MUTANT ANALYSIS OF TSC1 & TSC2 IN TSC RELATED DISORDERS

本提案的目标是开发稳健、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，但也将成为分析TSC和相关疾病的分子病理学的有价值的研究工具。该项目开发的技术将直接应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。虽然它以常染色体显性模式遗传，但大多数新病例（约65%）是散发性的，没有先前的家族史。外显率高，但表达在严重和轻度受影响的个体中是可变的。最常见的器官是大脑、皮肤、肾脏和心脏。癫痫发作、智力迟钝和行为障碍引起的神经系统疾病很常见。最近发现了两种疾病基因，TSC1和TSC2。不幸的是，由于TSC中两个基因的大尺寸和突变谱的多样性，基因检测的发展受到了阻碍。该拨款的R21部分侧重于开发和优化一系列3种检测方法，这将允许对TSC进行高通量综合遗传分析。该提案的R33部分侧重于使用TSC的综合突变分析来收集大队列TSC患者的基因型和临床数据。综合突变检测方法也将用于研究TSC1和TSC2在相关疾病以及TSC病变和相关非TSC肿瘤中的作用。目前，TSC患者和家属对综合突变分析的需求很大，用于计划生育和产前诊断、诊断确认和预后信息。本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肿瘤分子病理中的作用。

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.