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NMP018 Local expression of donor-derived dystrophin after intramuscular injections of normal muscle-precursor cells in Duchenne muscular dystrophy patients
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Abstracts: Poster Presentations, the Seventh European Paediatric Neurology Society (EPNS) Congress
NMP018 Local expression of donor-derived dystrophin after intramuscular injections of normal muscle-precursor cells in Duchenne muscular dystrophy patients D. Skuk1 *, M. Goulet1 , B. Roy1 , J.-P. Bouchard2 , P. Chapdelaine1 , M. Sylvain3 , R. Roy4 , F.J. Dugre´ 4 , J.P. Tremblay1 . 1 Unit of Research in Human Genetics, Hospital Center of the University Laval, 2 Department of Neurology, Hospital of Child-Jesus, 3 Department of Pediatric Neurology, Hospital Center of the University Laval, 4 Research Center in Rheumatology and Immunology, Hospital Center of the University Laval, Quebec, Canada Purpose: Transplantation of muscle-precursor cells could be a therapeutic tool in the treatment of Duchenne muscular dystrophy (DMD). In a recent clinical trial, we tested in DMD patients those conditions that we used successfully in monkey experiments: (1) cell delivery by a protocol of high-density intramuscular injections, and (2) tacrolimus immunosuppression. Methods: Nine DMD patients received muscle-precursor cells (cultured from muscle biopsies of normal donors) by 25 or 100 parallel injections in 1 cm3 or less of the Tibialis anterior. Saline injections were performed similarly in the contralateral muscle. Tacrolimus was used for immunosuppression. Muscle biopsies were performed at the injected sites 4 weeks later. Results: We observed donor-dystrophin-positive myofibers or a significant increase of dystrophin-positive myofibers only in the cell-grafted sites of 8 patients. Donor-dystrophin-positive myofibers varied from 3 to 26% for the total number of myofibers in the biopsies. Donor-dystrophin transcripts or a significant increase of dystrophin transcripts were detected by RT-PCR only in the cell-grafted sites of the 9 patients. Conclusions: Significant donor-derived dystrophin expression can be obtained in the skeletal muscles of DMD patients following specific conditions of cell delivery (high-density intramuscular injections) and an appropriate control of acute rejection. NMP019 The use of the EK scale in the management of Duchenne muscular dystrophy C.P. White1 *, V. Lucas2 . 1 Morriston Hospital, Swansea, 2Wales National School of Medicine, UK Increased life expectancy and the high perceived quality of life of non-ambulant patients with DMD have increased the importance of longterm monitoring and management. The EK scale was developed in 2001 as a measure of functional ability in the non-ambulant stage of DMD. We aim to ascertain whether the EK scale can be used as a predictive tool for respiratory and cardiac complications. The case notes of 19 non ambulant boys with DMD were reviewed retrospectively, extracting information from clinic visits over the last two years. The EK sum is significantly related to time after loss of ambulation (P < 0.001) and FVC% (P < 0.001) but not to left ventricular function (P > 0.05). Patients with scoliosis had significantly higher EK sums (P < 0.002). Mean EK sums for patients 5 years after loss of ambulation with and without scoliosis were 13 and 7 respectively. All patients with an FVC < 20% predicted and an EK sum 15 were ventilated at night. The EK scale mirrors the functional decline of these boys once non-ambulant and their respiratory decline. It is unrelated to their cardiac status. It may be used as another indicator of the impending need for ventilation.
