GENETIC DISEASES--A STRATEGY TO DEVELOP ANIMAL MODELS

遗传性疾病囊性纤维化(CF)，主要由 使肺部易受慢性感染，目前是一个主要的 体细胞基因治疗的候选人。然而，慢性萎缩性胃炎的小鼠模型 不会发展为肺部疾病，也不存在其他的CF动物模型。其他 动物模型在测试治疗方法方面可能有相当大的用处。 该提案将评估寻找此类模型的一般方法。 基于聚合酶链式反应单链的基因筛查 构象多态和异源双链分析(PCR-SSCP/HA)，将 用于检验两个假设：(1)DNA序列变异(突变+ Cftr基因内的多态)在非 人类灵长类动物和人类种群，以及(2)整个人类 群体，疾病导致CFTR突变的总频率 (减去DeltaF508之后)，提供预期的粗略估计 非人灵长类动物的突变频率。第二条的确认 假设将需要检测到一个或多个灵长类动物 Cftr基因中一个致病突变的杂合子。在人类身上， 不同人群中突变的综合载频率 除DeltaF508(迄今已发现约500个突变)外 大约相当于1/85到1/150的疾病频率 1/3万到1/90,000。拟议的研究计划旨在 在载波频率上检测到突变的可能性约为95% 相当于1/100万的发病率。当灵长类动物有 确定了一个染色体上的候选CF突变，我们评估了 突变对CFTRs转运和功能的影响。如果突变 扰乱CFTR功能，将启动育种计划以产生 CF纯合子灵长类的可再生种群。

The genetic disease cystic fibrosis (CF), which causes death primarily by predisposing the lungs to chronic infections, is presently a leading candidate for somatic gene therapy. However, the mouse model of CF does not develop lung disease, and no other animal model for CF exists. Other animal models could be of considerable usefulness for testing therapies. This proposal will evaluate a general approach for finding such models. Genetic screening, based on polymerase chain reaction single strand conformation polymorphism and heteroduplex analysis (PCR-SSCP/HA), will be used to test two hypotheses: (1) that DNA sequence variation (mutations + polymorphisms) within the CFTR gene is approximately equivalent in non- human primate and human populations, and (2) that across human populations, the aggregate frequency of disease causing mutations in CFTR (after subtraction of DeltaF508), provides a rough estimate of expected mutation frequencies in non-human primates. Confirmation of the second hypothesis will require the detection of one or more primates who are heterozygous for a disease-causing mutation in the CFTR gene. In humans, the combined carrier frequency rate in various populations for mutations other than DeltaF508 (about 500 mutations identified to date) is estimated to be about 1/85 to 1/150 equivalent to disease frequencies of about 1/30,000 to 1/90,000. The proposed research program is designed to have an about 95% chance of detecting a mutation at carrier frequencies corresponding to a disease rate of 1/1,000,000. When primates with candidate CF mutations on one chromosome are identified, we assess the effects of the mutation on CFTR trafficking and function. If the mutation disrupts CFTR function, a breeding program will be initiated to produce a renewable population of CF homozygous primates.