Department of Pediatrics, Kobe University Graduate School of Medicine, Kobe, Japan.
J Hum Genet. 2010 Jun;55(6):379-88. doi: 10.1038/jhg.2010.49. Epub 2010 May 20.
Recent developments in molecular therapies for Duchenne muscular dystrophy (DMD) demand accurate genetic diagnosis, because therapies are mutation specific. The KUCG (Kobe University Clinical Genetics) database for DMD and Becker muscular dystrophy is a hospital-based database comprising 442 cases. Using a combination of complementary DNA (cDNA) and chromosome analysis in addition to conventional genomic DNA-based method, mutation detection was successfully accomplished in all cases, and the largest mutation database of Japanese dystrophinopathy was established. Among 442 cases, deletions and duplications encompassing one or more exons were identified in 270 (61%) and 38 (9%) cases, respectively. Nucleotide changes leading to nonsense mutations or disrupting a splice site were identified in 69 (16%) or 24 (5%) cases, respectively. Small deletion/insertion mutations were identified in 34 (8%) cases. Remarkably, two retrotransposon insertion events were also identified. Dystrophin cDNA analysis successfully revealed novel transcripts with a pseudoexon created by a single-nucleotide change deep within an intron in four cases. X-chromosome abnormalities were identified in two cases. The reading frame rule was upheld for 93% of deletion and 66% of duplication mutation cases. For the application of molecular therapies, induction of exon skipping was deemed the first priority for dystrophinopathy treatment. At one Japanese referral center, the hospital-based mutation database of the dystrophin gene was for the first time established with the highest levels of quality and patient's number.
近年来，杜氏肌营养不良症（DMD）的分子疗法取得了新的进展，这就要求进行准确的基因诊断，因为这些疗法具有突变特异性。KUCG（神户大学临床遗传学）数据库包含了 442 个 DMD 和贝克肌营养不良症的病例，是一个基于医院的数据库。除了传统的基于基因组 DNA 的方法外，我们还使用 cDNA 和染色体分析相结合的方法，成功地在所有病例中检测到了突变，并建立了日本最大的肌营养不良症相关抗肌萎缩蛋白基因突变数据库。在这 442 个病例中，分别有 270 例（61%）和 38 例（9%）存在一个或多个外显子的缺失和重复，69 例（16%）和 24 例（5%）存在导致无义突变或剪接位点破坏的核苷酸变化，34 例（8%）存在小的缺失/插入突变。值得注意的是，还发现了两个反转录转座子插入事件。通过对肌营养不良蛋白 cDNA 的分析，我们在 4 个病例中成功地揭示了由内含子中单核苷酸变化引起的假外显子产生的新型转录本。在 2 个病例中发现了 X 染色体异常。对于缺失和重复突变，阅读框规则分别得到了 93%和 66%的维持。为了应用分子疗法，我们认为外显子跳跃诱导是肌营养不良症治疗的首要任务。在日本的一个转诊中心，我们首次建立了基于医院的肌营养不良基因突变数据库，该数据库具有最高的质量和患者数量。
