pathogenic mutation in genes associated with NM. Given the atypical presentation not attributable to a known NM gene, linkage analysis was performed on DNA samples from the four affected members along with whole genome sequencing (WGS) on mother’s DNA (proband). On bioinformatic analysis, she was identified to carry a heterozygous muta- tion c.1261C>T in the NEFL gene predicted to cause premature termi- nation of the transcript and a truncated neurofilament light polypeptide protein (p.R421X). This mutation was not present in dbSNP Build 137 or Exome variant server, Washington University, and it was confirmed to be present in all the four affected members of the family by Sanger sequencing. Further, Autosomal dominant mutations in NEFL are associated with Charcot–Marie–Tooth (CMT) disease, sub- types CMT2E/CMT1F. Most of the pathogenic NEFL mutations are missense changes postulated to cause dominant negative effects. Quanti- tative real-time PCR was performed on cDNA, extracted from muscle samples from an affected family member and an age-matched control, and the mutant transcript was found to be stable. We hypothesize that the mutant transcript does not undergo nonsense-mediated decay, thereby encoding a truncated protein that interferes with the function of the wild type NEFL protein. Further work is underway to understand the molecular basis of the disease. This finding further expands the spec- trum of NEFL mutation-associated phenotypes to NM. http://dx.doi:10.1016/j.nmd.2013.06.518 P.9.6 Combined cap disease and nemaline myopathy in the same patient caused by an autosomal dominant mutation in the TPM3 gene E. Malfatti 1 , U. Schaeffer 2 , F. Chapon 3 , Y. Yang 4 , B. Eymard 5 , R. Xu 4 , J. Laporte 2 , N.B. Romero 1 1 Unite ´ de Morphologie Neuromusculaire, Institut de Myologie, Groupe Hospitalier Universitaire La Pitie ´-Salpe ˆtrie `re, Paris, France; 2 Department of Translational Medicine and Neurogenetics, IGBMC, Illkirch, France; 3 CHU de Caen, Centre de Compe ´tences des Pathologies Neuro- musculaires, Co ˆ te de Nacre, Inserm U 1075 COMETE, Caen, France; 4 BGI-Shenzhen, Shenzhen, China; 5 Centre de re ´fe ´ rence de Pathologie Neuromusculaire Paris-Est, Institut de Myologie, GHU La Pitie ´-Salpe ˆtrie `re, Assistance Publiqu, Paris, France The slow alpha-tropomyosin gene (TPM3) has been associated with three distinct histological entities: nemaline myopathy (NM, NEM1), con- genital fibres type disproportion (CFTD), and cap disease (CD). Here we describe a 34-year-old French Caucasian patient who pre- sented global hypotonia from the age of three months. His examination at 19 years showed a long narrow face, bilateral ptosis, and high arched palate. He also presented diffuse axial weakness associated with some degree of upper limb girdle and distal lower limbs weakness. A right deltoid muscle biopsy effectuated at 19 years showed the pres- ence of well separated cap structures and clusters of nemaline bodies. Elec- tron microscopy (EM) confirmed the presence of characteristic cap structures and typical nemaline bodies. Exome sequencing analysis allowed us to identify a de novo missense mutation (p.Arg168Cys) in the TPM3 gene. With this report we describe a never reported association of combined cap disease and nemaline myopathy in a patient presenting a de novo mutation in the TPM3 gene. Our study enlarges the morphological spec- trum TPM3-related myopathies, and proves that cap and nemaline bodies are two sides of the same ‘coin’. http://dx.doi:10.1016/j.nmd.2013.06.519 P.9.7 Skeletal muscle biopsy reappraisal in nebulin-related nemaline myopathy E. Malfatti 1 , V.L. Lehtokari 2 , U. Schaeffer 3 , J. Bohm 3 , B. Estournet 4 , S. Quijano-Roy 4 , S. Monges 5 , F. Lubieniecki 5 , R. Bellance 6 , A.L. Taratuto 