NMP020 Genetic data on muscular dystrophies in Hungarian patients V. Karcagi, H. Piko, J. Balog, B. Nagy, A. Herczegfalvi. 1 Dept. of Molecular Genetics and Diagnostics, National Institute of Environmental Health, 2 Dept. of Neurology, Bethesda Children’s Hospital, Budapest, Hungary, 3 Friedrich-Baur-Institute and Dept. of Neurology, LMU, Munich, Germany Muscular dystrophies are rare inherited disorders of skeletal muscle which form a clinically and genetically heterogeneous group. Since the discovery of the dystrophin protein involved in Duchenne/Becker muscular dystrophy, several other protein dysfunctions have been described in the dystrophin related glycoprotein complex of the sarcolemma. In the last few years, we introduced the genetic analysis of several such diseases in Hungary to provide exact diagnosis for the affected patients. For the analysis of Duchenne/Becker muscular dystrophy, to supplement the multiplex PCR detection of the most common dystrophin gene deletions, cDNA probes were used in order to detect carrier status and to delimit exact deletion borders. In addition, the new multiplex ligation-dependent probe amplification (MLPA) technique was introduced that enabled the examination of the entire dystrophin gene and the efficient screening of the carrier status. During the last years, 141 affected persons were screened and 77 had deletions whereas out of the 86 female relatives 39 proved to be carrier of the mutations. In another severe and relatively frequent disorder, the facioscapulohumeral muscular dystrophy D4Z4 repeat deletions are detected by Southern blot analysis using the p13E11 probe. For the confirmation of the clinical diagnosis 77 patients were genetically analyzed and 51 positive cases were detected, whereas out of the 43 asymptomatic family members 5 carried the FSHD mutation. The differential diagnosis of the heterogeneous LGMD group requires first the specific analysis of the dystrophinassociated glycoproteins. In total 56 immunohistochemical and 20 Western blot analyses have been performed in the German laboratories. In 26 patients the exact pathogenic mutation has been identified by PCR/RFLP (in Hungary) or DNA sequencing (in Germany). All these achievements help us to provide a better genetic service for the patients and to give proper genetic counseling, including the possibility of prenatal analyses, for the affected families. NMP021 Duchenne muscular dystrophy (DMD) genetics in the Red Cross hospital database K. Donald *, J. Wilmshurst. School of Adolescent and Child Health, Red Cross Children’s Hospital, Cape Town, South Africa DMD is the most common form of muscular dystrophy. The condition is X-linked recessive. The gene deletion is identified in approximately 65% of patients. Previous published data from South Africa stated that boys of African ancestry had a lower gene deletion frequency than those of European or mixed ancestry, suggesting that boys with DMD of African ancestry had unique mutations not detectable using laboratory methods current at that time. Aim: To establish whether the frequency of gene deletions in DMD boys of African ancestry on our database differed from international norms. Method: All the boys in our service with clinically suspected DMD are offered DNA testing. If negative, they are offered a muscle biopsy. Boys with confirmed DMD (positive genetics, muscle biopsy or an affected family member with confirmed diagnosis) were identified and their demographics defined. Results: Thirty-two boys had a confirmed diagnosis: Eighteen had gene deletions in the DMD region (56%) and the remainder had confirmatory findings on biopsy.
Abstracts: Poster Presentations, the Seventh European Paediatric Neurology Society (EPNS) Congress
NMP018 Local expression of donor-derived dystrophin after intramuscular injections of normal muscle-precursor cells in Duchenne muscular dystrophy patients D. Skuk1 *, M. Goulet1 , B. Roy1 , J.-P. Bouchard2 , P. Chapdelaine1 , M. Sylvain3 , R. Roy4 , F.J. Dugre´ 4 , J.P. Tremblay1 . 1 Unit of Research in Human Genetics, Hospital Center of the University Laval, 2 Department of Neurology, Hospital of Child-Jesus, 3 Department of Pediatric Neurology, Hospital Center of the University Laval, 4 Research Center in Rheumatology and Immunology, Hospital Center of the University Laval, Quebec, Canada Purpose: Transplantation of muscle-precursor cells could be a therapeutic tool in the treatment of Duchenne muscular dystrophy (DMD). In a recent clinical trial, we tested in DMD patients those conditions that we used successfully in monkey experiments: (1) cell delivery by a protocol of high-density intramuscular injections, and (2) tacrolimus immunosuppression. Methods: Nine DMD patients received muscle-precursor cells (cultured from muscle biopsies of normal donors) by 25 or 100 parallel injections in 1 cm3 or less of the Tibialis anterior. Saline injections were performed similarly in the contralateral muscle. Tacrolimus was used for immunosuppression. Muscle biopsies were performed at the injected sites 4 weeks later. Results: We observed donor-dystrophin-positive myofibers or a significant increase of dystrophin-positive myofibers only in the cell-grafted sites of 8 patients. Donor-dystrophin-positive myofibers varied from 3 to 26% for the total number of myofibers in the biopsies. Donor-dystrophin transcripts or a significant increase of dystrophin transcripts were detected by RT-PCR only in the cell-grafted sites of the 9 patients. Conclusions: Significant donor-derived dystrophin expression can be obtained in the skeletal muscles of DMD patients following specific conditions of cell delivery (high-density intramuscular injections) and an appropriate control of acute rejection. NMP019 The use of the EK scale in the management of Duchenne muscular dystrophy C.P. White1 *, V. Lucas2 . 1 Morriston Hospital, Swansea, 2Wales National School of Medicine, UK Increased life expectancy and the high perceived quality of life of non-ambulant patients with DMD have increased the importance of longterm monitoring and management. The EK scale was developed in 2001 as a measure of functional ability in the non-ambulant stage of DMD. We aim to ascertain whether the EK scale can be used as a predictive tool for respiratory and cardiac complications. The case notes of 19 non ambulant boys with DMD were reviewed retrospectively, extracting information from clinic visits over the last two years. The EK sum is significantly related to time after loss of ambulation (P < 0.001) and FVC% (P < 0.001) but not to left ventricular function (P > 0.05). Patients with scoliosis had significantly higher EK sums (P < 0.002). Mean EK sums for patients 5 years after loss of ambulation with and without scoliosis were 13 and 7 respectively. All patients with an FVC < 20% predicted and an EK sum 15 were ventilated at night. The EK scale mirrors the functional decline of these boys once non-ambulant and their respiratory decline. It is unrelated to their cardiac status. It may be used as another indicator of the impending need for ventilation.
