Mouse Model for Diseases of Protein Misfolding

蛋白质错误折叠疾病的小鼠模型

• 批准号: 8805919
• 负责人: P. MICHAEL CONN
• 金额: $ 3.87万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-07 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8805919
• 关键词: Mouse; Model; Diseases; Protein; Misfolding

Project Summary/Abstract: This project is designed to develop and characterize two prototypic genetically modified mouse models for human diseases of protein folding. These models are required to bring a novel class of target-specific drugs, pharmacoperones, to human (and animal) use. Misfolded mutant proteins are detected by the cellular quality control system (QCS) and are typically retained in the endoplasmic reticulum (ER) for either reprocessing or degradation; frequently, these mutants result in disease. Studies in cell cultures indicate that these mutants can be rescued by target-specific small molecules (pharmacoperones) which enter cells, serve as templates that refold the mutants, and permit their passage to the plasma membrane. Many mutants retain or regain their fundamental properties as ion channels, enzymes or receptors when re-routed correctly. Diseases caused by misfolding (which may benefit from this approach) include cystic fibrosis, hypogonadotropic hypogonadism, nephrogenic diabetes insipidus, retinitis pigmentosa, hypercholesterolemia, cataracts, neurodegenerative diseases (Huntington's, Alzheimer's and Parkinson's), cancers and digestive disorders. It is fair to say that virtually every person will be affected by protein folding diseases during his or her lifetime, either directly or due to the illness of a loved one. In spite of this, there are few model systems, and none in small laboratory animals, that allow the translation of available in vitro data or the testing of "hits" from high throughput screening on protein rescue into in vivo systems. To date, therapeutic approaches in humans have relied on a small number of studies in end-of-life patients, using drugs that have never been fully characterized in animal models. Such models are needed to address drug safety, the pattern of drug administration required to optimize therapeutic effectiveness, and serve as a test model for new drugs in preclinical studies. The pattern (route, dose and frequency) is particularly important as the persistence of these drugs frequently inhibits the desired activity, once rescue has occurred, so they must be removed. Moreover a convenient laboratory model for these diseases is needed if the use of pharmacoperones is to translate to human well-being, since US law requires animal testing prior to normal human volunteers. The present study will characterize prototypic mouse models of misfolding, relying on an unusually well-characterized mutant of a physiologically important GPCR (i.e. the gonadotropin releasing hormone receptor). There is much information available on the mechanism of activation of the gonadotropin releasing hormone receptor (GnRHR) and on the biochemical mechanism by which the mutant E90K is believed to cause the disease state. This information has been helpful in guiding our choices of mutant E90K and will contribute to the success of this project. The two models to be used are available and have the predicted genotype and necessary phenotype.

项目摘要/摘要： 该项目旨在开发和表征两种原型转基因小鼠模型 人类蛋白质折叠疾病。这些模型需要带来一类新型的靶向药物， 药用哌酮，供人类（和动物）使用。通过细胞质量检测错误折叠的突变蛋白 控制系统（QCS），通常保留在内质网（ER）中用于再处理或 降解;通常，这些突变体会导致疾病。细胞培养研究表明这些突变体 可以通过进入细胞并充当模板的特定目标小分子（pharmacoperones）来拯救 重新折叠突变体，并允许它们进入质膜。许多变种人保留或恢复了他们的 当正确重新路由时，离子通道、酶或受体的基本特性。引起的疾病 错误折叠（可能受益于这种方法）包括囊性纤维化、低促性腺素性性腺功能减退症、 肾性尿崩症、色素性视网膜炎、高胆固醇血症、白内障、神经退行性疾病 疾病（亨廷顿病、阿尔茨海默病和帕金森病）、癌症和消化系统疾病。可以公平地说 几乎每个人在其一生中都会受到蛋白质折叠疾病的影响，无论是直接的还是由于 为所爱的人生病。尽管如此，模型系统还很少，而且在小型实验室中也没有。 动物，允许翻译可用的体外数据或测试高通量的“命中” 筛选体内系统中的蛋白质救援。迄今为止，人类的治疗方法依赖于 对临终患者进行的少量研究，使用的药物从未在动物身上得到充分表征 模型。需要这样的模型来解决药物安全问题，优化药物管理模式 治疗效果，并作为新药临床前研究的测试模型。模式（路线， 剂量和频率）尤其重要，因为这些药物的持久性经常会抑制所需的效果 一旦发生救援，就必须将其移除。此外，还有一个方便的实验室模型 如果要将药用哌酮的使用转化为人类福祉，这些疾病是必要的，因为美国法律 需要在正常人类志愿者之前进行动物测试。本研究将表征原型 错误折叠的小鼠模型，依赖于生理上重要的一个异常良好表征的突变体 GPCR（即促性腺激素释放激素受体）。网上有很多信息可供参考 促性腺激素释放激素受体（GnRHR）的激活机制及其对生化的影响 突变体 E90K 被认为是导致疾病状态的机制。这些信息很有帮助 指导我们选择突变体 E90K，并将有助于该项目的成功。两个模型要 使用的基因是可用的并且具有预测的基因型和必要的表型。