, respectively. Patient groups showed a significant lower peak expiratory flow than normal subjects. Higher peak cough flow than peak expiratory flow was found in all groups. The peak cough flow–peak expiratory flow difference was 46 ± 18% in normal subjects, 43 ± 23% in patients with Duchenne muscular dystrophy, and 11 ± 17% in patients with amyotrophic lateral sclerosis. The peak expiratory flow and peak cough flow were not different in bulbar onset amyotrophic lateral sclerosis. In patient groups, the dynamic and static behavior correlated positively.

Conclusions: These results suggest that peak cough flow–peak expiratory flow is useful to monitor expiratory muscle weakness and bulbar involvement and to assess its evolution in these patients.

06/07/02: Usefulness of Preoperative CK Levels as an MH Predictor in a High Risk Population

Bruce D. Levine, M.D. ; Elizabeth T. Drum, M.D.; Andrew Herlich, M.D.; Henry Rosenberg, M.D.

Department of Anesthesiology, Temple University School of Medicine, Philadelphia, Pennsylvania

Malignant Hyperthermia (MH) is difficult to predict preoperatively. High risk patients may include those with Duchenne's muscular dystrophy (DMD), scoliosis, osteogenesis imperfecta , and arthrogryposis. In the 1970s creatine kinase (CK) was used to identify patients at risk for MH. The specificity and sensitivity of CK was not sufficient to make this a useful screen. The Shriners Hospitals care for patients with orthopedic and musculoskeletal problems. At the Philadelphia Shriners, all patients undergoing surgical procedures requiring anesthesia have had preoperative CK levels measured since the 1970s. Initially, an elevated preop CK contraindicated a triggering anesthetic. By the 1980s, this guideline was not followed. The purpose of this study was to determine the incidence of MH in a population with a high prevalence of musculoskeletal disorders, and to determine the value of CK in predicting MH risk.Methods With IRB approval, 1453 records of surgical patients were reviewed. Demographics, diagnoses, medical history, CK levels, surgical procedures, anesthetics, and complications were noted. Additional review was carried out for: elevated preop CK, patients who received succinylcholine (sux), patients with an elevated CK level who received sux, and patients whose medical record indicated the possibility of MH.
Results:89.7% of patients had normal preop CK. 149 patients had an elevated CK (10.3%), 75 patients received sux (5.2%), 5 had high CK levels and received sux (0.34%), and 6 had the possibility of MH (0.41%).
Two patients had a confirmed MH or MH-like reaction. One had DMD, the other, Perthe's. Excluding the patient with the presumed diagnosis of MH in conjunction with DMD, the incidence of MH in this population is 1/1453= 0.069%. If the patient with known MH is included, the incidence is 2/1453=0.138%. Both are above the generally accepted rate of 1/50,000 (.002%)
Of 149 patients with an elevated CK, 140 received triggering agents. None experienced an MH or MH-like reaction. Of the nine who did not receive triggering agents, 4 had DMD, 1 arthrogryposis, 1 encephalopathy, 1 atlantoaxial subluxation, and 2 CP.
Of the 149 patients with an elevated CK, 28 (18.8%) had diagnoses of a musculoskeletal disorder: 18 scoliosis, 4 DMD, 2 Charcot-Marie-Tooth disease, 1 myotonic dystrophy, 1 spinocerebellar atrophy, and 1 arthrogryposis. Only the DMD patients and the arthrogrypotic received a non triggering anesthetic. Therefore of the 140 patients with elevated CK who received trigger agents(only one had sux), 23 had musculoskeletal disorders. None experienced an MH or MH-like reaction.
Conclusions:1. The incidence of MH in patients with musculoskeletal disorders is higher than the general population.2. Elevated CK in a population at higher risk for MH is not predictive of risk for MH or MH-like reactions. 3 The relationship between MH and musculoskeletal disorders other than DMD remains to be clarified.
Anesthesiology 2002; 96: A1237

02/07/02: Stable micro-dystrophin gene transfer using an integrating adeno-retroviral hybrid vector ameliorates the dystrophic pathology in mdx mouse muscle

02/07/02: Airway nitric oxide in Duchenne muscular dystrophy

JUNE

29/06/02: Right Ventricular MR Abnormalities in Myotonic Dystrophy and Relationship with Intracardiac Electrophysiologic Test Findings: Initial Results

