ANIMAL MODELS OF MUCOPOLYSACCHARIDOSES

粘多糖动物模型

基本信息

批准号：
3151547
负责人：
MARK E HASKINS
金额：
$ 23.79万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
1979
资助国家：
美国
起止时间：
1979-07-01 至 1988-06-30
项目状态：
已结题

项目摘要

The overall purpose of the proposed research is to use selected animal models of the human mucopolysaccharidoses to develop and evaluate various therapeutic strategies including enzyme, cellular and genetic manipulations. Inherent in these studies is the characterization of the molecular pathology of each animal model to determine the degree of homology to the respective human disease at the pathologic, biochemical and genetic levels. Four disorders, feline MPS I and VI; canine MPS III and VII, are under study and each has been or will be investigated according to the following scheme: Models are identified, breeding colonies established and the natural history and pathologic manifestations characterized. The enzyme defect is then characterized by purifying normal and residual enzyme and measuring the physiokinetic and immunologic properties of each. Accumulated substrate in various tissues is quantified and correlated with the morphologic pathology. The genetic defect is then characterized by sequencing the normal enzyme, synthesizing appropriate probes, screening cDNA and genomic libraries and comparing the structure and organization of the normal gene to that of the mutant gene. Three therapeutic strategies are evaluated: 1) Enzyme manipulation using compounds which enhance residual enzyme activity, 2) Cellular manipulation using allotransplantation of bone marrow, and 3) Somatic genetic manipulation using mammalian expression vectors to transfer full length cDNA clones into dificient cultured fibroblasts and, with appropriate viral vectors, perform in vivo transfection experiments. The effects of treatment on the clinical and pathologic course of the diseases, enzyme activity and substrate accumulation are evaluated to determine if therepy can prevent, arrest or reverse the disease process.

拟议研究的总体目的是使用选定的动物人类粘多糖糖的模型，以开发和评估各种治疗策略，包括酶，细胞和遗传操纵。这些研究固有的是表征每个动物模型的分子病理学以确定在病理学，生化和遗传水平。四个疾病，猫MP I和VI；犬MPS III和 vii，正在研究，每个人都已经或将根据以下方案：确定模型，建立繁殖菌落和自然历史和病理表现形式的特征。酶缺陷是然后以净化正常和残留酶并进行测量为特征每个人的生理动力学和免疫学特性。积累量化各种组织中的底物并与形态病理学。然后，遗传缺陷的特征是测序正常酶，合成适当的探针，筛选cDNA和基因组文库以及比较正常基因的结构和组织突变基因。评估了三种治疗策略：1）使用酶操作增强残留酶活性的化合物，2）细胞操作使用骨髓的同种异体移植，3）体细胞遗传使用哺乳动物表达载体操作以传递全长 cDNA克隆成可分培养的成纤维细胞，并具有适当的病毒向量，执行体内转染实验。效果疾病的临床和病理病程治疗酶评估活性和底物积累以确定是否可以预防，逮捕或扭转疾病过程。