ANIMAL MODEL OF ARGINASE II DEFICIENCY

本提案的重点是对第二种辅酶Ⅱ酶基因（AII）的研究， 从两栖动物到人类的真核生物中发现， 了解其在精氨酸稳态中的作用，其调节和 最终利用它来替代有缺陷的肝脏 进行性高维生素血症患者中的辅酶Ⅱ酶（AI）活性 神经和智力退化 AII的cDNA和特异性抗体将用于研究AII 调节前列腺、肾脏、泌乳乳腺和巨噬细胞， 器官和细胞类型，其中AII表达特别高。 将作出特别努力，以确定具体的细胞类型， AII突出的生理环境，如巨噬细胞 激活或前列腺成熟与青春期。 的关系 一氧化氮合成酶和精氨酸脱羧酶 共享精氨酸作为底物，也将进行研究。 所有的老鼠 cDNA克隆将用于在129Sv中克隆和表征该基因 小鼠 这些知识将用于创建AII缺陷敲除 动物来定义它的表型，来定义基因的功能 产物，以了解其敲除对精氨酸的影响 体内平衡，并确定其对AI和其他 （如果存在的话）。 最终，各种缺陷AI和AII基因的组合发展起来 从AI和AII部分或全部缺陷动物的杂交中， 提供原材料，概念化一系列战略，从 基因激活到基因治疗，用于治疗AI（最终 所有的缺陷。

This proposal focuses on the study of the second arginase gene (AII), found in eucaryotes from amphibians to man, with the goal of understanding its role in arginine homeostasis, its regulation and ultimately in exploiting it as a means of replacing deficient liver arginase (AI) activity in patients with hyperargininemia and progressive neurological and intellectual deterioration. The cDNA and specific antibodies to AII will be used to study AII regulation in prostate kidney, lactating mammary gland and macrophages, organs and cell types in which AII expression is particularly high. Special efforts will be made to define specific cell types and physiological circumstances in which AII is prominent, such a macrophage activation or prostate maturation with puberty. The relationship between AII, nitric oxide synthetase and arginine decarboxylase, enzymes sharing arginine as a substrate, will be studied as well. The mouse AII cDNA clone will be used to clone and characterize the gene in 129Sv mice. This knowledge will be used to create an AII-deficient knockout animal to define its phenotype, to define the functions of the gene product, to understand the impact of its knockout on arginine homeostasis and to define its impact on the expression of AI and other arginases (if they exist). Ultimately various combinations of deficient AI & AII genes developed from crosses of AI and AII partially or wholly deficient animals will provide the raw material to conceptualize a series of strategies, from gene activation to gene therapy, for the treatment of AI (and ultimately AII) deficiencies.