Department of Translational Medicine and Neurogenetics, Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, France.
PLoS One. 2013 Jun 24;8(6):e67527. doi: 10.1371/journal.pone.0067527. Print 2013.
Congenital myopathies are severe muscle disorders affecting adults as well as children in all populations. The diagnosis of congenital myopathies is constrained by strong clinical and genetic heterogeneity. Moreover, the majority of patients present with unspecific histological features, precluding purposive molecular diagnosis and demonstrating the need for an alternative and more efficient diagnostic approach. We used exome sequencing complemented by histological and ultrastructural analysis of muscle biopsies to identify the causative mutations in eight patients with clinically different skeletal muscle pathologies, ranging from a fatal neonatal myopathy to a mild and slowly progressive myopathy with adult onset. We identified RYR1 (ryanodine receptor) mutations in six patients and NEB (nebulin) mutations in two patients. We found novel missense and nonsense mutations, unraveled small insertions/deletions and confirmed their impact on splicing and mRNA/protein stability. Histological and ultrastructural findings of the muscle biopsies of the patients validated the exome sequencing results. We provide the evidence that an integrated strategy combining exome sequencing with clinical and histopathological investigations overcomes the limitations of the individual approaches to allow a fast and efficient diagnosis, accelerating the patient's access to a better healthcare and disease management. This is of particular interest for the diagnosis of congenital myopathies, which involve very large genes like RYR1 and NEB as well as genetic and phenotypic heterogeneity.
先天性肌病是一种严重的肌肉疾病,影响所有人群的成人和儿童。先天性肌病的诊断受到强烈的临床和遗传异质性的限制。此外,大多数患者表现出非特异性的组织学特征,排除了有针对性的分子诊断,这表明需要一种替代的、更有效的诊断方法。我们使用外显子组测序,辅以肌肉活检的组织学和超微结构分析,在 8 名具有不同临床骨骼肌病理的患者中鉴定出致病突变,从致命的新生儿肌病到轻度和缓慢进展的成人起病肌病。我们在 6 名患者中发现了 RYR1(ryanodine receptor)突变,在 2 名患者中发现了 NEB(nebulin)突变。我们发现了新的错义和无义突变,揭示了小的插入/缺失,并证实了它们对剪接和 mRNA/蛋白稳定性的影响。患者的肌肉活检的组织学和超微结构发现验证了外显子组测序的结果。我们提供的证据表明,将外显子组测序与临床和组织病理学研究相结合的综合策略克服了单一方法的局限性,能够快速有效地进行诊断,加速患者获得更好的医疗保健和疾病管理。这对于诊断涉及 RYR1 和 NEB 等大型基因以及遗传和表型异质性的先天性肌病尤其重要。