Generating iPSC-derived Neurons to Explore Formation & Inhibition of Human Prions

生成 iPSC 衍生神经元以探索形成

• 批准号: 8824349
• 负责人: XIN QI
• 金额: $ 23.78万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-30 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8824349
• 关键词: Age; Alzheimer' s Disease; Animal Model; Animals; Cell Line; Cell model; Cells; Cessation of life; Clinical Trials; Creutzfeldt-Jakob Syndrome; Culture Media; Derivation procedure; Detergents; Disease; Event; Fibroblasts; Genetic; Goals; Health; Human; In Vitro; Individual; Infection; Laboratories; Lead; Life; Link; Modeling; Molecular; Monitor; Mus; Mutation; Nerve Degeneration; Neuraxis; Neurites; Neurodegenerative Disorders; Neurons; Parkinson Disease; Pathogenesis; Pathway interactions; Patients; Peptide Hydrolases; Pharmaceutical Preparations; Phenotype; Pluripotent Stem Cells; PrP; PrPSc Proteins; Prion Diseases; Prions; Process; Proteins; Recombinants; Research; Resistance; Resources; Role; Sampling; Skin; Solubility; Syndrome; System; Testing; Therapeutic; Work; abnormal PrP; base; cell type; conformational conversion; disease phenotype; human PrP; improved; in vivo; induced pluripotent stem cell; inhibitor/antagonist; insight; mutant; mutation carrier; nerve stem cell; neurotoxicity; novel; pathogen; prevent; prion-like; protein aggregate; protein misfolding; response; trafficking; wild-type PrP

DESCRIPTION (provided by applicant): The key molecular mechanism of the currently incurable human prion diseases, a group of transmissible neurodegenerative disorders, involves prion formation due to a conformational transition from the cellular prion protein (PrPC) into its pathological form (PrPSc) in the central nervous system. Present cell and animal models do not seem to work for human prion diseases well since potential anti-prion compounds identified using these models failed in clinical trials. Clearly, the lack of appropriate models that are able to faithfully mimic in vivo human PrPC to PrPSc conversion and prion-associated neurotoxicity not only significantly limits our understanding of the molecular mechanism of the conversion but also confines developing of therapeutic drugs. The challenges may be overcome by obtaining various live human neurons using the newly-developed approach by reprogramming patient-derived fibroblasts into induced pluripotent stem cells (iPSCs) (Takahashi et al., 2007). Using skin-derived iPSCs from asymptomatic subjects carrying PrP mutation such as E200K, D178N, F198S, or a new mutation E200G, linked to genetic prion disease recently generated in our laboratory, we propose to further differentiate the patient-specific iPSCs into neurons to test our hypothesis that patient- specific iPSC-derived neurons can be used as models for monitoring disease phenotypes and developing therapeutic strategies for prion diseases. In this application, we propose to differentiate patient- specific iPSCs into neurons and to characterize the molecular identity of differentiated cells. Then, the patient- specific iPSC-derived neurons generated in this study will be used to characterize disease-associated phenotypes and to investigate cellular and molecular mechanisms of neurodegeneration induced by human prions and protective roles of recombinant human PrP and GSK2606414, a new specific inhibitor of a pathway of unfolded protein response. We believe that successful implementation of our study will not only generate patient-specific iPSC-derived neurons but also provide insights into the pathogenesis and treatment of prion diseases. Moreover, our study may be significant in improving our understanding of other neurodegenerative disorders such as Alzheimer's and Parkinson's disease that may involve a prion-like pathogenic mechanism as well.

描述（申请人提供）：目前无法治愈的人类朊病毒疾病是一组传染性神经退行性疾病，其关键分子机制涉及由于中枢神经系统中细胞朊病毒蛋白（PrPC）构象转变为其病理形式（PrPSc）而形成朊病毒。目前的细胞和动物模型似乎对人类朊病毒疾病效果不佳，因为使用这些模型鉴定的潜在抗朊病毒化合物在临床试验中失败了。显然，缺乏合适的模型能够 忠实地模拟体内人 PrPC 到 PrPSc 的转化和朊病毒相关的神经毒性不仅极大地限制了我们对转化分子机制的理解，而且限制了治疗药物的开发。通过使用新开发的方法将患者来源的成纤维细胞重编程为诱导多能干细胞（iPSC）来获得各种活的人类神经元，可以克服这些挑战（Takahashi et al., 2007）。使用来自携带 PrP 突变（例如 E200K、D178N、F198S 或新突变 E200G（与我们实验室最近产生的遗传性朊病毒病相关）的无症状受试者的皮肤来源的 iPSC，我们建议进一步将患者特异性 iPSC 分化为神经元，以测试我们的研究结果。 假设患者特异性 iPSC 衍生神经元可用作监测疾病表型和制定朊病毒疾病治疗策略的模型。在此应用中，我们建议将患者特异性 iPSC 分化为神经元，并表征分化细胞的分子身份。然后，本研究中产生的患者特异性 iPSC 衍生神经元将用于表征疾病相关表型，并研究人朊病毒诱导的神经变性的细胞和分子机制以及重组人 PrP 和 GSK2606414（一种未折叠蛋白反应途径的新型特异性抑制剂）的保护作用。我们相信，我们研究的成功实施不仅将产生患者特异性 iPSC 衍生神经元，而且还将为朊病毒疾病的发病机制和治疗提供见解。此外，我们的研究对于提高我们对其他神经退行性疾病（例如阿尔茨海默病和帕金森病）的理解可能具有重要意义，这些疾病也可能涉及类似朊病毒的致病机制。