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.

描述（申请人提供）：表观可的松还原酶缺乏症（ACRD）是一种人类疾病，其中皮质醇的代谢清除增加，导致肾上腺皮质过度活动，肾上腺雄激素分泌增加，并具有类似于多囊卵巢的表型综合征（PCOS）/代谢综合征/X综合征。我们之前的工作表明 ACRD 是一种双基因疾病，需要携带两个基因突变， HSD11B1（编码 11β-羟基类固醇脱氢酶的 1 型或肝同工酶）和 H6PD（编码 6-磷酸己糖脱氢酶）。我们建议阐明这些突变相互作用导致疾病的机制。我们将通过使用定量 PCR 测量 mRNA 水平，并使用免疫印迹和酶活性测定测量蛋白质表达来定义 H6PD 在各种小鼠和人体组织中的表达。我们将通过在细菌中表达 H6PD 和 11β-HSD1，并通过分析不同的 H6PDH 表达对哺乳动物细胞中 11β-HSD1 氧化还原酶活性的影响，研究 H6PD 在确定 11β-HSD1 活性设定点中的直接作用。我们将确定 H6PD 活性变化对原代人类脂肪细胞和肝细胞表型的功能影响。我们将开发 ACRD 小鼠模型。为此，我们将产生一种 H6PD 定向失活的小鼠，并将其与具有失活 HSD11B1 等位基因的小鼠杂交。我们将培育双杂合突变小鼠以产生 ACRD 小鼠模型。表型表征将重点关注基因和蛋白质表达、肝脏和脂肪组织中的 11a-HSD1 活性和平衡设定点、肾上腺皮质的发育和调节类固醇生成的关键基因的表达水平、肾上腺功能测定、脂肪组织发育和肝酶表达。最后，我们将对患有多囊卵巢综合症的女性进行基因分型，以确定人类 HSD11B1 和 H6PDH 基因的多态性在多大程度上是导致这种情况的危险因素。基因分型数据的子集将作为关联研究、受影响的同胞对研究以及传递不平衡测试进行分析。将在 PCOS 受试者中寻找这些基因的其他多态性。