5 , B. Eymard 7 , M. Fardeau 1 , C. Wallgren-Pettersson 8 , J. Laporte 3 , N.B. Romero 1 1 Unite ´ de Morphologie Neuromusculaire, Institut de Myologie, Groupe Hospitalier Universitaire La Pitie ´-Salpe ˆtrie ` re, Paris, France; 2 Department of Medical Genetics, Haartman Institute, University of Helsinki, and the Folkha ¨ lsan Institute of Genetics, Biomedic, Helsinki, Finland; 3 Dept. Translational Medecine, IGBMC, Illkirch, Strasbourg, France; 4 AP-HP, Service de Pe ´diatrie, Ho ˆ pital Raymond Poincare’, Ho ˆ pitaux Universitaires Paris-Ile-de-France Ouest, Po ˆle pe ´ diat, Garches, France; 5 Hospital Nac- ional de Pediatrı ´a J.P Garrahan, and Instituto de Investigaciones Neuro- logicas, FLENI, Buenos Aires, Argentina; 6 CHU de Fort de France, Martinique, Fort-de-France, France; 7 Centre de re ´fe ´rence de Pathologie Neuromusculaire Paris-Est, Institut de Myologie, GHU La Pitie ´-Sal- pe ˆtrie `re, Assistance Publiqu, Paris, France; 8 Department of Medical Genetics, Haartman Institute, University of Helsinki, and the Folkha ¨ lsan Institute of Genetics, Biomedicum, Helsinki, Finland Nemaline myopathy (NM) is a congenital disorder associating muscle weakness with rods in muscle biopsy. Clinical presentation is variable spanning from severe to mild forms. Seven genes are associated with NM. The nebulin (NEB) gene is the most commonly mutated accounting for 50% of cases. We performed a muscle morphological analysis of thir- teen NEB-mutated patients with different clinical forms to define patho- logical patterns and clinical-morphological correlations. Four groups were identified according to clinical severity and age at biopsy. Group 1 (n = 4) comprises severe/lethal NM and biopsy in first days. Group 2 (n = 3) comprehends severe NM and biopsy after one month. Group 3 (n = 3) comprises typical NM and biopsy in childhood, and group 4 (n = 2) patients with mild NM and biopsy in adulthood. Molec- ular diagnosis was done using dHPLC/Sanger-sequencing in six patients and next-generation sequencing in seven. Biopsies underwent histoenzy- mological, immunohistochemical and ultrastructural analysis. Fiber type distribution, rod characteristics, distribution and localization were investi- gated. All patients presented NEB mutations consistent with AR inheri- tance. G1 showed type 2 predominance and scattered squared rods in 1/ 3 of fibers. Ultrastructural analysis revealed high percentage of fibers with sarcomeric disarray. G2 showed a variable pattern of fiber type distribu- tion spanning from slight type 2 predominance to type 1 uniformity. Rods presented variable distribution and shape. Ultrastructural analysis revealed rare fibers with sarcomeric disarray. In contrast, G3 and G4 pre- sented a homogeneous type 1 uniformity associated with well-delimited subsarcolemmal and/or cytoplasmic elongated rods without sarcomeric alterations. In conclusions we (1) identified an unreported association of type 2 predominance in muscle biopsy of patients presenting severe NEB-related NM; (2) suggest a direct correlation between the association of sarcomeric disarray and clinical severity. http://dx.doi:10.1016/j.nmd.2013.06.520 P.9.8 Nemaline myopathy: Mutations in alternatively spliced exons of the nebulin gene V.L. Lehtokari 1 , J. Laitila 2 , M. Hanif 1 , K. Pelin 3 , C. Wallgren- Pettersson 1 1 Folkha ¨ lsan Institute of Genetics and University of Helsinki, Faculty of Medicine, Medical Genetics, Helsinki, Finland; 2 Folkha ¨ lsan Institute of Genetics and University of Helsinki, Faculty of Biological and Environmen- tal Sciences, Biosciences, Helsinki, Finland; 3 University of Helsinki, Faculty of Biological and Environmental Sciences, Biosciences, Helsinki, Finland Abstracts / Neuromuscular Disorders 23 (2013) 738–852 785