NMP020 Genetic data on muscular dystrophies in Hungarian patients V. Karcagi, H. Piko, J. Balog, B. Nagy, A. Herczegfalvi. 1 Dept. of Molecular Genetics and Diagnostics, National Institute of Environmental Health, 2 Dept. of Neurology, Bethesda Children’s Hospital, Budapest, Hungary, 3 Friedrich-Baur-Institute and Dept. of Neurology, LMU, Munich, Germany Muscular dystrophies are rare inherited disorders of skeletal muscle which form a clinically and genetically heterogeneous group. Since the discovery of the dystrophin protein involved in Duchenne/Becker muscular dystrophy, several other protein dysfunctions have been described in the dystrophin related glycoprotein complex of the sarcolemma. In the last few years, we introduced the genetic analysis of several such diseases in Hungary to provide exact diagnosis for the affected patients. For the analysis of Duchenne/Becker muscular dystrophy, to supplement the multiplex PCR detection of the most common dystrophin gene deletions, cDNA probes were used in order to detect carrier status and to delimit exact deletion borders. In addition, the new multiplex ligation-dependent probe amplification (MLPA) technique was introduced that enabled the examination of the entire dystrophin gene and the efficient screening of the carrier status. During the last years, 141 affected persons were screened and 77 had deletions whereas out of the 86 female relatives 39 proved to be carrier of the mutations. In another severe and relatively frequent disorder, the facioscapulohumeral muscular dystrophy D4Z4 repeat deletions are detected by Southern blot analysis using the p13E11 probe. For the confirmation of the clinical diagnosis 77 patients were genetically analyzed and 51 positive cases were detected, whereas out of the 43 asymptomatic family members 5 carried the FSHD mutation. The differential diagnosis of the heterogeneous LGMD group requires first the specific analysis of the dystrophinassociated glycoproteins. In total 56 immunohistochemical and 20 Western blot analyses have been performed in the German laboratories. In 26 patients the exact pathogenic mutation has been identified by PCR/RFLP (in Hungary) or DNA sequencing (in Germany). All these achievements help us to provide a better genetic service for the patients and to give proper genetic counseling, including the possibility of prenatal analyses, for the affected families. NMP021 Duchenne muscular dystrophy (DMD) genetics in the Red Cross hospital database K. Donald *, J. Wilmshurst. School of Adolescent and Child Health, Red Cross Children’s Hospital, Cape Town, South Africa DMD is the most common form of muscular dystrophy. The condition is X-linked recessive. The gene deletion is identified in approximately 65% of patients. Previous published data from South Africa stated that boys of African ancestry had a lower gene deletion frequency than those of European or mixed ancestry, suggesting that boys with DMD of African ancestry had unique mutations not detectable using laboratory methods current at that time. Aim: To establish whether the frequency of gene deletions in DMD boys of African ancestry on our database differed from international norms. Method: All the boys in our service with clinically suspected DMD are offered DNA testing. If negative, they are offered a muscle biopsy. Boys with confirmed DMD (positive genetics, muscle biopsy or an affected family member with confirmed diagnosis) were identified and their demographics defined. Results: Thirty-two boys had a confirmed diagnosis: Eighteen had gene deletions in the DMD region (56%) and the remainder had confirmatory findings on biopsy.