H+-ATPase B-subunit Dysfunction and Calcium Nephrolithiasis

H-ATP酶 B 亚基功能障碍和钙肾结石

• 批准号: 7930519
• 负责人: Orson W Moe
• 金额: $ 39.25万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-15 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7930519
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; Address; Aldehyde-Lyases; Alleles; Anions; Binding; Biochemical; Biochemistry; Biology; Calcium; Calcium Oxalate; Calculi; Candidate Disease Gene; Carbonic Anhydrase II; Carrier State; Cataloging; Catalogs; Cells; Cellular biology; Chemicals; Chemistry; Clinical; Clinical Data; Code; Complex; Coupling; Data; Databases; Defect; Diagnosis; Disease; Distal renal tubular acidosis Type 1; Face; Family member; Foundations; Frequencies; Functional disorder; Gene Frequency; General Population; Genes; Genetic; Genetic Variation; Genotype; Gray unit of radiation dose; Heterozygote; Human; Hydrolysis; In Vitro; Individual; Inheritance Patterns; Inherited; Inpatients; Kidney; Kidney Calculi; Kidney Diseases; Laboratory Study; Low Prevalence; Mammalian Cell; Metabolic; Molecular; Mutation; Nephrolithiasis; Oxalates; Patients; Persons; Phenotype; Physiology; Plasma; Point Mutation; Polygenic Traits; Population; Predisposition; Process; Protein Biochemistry; Proteins; Proton Pump; Proton-Translocating ATPases; Pump; Risk; Risk Factors; Structure; System; Testing; Urine; Variant; Work; Yeasts; base; calcium phosphate; cohort; genetic variant; high risk; human data; hypercalciuria; inhibitor/antagonist; medical attention; mutant; population based; public health relevance; reconstitution; research study; vacuolar H+-ATPase

DESCRIPTION (provided by applicant): The hereditary component of nephrolithiasis is a complex polygenic trait which is a formidable challenge to unravel. The study of monogenic causes of nephrolithiasis has been most informative in uncovering genetic components of nephrolithiasis. Calcareous stones are common in patients with congenital distal renal tubular acidosis :dRTA; but sd RcoTnAs idered a very rare cause of calcium stones due to the low prevalence of the disease. Congenital dRTA is a disease of renal acidification caused by mutations in either the vacuolar :VPa;se-A T :B1 or a4 subunit;, anion exchanger-1, or carbonic anhydrase II. This proposal focuses on the B1 subunit of the V-ATPase. The heterozygous carrier state is universaly believed to be silent. The mechanisms by which a mutant B1 subunit cause pump dysfunction is not well understood. We will test 3 hypotheses in this proposal: I. Heterozygous mutations of B1 are not rare in the stone forming population. II. The burden of heterozygosity is not silent but confers propensity for or affliction by calcium phosphate or oxalate nephrolithiasis. III. Mutant B1 subunits cause pump dysfunction by multiple mechanisms such as defective pump assembly, impaired ATP binding/hydrolysis, and deranged coupling to HE translocation. We will test these hypotheses with 3 Aims: 1. Population-based studies: Define the allele frequency of all base changes in B1 in the stone forming and non-stone forming population and collect the corresponding clinical data of these individuals. 2. Inpatient human studies: Perform detailed metabolic and physicochemical studies of selected individuals heterozygous for B1 to characterize their phenotype. 3. Bench studies: Define the mechanisms of how various mutant B1 subunits cause HE pump dysfunction using cell biology and biochemistry. This is not a study of classical dRTA. Instead we may be able to prove that monogenic predisposition is actually not rare in the general stone-forming population and many stone formers may harbor B1 mutations that predispose them to calcium stones. This may set precedence, for other monogenic predispositions in the general stone forming population. These studies will also significantly advance the understanding of V-ATPase biology. These natural point mutations and their cognate mode of dysfunction will provide valuable structure-function information on the biology of the B1 protein. PUBLIC HEALTH RELEVANCE: Each human has a 10-20% chance of getting a kidney stone in his or her lifetime. If a person has a family member with kidney stones, that individual has a higher risk of getting kidney stones. This process can involve many genes but there are circumstances where single genes can put a person at risk for kidney stones such as in a condition is distal renal tubular acidosis (abbreviated as dRTA). Most patients with dRTA get calcium kidney stones, but most patients with calcium kidney stones do not have dRTA. This proposal examines some new ideas. Based on our work in the last year, we believe that a milder form of dRTA where one inherits one instead of two defective genes, termed "heterozygous state" may be quite common in the general population of calcium kidney stones formers. The gene in question codes for a protein called "B1" which in its abnormal state will change the composition in the urine resulting in kidney stone formation. We will use human population and laboratory studies to answer three questions. 1. Exactly how common are abnormal B1 in a heterozygous state in people with calcium kidney stones? 2. If one carries an abnormal copy of B1 (the heterozygous state), what exactly are the abnormalities in the urine that predispose that person to having kidney stones? 3. How does an abnormal B1 change the composition of our urine so kidney stones will form? These questions, when answered, help us understand the hereditary component of kidneys stones and how abnormalities in single proteins, such as B1, will change our urine composition in an unfavorable way allowing allow us to better predict, diagnose, and treat kidney stones.