29/06/02: Chimeric snRNA molecules carrying antisense sequences against the splice junctions of exon 51 of the dystrophin pre-mRNA induce exon skipping and restoration of a dystrophin synthesis in Delta 48-50 DMD cells

25/06/02: A Subpopulation of Murine Bone Marrow Cells Fully Differentiates along the Myogenic Pathway and Participates in Muscle Repair in the mdx Dystrophic Mouse

25/06/02: Overexpression of dystrobrevin delays locomotion defects and muscle degeneration in a dystrophin-deficient Caenorhabditis elegans

25/06/02: Altering Genes to Save Lives

22/06/02: Engraftment of bone marrow and fetal liver cells after in utero transplantation in MDX mice.

22/06/02: Effect of injecting primary myoblasts versus putative muscle-derived stem cells on mass and force generation in mdx mice

13/06/02: Pharmacological control of cellular calcium handling in dystrophic skeletal muscle

Urs T. Ruegg, Valerie Nicolas-Metral, Corinne Challet, Katy Bernard-Helary,

Olivier M. Dorchies, Stephanie Wagner, Timo M. Buetler

Abstract

Duchenne muscular dystrophy arises due to the lack of the cytoskeletal protein dystrophin. In Duchenne muscular dystrophy muscle, the lack of dystrophin is accompanied by alterations in the dystrophin–glycoprotein complex. We and others have found that the absence of dystrophin in cells of the Duchenne muscular dystrophy animal model, the mdx mouse, leads to elevated Ca21 influx and cytosolic Ca21 concentrations when exposed to stress. We have also shown that a-methylprednisolone, the only drug used successfully in the therapy of Duchenne muscular dystrophy, and creatine lowered cytosolic Ca21 levels in mdx myotubes. It is likely that chronic elevation of [Ca21] in the cytosol in response to stress is an initiating event for apoptosis and/or necrosis in Duchenne muscular dystrophy or mdx muscle and that alterations in mitochondrial function and metabolism are involved. Other cellular signalling pathways (e.g. nitric oxide) might also beaffected.

13/06/02: Integrin alpha 7 beta 1 in muscular dystrophy/myopathy of unknown etiology

11/06/02:Collaborative translational research leading to multicenter clinical trials in Duchenne muscular dystrophy: the Cooperative International Neuromuscular Research Group (CINRG)

Diana M. Escolar, Erik K. Henricson, Livia Pasquali, Ksenija Gorni, Eric Hoffman

Abstract

Progress in the development of rationally based therapies for Duchenne muscular dystrophy has been accelerated by encouraging multidisciplinary, multi-institutional collaboration between basic science and clinical investigators in the Cooperative International Research Group. We combined existing research efforts in pathophysiology by a gene expression profiling laboratory with the efforts of animal facilities capable of conducting high-throughput drug screening and toxicity testing to identify safe and effective drug compounds that target different parts of the pathophysiologic cascade in a genome-wide drug discovery approach. Simultaneously, we developed a clinical trial coordinating center and an international network of collaborating physicians and clinics where those drugs could be tested in large-scale clinical trials. We hope that by bringing together investigators at these facilities and providing the infrastructure to support their research, we can rapidly move new bench discoveries through animal model screening and into therapeutic testing in humans in a safe, timely and costeffective setting.

11/06/02: An effective, low-dosage, intermittent schedule of prednisolone in the long-term treatment of early cases of Duchenne dystrophy

Maria Kinali, Eugenio Mercuri, Marion Main, Francesco Muntoni, Victor Dubowitz

Abstract

The aim of this study was to evaluate the long-term effects in young children with Duchenne dystrophy of an intermittent low dosage regime of prednisolone (0.75 mg/kg per day for 10 days per month, or 10 days on and 10 days off). Six children under 5 years with Duchenne dystrophy have been commenced on this schedule, four of whom have been followed for at least 30 months and are reported here. All four presented with classical Duchenne dystrophy, and had an out-of-frame deletion in the Duchenne gene and absence of dystrophin in their muscle. All four showed a rapid and dramatic response in muscle function and strength. In three of the four there was an almost complete remission of all clinical signs of dystrophy. Their functional scores remained well above the average scores recorded in untreated Duchenne boys at the same age. There was no increase in weight, stunting of growth, decreased bone density or any other significant side effects related to the prednisolone. Our current experience suggests that this intermittent, low-dosage prednisolone regime is well tolerated and can be safely given long-term in young children with Duchenne dystrophy. The striking response also suggests that there may be an optimal window for treatment of Duchenne dystrophy in the early stages of the disease.

