Biochemical Basis of Cortisone Reductase Deficiency

可的松还原酶缺乏症的生化基础

• 批准号: 6941665
• 负责人: PERRIN C WHITE
• 金额: $ 29.81万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6941665
• 关键词: alcohol dehydrogenase; clinical research; cortisone; disease /disorder etiology; disease /disorder model; endocrine disorder; family genetics; female; gene expression; genetic polymorphism; genetic regulation; genetic susceptibility; genetically modified animals; hormone regulation /control mechanism; human genetic material tag; human tissue; hydroxysteroid dehydrogenases; laboratory mouse; model design /development; molecular pathology; patient oriented research; polycystic ovary syndrome; protein structure function; siblings

DESCRIPTION (provided by applicant): Apparent cortisone reductase deficiency (ACRD) is a human disease in which the metabolic clearance of cortisol is increased, leading to over activity of the adrenal cortex, increased secretion of adrenal androgens, and a phenotype similar to polycystic ovary syndrome (PCOS)/metabolic syndrome/syndrome X. Our previous work has shown that ACRD is a digenic disease, requiring carriage of mutations in two genes, HSD11B1 (encoding the type 1 or liver isozyme of 11beta-hydroxysteroid dehydrogenase) and H6PD (encoding hexose-6-phosphate dehydrogenase). We propose to elucidate the mechanisms by which these mutations interact to cause disease. We will define the expression of H6PD in a variety of murine and human tissues by measuring mRNA levels using quantitative PCR, and by measuring protein expression using immunoblotting and assays of enzymatic activity. We will investigate the direct role of H6PD in determining the set point of 11beta-HSD1 activity by expressing H6PD and 11beta-HSD1 in bacteria, and by analyzing the effect of varying H6PDH expression upon 11beta-HSD1 oxo-reductase activity in mammalian cells. We will determine functional consequences of variations in H6PD activity upon phenotype of primary human adipocytes and hepatocytes. We will develop a mouse model of ACRD. To do this, we will produce a mouse with a targeted inactivation of H6PD, and cross this to a mouse with an inactivated HSD11B1 allele. We will breed double heterozygous mutant mice to produce a mouse model of ACRD. Phenotypic characterization will focus on gene and protein expression, 11a-HSD1 activity and equilibrium set-point in liver and adipose tissue, development of the adrenal cortex and levels of expression for key genes regulating steroidogenesis, assays of adrenal function, adipose tissue development, and hepatic enzyme expression. Finally, we will genotype women with polycystic ovary syndrome to determine the degree to which polymorphisms in the human HSD11B1 and H6PDH genes are risk factors for the development of this condition. Subsets of the genotyping data will be analyzed as an association study, as an affected sib pair study, and by transmission disequilibrium testing. Additional polymorphisms in these genes will be sought in PCOS subjects.

DESCRIPTION (provided by applicant): Apparent cortisone reductase deficiency (ACRD) is a human disease in which the metabolic clearance of cortisol is increased, leading to over activity of the adrenal cortex, increased secretion of adrenal androgens, and a phenotype similar to polycystic ovary syndrome (PCOS)/metabolic syndrome/syndrome X. Our previous work has shown that ACRD is a digenic disease,需要在两个基因中运输突变，即HSD11B1（编码11Beta-羟基固醇脱氢酶的1型或肝同工酶）和H6PD（编码六糖-6-磷酸盐脱氢酶）。我们建议阐明这些突变相互作用以引起疾病的机制。我们将通过使用定量PCR测量mRNA水平以及使用免疫印迹和酶活性测定法测量蛋白质表达来定义H6PD在多种鼠和人体组织中的表达。我们将通过在细菌中表达H6PD和11BETA-HSD1来确定H6PD在确定11BETA-HSD1活性的设定点中的直接作用，并通过分析H6PDH表达对哺乳动物细胞中的H6PDH表达对11BETA-HSD1 OXO-还核酶活性的影响。我们将确定H6PD活性变化对原代人脂肪细胞和肝细胞表型的功能后果。我们将开发ACRD的鼠标模型。为此，我们将产生一种具有靶向灭活H6PD的小鼠，并将其跨到具有灭活的HSD11B1等位基因的小鼠。我们将繁殖双重杂合突变小鼠，以产生ACRD的小鼠模型。表型表征将集中在肝脏和脂肪组织中的基因和蛋白质表达，11A-HSD1活性以及平衡的设定点，肾上腺皮质的发展以及调节类固醇生成的关键基因的表达水平，肾上腺功能的测定，脂肪组织的发育，脂肪组织的发育和肝乙酰基表达。最后，我们将患有多囊卵巢综合征的基因型女性确定人类HSD11B1和H6PDH基因中的多态性是该疾病发展的风险因素。基因分型数据的子集将作为一项关联研究，受影响的SIB对研究以及通过传播不平衡测试分析。这些基因中的其他多态性将在PCOS受试者中寻求。