Mouse Model for Diseases of Protein Misfolding

蛋白质错误折叠疾病的小鼠模型

基本信息

批准号：
8033595
负责人：
P. MICHAEL CONN
金额：
$ 26.25万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-01-01 至 2012-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8033595
关键词：
Address Affect Agonist Alzheimer&apos s Disease Anatomy Animal Diseases Animal Model Animal Testing Animals Biochemical Biological Assay Biological Models Breeding Cataract Cell Culture System Cell Culture Techniques Cell membrane Cells Cystic Fibrosis Data Development Digestive System Disorders Disease Disease model Dose Drug usage Endoplasmic Reticulum Enzymes Frequencies Funding G Protein-Coupled Receptor Genes Gene Targeting Genetic Genetically Modified Animals Genotype Gonadotropin-Releasing Hormone Receptor Grant Histology Hormonal Change Human Human Volunteers Huntington Disease Hypogonadism In Vitro Ion Channel Klinefelter&apos s Syndrome Laboratories Laboratory Animals Laws Malignant Neoplasms Maps Modeling Molecular Chaperones Monitor Mus Mutation Nephrogenic Diabetes Insipidus Neurodegenerative Disorders Parkinson Disease Patients Pattern Personal Satisfaction Persons Pharmaceutical Preparations Phenotype Pituitary Gland Production Property Protein-Folding Disease Proteins Protocols documentation Quality Control Reagent Retinitis Pigmentosa Route Safety Site System Technical Expertise Testing Therapeutic Tissues Translating Translations Treatment Efficacy Validation Work base design end of life high throughput screening human disease hypercholesterolemia in vivo in vivo Model loved ones mouse model mutant novel offspring peptidomimetics preclinical study protein folding protein misfolding prototype receptor small molecule small molecule libraries success therapeutic effectiveness

项目摘要

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: This project is designed to develop, characterize, compare and contrast two models for human diseases of protein folding. Diseases caused by misfolding include cystic fibrosis, hypogonadotropic hypogonadism, nephrogenic diabetes insipidus, retinitis pigmentosa, hypercholesterolemia, cataracts, neurodegenerative diseases (Huntington's, Alzheimer's and Parkinson's), particular cancers and a number of digestive disorders resulting from enzyme mutation. 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.

描述（由申请人提供）：该项目旨在开发和表征两个原型遗传修饰的小鼠模型，用于蛋白质折叠的人类疾病。这些模型需要将新型的针对特定目标药物（药物）带入人类（和动物）的使用。通过细胞质量控制系统（QC）检测到错误折叠的突变蛋白，通常保留在内质网（ER）中以进行后处理或降解。这些突变体经常会导致疾病。细胞培养物的研究表明，这些突变体可以通过进入细胞的靶标特异性小分子（Pharmacoperone）来挽救这些突变体，作为重新折叠突变体的模板，并允许它们通过质膜。正确重新切换后，许多突变体将其作为离子通道，酶或受体保留或恢复其基本特性。因折叠率错误（可能从这种方法中受益）引起的疾病包括囊性纤维化，性核腺功能减退，肾脏糖糖尿病，色素性阴影性色素，高胆固醇血症，cataracts，cataracts，神经退行性疾病（神经退行性疾病）（亨廷顿的Alzheimer和Parkisers和Parkinson和Parkinsons）和Parkinsons和Parkinsons，可以公平地说，几乎每个人都会直接或由于亲人的疾病而受到蛋白质折叠疾病的影响。尽管如此，在小型实验室动物中，几乎没有模型系统，允许可用的体外数据或从蛋白质救援对体内系统的高吞吐量筛选中进行“命中”的测试。迄今为止，使用从未在动物模型中从未充分表征的药物，人类的治疗方法依赖于寿命末患者的少量研究。需要这样的模型来解决药物安全，这是优化治疗有效性所需的药物管理模式，并作为临床前研究中新药的测试模型。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抑制所需的活动，一旦发生救援，就必须将其去除。此外，如果使用Pharmacoperones是将人类福祉转化为人类福祉，则需要方便这些疾病的实验室模型，因为美国法律需要在正常的人类志愿者之前进行动物测试。本研究将依靠生理上重要的GPCR（即促性腺激素释放激素受体）的异常表征的突变体来表征错误折叠的原型小鼠模型。关于促性腺激素释放激素受体（GNRHR）的激活机制以及生化机制的激活机制有很多信息。这些信息有助于指导我们对突变E90K的选择，并将有助于该项目的成功。要使用的两个模型可用，并具有预测的基因型和必要的表型。公共卫生相关性：该项目旨在开发，表征，比较和对比蛋白质折叠的两个模型。由误折折叠引起的疾病包括囊性纤维化，性肾上腺素性性腺功能不全，肾脏糖尿病性肠道疾病，色素性视网膜炎，高胆固醇血症，白内障，神经退行性疾病（Huntington，Alzheimer's and Alzheimer's和Parkinson's），特定的犬类和数字疾病，疾病和数字是数字。突变。如果使用Pharmacoperones是转化为人类福祉，则需要用于这些疾病的方便实验室模型，因为美国法律需要在正常的人类志愿者之前进行动物测试。