HUMAN ARGINASE AND ARGINASE DEFICIENCY

本建议着重研究和开发两个克隆的 人类的辅酶A酶基因，首先是为了阐明其功能， 监管和进化，并最终作为一种手段， 替代患者中缺乏的腺苷酸酶（AI）活性的方法 伴有高维生素A血症和进行性神经和智力障碍 恶化 从小鼠中克隆的AI基因将用于创建AI缺陷型 敲除动物，其将被研究以确定 疾病生化和神经病理学，研究诱导 并最终作为基因或 自体酶替代疗法 将表征非肝转移酶基因（AIIs）的cDNA，并 用于分离编码它们的基因。这些试剂将 定义细胞生物学和AII以及基因的调控， 蛋白质产品和组织和功能的具体控制， 的位点 AI和AII将用作基因转移研究的试剂，以获得 更精确地界定这些监管控制的性质。这些 研究将利用异源控制下的AI和AII cDNA， 启动子，以及天然基因和调控基因的构建体， 序列和报告基因。通过分子操纵 信号序列，我们将探讨亚细胞的重要性， AI和AII生理功能的区室化， 基因治疗的新方法，通过改变转移基因的方向， 产品进出欧盟。这些研究将确定 使用AI、AII或AII基因的策略和组织特异性 两者的组合用于基因治疗，或用于AII的募集， 作为“自体酶替代”的手段。 这个项目利用了一个独特的模式，在一个理想的有利时机， 环境，研究代谢疾病的影响， 精神发育迟滞及其基因操作治疗。结果 这些研究应该证明与更多的疾病有关， 它的类型。

This proposal focuses on the study and exploitation of the two cloned arginase genes in man, first as a means of elucidating their function, regulation and evolution, and ultimately as a means of devising approaches to replacing the deficient arginase (AI) activity in patients with hyperargininemia and progressive neurological and intellectual deterioration. The cloned AI gene from mouse will be used to create an AI-deficient knockout animal which will be studied to define the pathogenesis of the disorder biochemically and neuropathologically, to study the induction of AII and ultimately as a test subject for molecular studies of gene or "autoenzyme replacement" therapy. cDNAs for non-liver arginase gene(s) (AIIs) will be characterized and be used to isolate the gene or genes encoding them. These reagents will define the cell biology and AII and the regulation of the gene and protein products and the tissue and function specific control of the loci. AI and AII will be used as reagents in gene transfer studies to obtain finer definition of the nature of these regulatory controls. These studies will utilize AI and AII cDNAs under the control of heterologous promotors, as well as constructs of the native gene and regulatory sequences, and reporter genes. Through the molecular manipulation of signal sequences, we will explore the importance of subcellular compartmentalization in the physiologic function of AI and AII and as novel approaches to gene therapy, by redirecting the transferred gene products into or out of the mitochondrion. These studies will define the strategy and tissue specificity for the use of the gene for AI, AII or a combination of the two for gene therapy, or for the recruitment of AII as a means of "autoenzyme replacement". This project exploits a unique model, at a propitious time in an ideal environment, to study the effects of a metabolic disease that causes mental retardation, and its treatment by gene manipulation. Results from these studies should prove relevant to a larger number of disorders of its type.