11/06/02: Glucocorticoid-mediated regulation of utrophin levels in human muscle fibers

Isabelle Courdier-Fruh, Lee Barman, Alexandre Briguet, Thomas Meier

Abstract

Previous studies on transgenic mice indicate that upregulation of utrophin protein may offer a potential treatment strategy for Duchenne muscular dystrophy. We have analyzed the effect of the glucocorticoid 6alfa-methylprednisolone-21 sodium succinate on utrophin protein levels using a cell-based assay with differentiated human myotubes derived from biopsies of healthy individuals or Duchenne muscular dystrophy patients. We found that within 5–7 days 6alfa-methylprednisolone-21 sodium succinate increases utrophin protein up to ,40% in both normal and dystrophin-deficient myotubes compared to untreated control cultures. When analyzed in promoter–reporter assays 6alfa-methylprednisolone- 21 sodium succinate activated a utrophin promoter A-fragment but did not activate a utrophin promoter B-fragment.Surprisingly, endogenous levels of utrophin mRNA in 6alfa-methylprednisolone-21 sodium succinate-treated muscle cells were unaltered indicating that the utrophin-inducing effect of glucocorticoids may be a result of post-transcriptional mechanisms. We have also analyzed 66 glucocorticoids for their effect on utrophin protein levels and found that glucocorticoids in general are able to induce utrophin protein in human myotubes.

08/06/02: Current protocol of a research phase I clinical trial of full-length dystrophin plasmid DNA in Duchenne/Becker muscular dystrophies Part II: clinical protocol

Norma Beatriz Romeroa, Olivier Benveniste, Christine Payan, Serge Braun, Patrick Squiban, Serge Herson, Michel Fardeau

Abstract

A phase I clinical study on gene therapy in Duchenne and Becker muscular dystrophy, open, without direct individual benefit for the patient, is being performed at the Pitie´-Salpeˆtrie`re Hospital, Paris. The aims of this project are: (a) to determine the tolerance and the safety of the intramuscular administration of dystrophin cDNA and (b) to study the quality of the gene transfer in vivo in human patients affected by Duchenne and Becker muscular dystrophy. This clinical trial is conducted sequentially and includes three cohorts of three patients each. Patients must be at least 15 years of age. Diagnosis of Duchenne and Becker muscular dystrophy was confirmed by molecular analysis of the dystrophin gene and for each patient the abnormal expression of dystrophin was confirmed, in skeletal muscle, with antibodies directed against the deleted part of the dystrophin. This phase I study is scheduled to be completed by the end of 2002.

08/06/02: Volunteers raise $40,880 for muscular dystrophy

08/06/02: Effect of zinc-carnosine chelate compound on muscle function in mdx mouse

06/06/02: Complement 3 deficiency and oral prednisolone improve strength and prolong survival of laminin alpha2-deficient mice

05/06/02: Abstracts from the Annual Meeting of American Society of Gene Therapy

01/06/02: Assessment of cardiac function in adolescents with Duchenne muscular dystrophy: importance of neurohormones

MAY

30/05/02: Adeno-associated virus vector gene transfer and sarcolemmal expression of a 144 kDa micro-dystrophin effectively restores the dystrophin-associated protein complex and inhibits myofibre degeneration in nude/mdx mice

26/05/02: Abstratcs from the European Society of Human Genetics

25/05/02: Corticosteroids in Duchenne muscular dystrophy: a reappraisal

23/05/02: Duchenne Muscular Dystrophy:Prolongation of Life by Noninvasive Ventilation and Mechanically Assisted Coughing

21/05/02: Identification of a novel population of muscle stem cells in mice : potential for muscle regeneration

18/05/02: Sp1 and the ets-related transcription factor complex GABP alpha/beta functionally cooperate to activate the utrophin promoter