肌营养不良基因诊断及新突变致病性研究

肌营养不良（MD）是一组遗传性肌肉疾病，综合分析症状、体征、生化、免疫组化、相关基因检测等结果是MD确诊、预后及遗传咨询的依据，本课题组前期运用上述方法对160个疑诊MD病人进行分析，在基因水平上证实149个先证者分别为8种不同的MD，1例确诊为非MD，但仍有10例病人的致病基因未知； 16个新突变的致病性通过家系共分离分析、生物信息学软件预测得到了支持，应用免疫组化、肌肉细胞体外培养、荧光定量PCR、T-A克隆、基因测序等方法进一步确认其中2个新突变的致病性，并初步探索了其致病机制；本课题组拟继续扩大研究对象，并引入基因置换、细胞转染、基因表达分析等遗传工程技术加深对新突变生物学功能及致病机理的研究；并运用全基因组测序、RNA测序等技术探索疑难病例的致病基因；总结广西地区不同类型MD病人基因突变特点，明确新突变的致病性，为精准遗传咨询乃至基因治疗提供理论依据。

Muscular dystrophy (MD) is a group of inherited muscular diseases. The clinical symptoms, signs, biochemistry, immunohistochemistry technology, especially the genetic detections are helpful to make the diagnosis, genetic counseling and prognosis for MD. We have collected 160 suspected patients. Genetic diagnoses have been made for 149 patients, 8 different types MD were found, and novo mutations were found in 16 patients. However, the genetic pathogenesis of 10 suspected patients are still unknown. Meanwhile, the pathogenicity for most of 16 novo mutations was analyzed and supported by cosegregation analysis, bioinformatics software. Moreover the pathogenicity and pathogenic mechanism of two novo mutations had been confirmed by the results of immunohistochemistry, cell culture, real time PCR, TA cloning and gene sequence. We plan to continue our study in a larger population and improve the diagnostic strategies. Gene replacement， cell transfection and genetic expression analysis will be used to analyze the pathogenesis of the novo mutations. As occasion requires, whole genome sequencing and RNA sequencing techniques will be used to search for possible pathogenetic gene. We aim to identify mutation hot spots of different forms of MD, and to learn the pathogenicity of novo mutations, so as to provide a theoretical basis for accurate genetic counseling and even gene therapy.