MUTANT ANALYSIS OF TSC1 & TSC2 IN TSC RELATED DISORDERS

这项建议的目标是开发稳健、敏感和高通量的方法，用于肿瘤抑制综合征结节性硬化症(TSC)的突变分析。这些方法将立即用于临床目的，但也将是分析TSC和相关疾病的分子病理学的宝贵研究工具。在该项目期间开发的技术也将直接适用于其他疾病的遗传变异研究。TSC是一种以多器官良性肿瘤(错构瘤)发展为特征的家族性肿瘤综合征。虽然它是以常染色体显性遗传模式遗传的，但大多数新病例(约65%)是零星的，没有既往家族史。外显率高，但表达在严重和轻度影响的个体中都是不同的。最常受累的器官是大脑、皮肤、肾脏和心脏。癫痫、精神发育迟滞和行为障碍引起的神经系统疾病很常见。最近发现了两个致病基因TSC1和TSC2。不幸的是，这两个基因的大小和TSC突变谱的多样性阻碍了基因测试的发展。这笔赠款的R21部分侧重于开发和优化一系列3种分析方法，这些方法将允许对TSC进行高通量综合遗传分析。这项建议的R33部分侧重于对TSC进行全面的突变分析，以收集大量TSC患者的基因和临床数据。全面的突变检测方法也将用于研究TSC1和TSC2在相关疾病以及TSC病变和相关的非TSC肿瘤中的作用。目前，TSC患者和家庭对用于计划生育和产前诊断、诊断确认和预后信息的综合突变分析有很大的需求。拟议的研究计划将解决这一问题，并有助于阐明TSC1和TSC2在TSC病变和非TSC肿瘤的分子病理学中的作用。

The goals of this proposal are to develop robust, sensitive, and high throughput methods for mutation analysis for the tumor suppressor syndrome, tuberous sclerosis complex (TSC). These methods will be immediately useful for clinical purposes but will also be valuable research tools for analysing the molecular pathology in TSC and related disorders. The technology developed during this project will be directly applicable to the study of genetic variation in other disorders as well. TSC is a familial tumor syndrome characterized by the development of benign tumors (hamartomas) in multiple organs. Although it is inherited in an autosomal dominant pattern, the majority of new cases (about 65 percent) are sporadic without antecedent family history. Penetrance is high but expression is variable with both severely and mildly affected individuals. Organs most frequently involved are the brain, skin, kidneys and heart. Neurologic morbidity from seizures, mental retardation and behavior disorders is common. Two disease genes, TSC1 and TSC2 have been identified recently. Unfortunately the development of a genetic test has been hindered by the large size of the two genes and the diversity of the mutation spectrum in TSC. The R21 portion of this grant focuses on the development and optimization of a series of 3 assays which will allow for high throughput comprehensive genetic analysis for TSC. The R33 portion of this proposal focuses on using comprehensive mutation analysis for TSC to collect genotype and clinical data on a large cohort of TSC patients. Comprehensive mutation detection methods will also be used to study the role of TSC1 and TSC2 in related disorders as well as in TSC lesions and related non-TSC tumors. There is currently much demand from TSC patients and families for comprehensive mutation analysis for family planning and prenatal diagnosis, diagnosis confirmation, and prognostic information. The proposed plan of study will address this issue as well as help elucidate the role of TSC1 and TSC2 in the molecular pathology of TSC lesions and non-TSC tumors.