Analysis of Cytoplasmic Prion Protein Toxicity

细胞质朊病毒蛋白毒性分析

• 批准号: 6835348
• 负责人: WALKER S JACKSON
• 金额: $ 4.3万
• 依托单位: WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
• 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 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6835348
• 关键词: cell component structure /function; cell line; cytoplasm; cytotoxicity; embryonic stem cell; endoplasmic reticulum; family genetics; fibroblasts; gene expression; gene targeting; glia; intracellular transport; laboratory mouse; molecular pathology; mutant; neurons; postdoctoral investigator; prions; proteasome; protein biosynthesis; protein degradation; protein localization; protein quantitation /detection; protein structure function; spongiform encephalopathy

DESCRIPTION (provided by applicant): Evidence produced in the mid 1990's suggests that bovine spongiform encephalopathy (commonly known as mad cow disease) was probably transmitted to humans. The agents widely believed to be responsible for these diseases are abnormally shapened prion proteins. Along with an infectious route, humans can also develop prion diseases either sporadically or by expressing a mutant prion protein (penetrance approximately 100%). Recent work points to the cytosol as being the site of toxicity of prion proteins. Mutant prion proteins may be highly toxic because they have an increased probability of misfolding and therefore mislocalizing to the cytosol. To study the mechanism of increased toxicity of the mutant prion proteins and how it relates to cytosolic prion toxicity, ES cells will be genetically altered to express mutant prion proteins directly into the cytosol. These cell lines will be differentiated into neurons, gila, or fibroblasts and used to study the effects these mutations have on subcellular traffciking, degradation, and toxicity of PrP. The gene-targeting approach will allow for a high probability of equivalent expression of these constructs at the mRNA level. The selection of ES cells as a model system will permit efficient transition of cell culture experiments into mice.

描述（由申请人提供）：1990年代中期产生的证据表明，牛海绵状脑病（通常称为疯牛病）可能是传播给人类的。人们普遍认为这些疾病是造成这些疾病的造成的特工是异常塑造的prion蛋白。除了传染途径外，人类还可以通过偶尔或表达突变的prion蛋白（渗透率约100％）出现病毒疾病。最近的工作表明细胞质是prion蛋白的毒性部位。突变的prion蛋白可能是剧毒的，因为它们的折叠率错误的可能性增加，因此将其定位于细胞质溶液。为了研究突变prion蛋白毒性增加的机制及其与胞质prion毒性毒性的关系，ES细胞将在遗传上改变以直接表达突变prion蛋白直接进入细胞质。这些细胞系将分化为神经元，gila或成纤维细胞，并用于研究这些突变对PRP亚细胞贩运，降解和毒性的影响。靶向基因的方法将使这些构建体在mRNA水平上等效表达的可能性很高。 ES细胞作为模型系统的选择将允许将细胞培养实验的有效过渡到小鼠中。