Glyoxylate Reductase and Primary Hyperoxaluria

乙醛酸还原酶和原发性高草酸尿症

• 批准号: 7316284
• 负责人: ROSS P HOLMES
• 金额: $ 29.42万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7316284
• 关键词: Anions; Biological Assay; Blood; Calcium Oxalate; Calculi; Cell Line; Cells; Chromatography; Class; Collagen; Coupled; Cultured Cells; Data; Degradation Pathway; Deposition; Depth; Detection; Disease; Disruption; Enzymes; Erythrocytes; Excretory function; Functional disorder; Genes; Genomics; Glycolates; Glyoxylates; Goals; Hereditary Disease; Human; Hydroxyproline; Hydroxypyruvate reductase; Hyperoxaluria; Incubated; Individual; Inherited; Investigation; Ions; Kidney; Kidney Calculi; Kidney Failure; Kidney Transplantation; Knock-out; Knockout Mice; Knowledge; Lead; Left; Liquid substance; Liver; Measures; Medical; Metabolic; Metabolism; Modeling; Monitor; Mus; Mutation; Oxalates; Pathway interactions; Pharmaceutical Preparations; Phenotype; Prevention therapy; Primary Hyperoxaluria; Production; RNA Interference; Reporting; Research Personnel; Role; Source; Study models; Testing; Therapeutic; Tissues; Urination; Urine; Western Blotting; day; design; enzyme deficiency; glycolate; glyoxylate; glyoxylate reductase; hydroxyproline oxidase; insight; liver transplantation; novel; novel strategies; novel therapeutics; phosphoglycolate; programs; research study; tool; urinary

DESCRIPTION (provided by applicant): Primary hyperoxaluria is a rare, genetic disorder that results from an excessive endogenous synthesis of oxalate. Glyoxylate reductase (GR) is a key enzyme in this biosynthetic pathway, converting glyoxylate, the main precursor of oxalate, to glycolate and thereby limiting its conversion to oxalate. The long term goals of this project are to develop an in depth understanding of the pathways associated with GR activity and glyoxylate and oxalate synthesis, to determine the metabolic consequences of an absence of GR activity, and to identify therapeutic strategies that limit oxalate synthesis when the activity is absent. To assist with these goals we have developed a cell line with reduced GR activity and have obtained a knockout (KO) species of mouse lacking GR activity. This proposal has three specific aims. The first aim is to determine the phenotype of the KO mouse, measuring anion concentrations in blood, urine and tissues using ion chromatography coupled to mass detection (IC/MS). The second aim is designed to test the hypothesis that inhibiting hydroxyproline oxidase (HPOX) will be an effective therapeutic strategy for decreasing the increased oxalate synthesis associated with GR deficiency. To establish proof of principle, RNA interference will be utilized to inhibit HPOX activity and decrease oxalate synthesis. The KO mouse will be used as a model for these studies. In the third specific aim, the functional role of GR in red blood cells will be examined. These cells have a simplified metabolism in comparison to other cells and preliminary data indicate that these cells contain glycolate, glyoxylate and oxalate, as well as GR activity. It is hypothesized that GR functions to limit oxalate production and to synthesize glycolate, a potential substrate for phosphoglycolate synthesis. These hypotheses will be tested in cells from KO mice and human cells incubated with glycolate and glyoxylate. These studies will increase our understanding of the metabolism associated with oxalate synthesis and will potentially identify novel therapeutic strategies to decrease oxalate synthesis in primary hyperoxaluria.

描述（由申请人提供）：原发性高草酸尿症是一种罕见的遗传性疾病，由草酸盐过度内源性合成引起。乙醛酸还原酶 (GR) 是该生物合成途径中的关键酶，可将草酸的主要前体乙醛酸转化为乙醇酸，从而限制其转化为草酸。该项目的长期目标是深入了解与 GR 活性以及乙醛酸和草酸合成相关的途径，确定缺乏 GR 活性的代谢后果，并确定限制草酸合成的治疗策略。没有活动。为了帮助实现这些目标，我们开发了一种 GR 活性降低的细胞系，并获得了缺乏 GR 活性的敲除 (KO) 小鼠物种。该提案具有三个具体目标。第一个目标是确定 KO 小鼠的表型，使用离子色谱结合质谱检测 (IC/MS) 测量血液、尿液和组织中的阴离子浓度。第二个目标旨在检验以下假设：抑制羟脯氨酸氧化酶 (HPOX) 将是减少与 GR 缺乏相关的草酸盐合成增加的有效治疗策略。为了建立原理证明，将利用 RNA 干扰来抑制 HPOX 活性并减少草酸合成。 KO 小鼠将用作这些研究的模型。在第三个具体目标中，将检查 GR 在红细胞中的功能作用。与其他细胞相比，这些细胞具有简化的代谢，初步数据表明这些细胞含有乙醇酸、​​乙醛酸和草酸，以及 GR 活性。据推测，GR 的功能是限制草酸的产生并合成乙醇酸，乙醇酸是磷酸乙醇酸合成的潜在底物。这些假设将在来自 KO 小鼠的细胞和与乙醇酸和乙醛酸一起孵育的人类细胞中进行测试。这些研究将增加我们对与草酸盐合成相关的代谢的理解，并有可能确定减少原发性高草酸尿症草酸盐合成的新治疗策略。