Goldyne Savad Institute of Gene Therapy, Hadassah Hebrew University Medical Center, Jerusalem, Israel.
PLoS One. 2013 Jul 24;8(7):e69296. doi: 10.1371/journal.pone.0069296. Print 2013.
We describe an autosomal recessive heterogeneous congenital myopathy in a large consanguineous family. The disease is characterized by variable severity, progressive course in 3 of 4 patients, myopathic face without ophthalmoplegia and proximal muscle weakness. Absence of cores was noted in all patients. Genome wide linkage analysis revealed a single locus on chromosome 19q13 with Zmax = 3.86 at θ = 0.0 and homozygosity of the polymorphic markers at this locus in patients. Direct sequencing of the main candidate gene within the candidate region, RYR1, was performed. A novel homozygous A to G nucleotide substitution (p.Y3016C) within exon 60 of the RYR1 gene was found in patients. ARMS PCR was used to screen for the mutation in all available family members and in an additional 150 healthy individuals. This procedure confirmed sequence analysis and did not reveal the A to G mutation (p.Y3016C) in 300 chromosomes from healthy individuals. Functional analysis on EBV immortalized cell lines showed no effect of the mutation on RyR1 pharmacological activation or the content of intracellular Ca(2+) stores. Western blot analysis demonstrated a significant reduction of the RyR1 protein in the patient's muscle concomitant with a reduction of the DHPRα1.1 protein. This novel mutation resulting in RyR1 protein decrease causes heterogeneous clinical presentation, including slow progression course and absence of centrally localized cores on muscle biopsy. We suggest that RYR1 related myopathy should be considered in a wide variety of clinical and pathological presentation in childhood myopathies.
我们描述了一个在一个大型近亲家庭中常染色体隐性异质性先天性肌病。该疾病的特征为病情严重程度不一,4 名患者中有 3 名呈进行性发展,存在无眼肌麻痹的肌病面容和近端肌无力,但无中央核。所有患者均未见中央核。全基因组连锁分析显示 19q13 染色体上存在单一基因座,θ = 0.0 时 Zmax = 3.86,患者在此基因座纯合多态性标记。对候选区域内的主要候选基因 RYR1 进行直接测序。在 RYR1 基因的第 60 外显子中发现了一个新的纯合 A 到 G 核苷酸取代(p.Y3016C)。使用 ARMS PCR 对所有可利用的家族成员和另外 150 名健康个体进行了突变筛查。该程序证实了序列分析,并且在 300 条来自健康个体的染色体中未发现 A 到 G 突变(p.Y3016C)。对 EBV 永生化细胞系的功能分析表明,该突变对 RyR1 药理学激活或细胞内 Ca(2+)储存含量没有影响。Western blot 分析表明,患者肌肉中的 RyR1 蛋白显著减少,同时 DHPRα1.1 蛋白减少。该新突变导致 RyR1 蛋白减少,引起异质性临床表现,包括进展缓慢且肌肉活检中不存在中央核。我们建议在儿童肌病中,应考虑将 RYR1 相关肌病作为各种临床表现和病理表现的原因。