描述（由申请人提供）：肾结石的遗传成分是一种复杂的多基因性状，这是一个难以解开的巨大挑战。对肾结石单基因病因的研究对于揭示肾结石的遗传成分提供了最丰富的信息。钙质结石常见于先天性远端肾小管酸中毒患者：dRTA；但 sd RcoTnAs 认为钙结石是一种非常罕见的原因，因为这种疾病的患病率较低。先天性 dRTA 是一种肾酸化疾病，由液泡 :VPa;se-A T :B1 或 a4 亚基、阴离子交换器 1 或碳酸酐酶 II 突变引起。该提案重点关注 V-ATP 酶的 B1 亚基。人们普遍认为杂合携带者状态是沉默的。 B1 亚基突变导致泵功能障碍的机制尚不清楚。我们将在该提案中检验 3 个假设： I. B1 杂合突变在结石形成人群中并不罕见。二.杂合性的负担并不是沉默的，而是赋予磷酸钙或草酸盐肾结石的倾向或遭受其折磨。三．突变体 B1 亚基通过多种机制导致泵功能障碍，例如泵组装缺陷、ATP 结合/水解受损以及与 HE 易位的耦合紊乱。我们将通过 3 个目的来检验这些假设： 1. 基于人群的研究：定义结石形成和非结石人群中 B1 中所有碱基变化的等位基因频率，并收集这些个体相应的临床数据。 2. 住院人体研究：对选定的 B1 杂合个体进行详细的代谢和理化研究，以表征其表型。 3. 基准研究：利用细胞生物学和生物化学定义各种突变 B1 亚基如何导致 HE 泵功能障碍的机制。这不是对经典 dRTA 的研究。相反，我们也许能够证明单基因倾向实际上在一般结石形成人群中并不罕见，并且许多结石形成者可能含有 B1 突变，使他们容易患钙结石。这可能为一般结石形成人群中的其他单基因倾向设定优先权。这些研究还将显着增进对 V-ATP 酶生物学的理解。这些自然点突变及其同源功能障碍模式将为 B1 蛋白的生物学提供有价值的结构功能信息。公共健康相关性：每个人一生中有 10-20% 的机会患上肾结石。如果一个人的家庭成员患有肾结石，那么该人患肾结石的风险就会更高。这个过程可能涉及许多基因，但在某些情况下，单个基因可能会使人面临肾结石的风险，例如远端肾小管酸中毒（缩写为 dRTA）。大多数 dRTA 患者会出现钙肾结石，但大多数钙肾结石患者不会出现 dRTA。该提案审查了一些新想法。根据我们去年的工作，我们认为一种较温和的 dRTA 形式（即遗传一个而不是两个缺陷基因）（称为“杂合状态”）在钙肾结石形成者的一般人群中可能相当常见。该基因编码一种名为“B1”的蛋白质，该蛋白质在异常状态下会改变尿液中的成分，导致肾结石形成。我们将利用人群和实验室研究来回答三个问题。 1. 钙肾结石患者中杂合状态的异常 B1 到底有多常见？ 2. 如果一个人携带异常的 B1 拷贝（杂合状态），那么尿液中的异常到底是什么导致该人容易患肾结石？ 3. B1 异常如何改变我们尿液的成分从而形成肾结石？这些问题得到解答后，可以帮助我们了解肾结石的遗传成分，以及 B1 等单一蛋白质的异常如何以不利的方式改变我们的尿液成分，从而使我们能够更好地预测、诊断和治疗肾结石。