Liposome-siRNA-Peptide Complexes as Therapy to Cure Prion Diseases in Mouse Model

脂质体-siRNA-肽复合物作为治疗小鼠模型中朊病毒病的疗法

• 批准号: 8616817
• 负责人: MARK D ZABEL
• 金额: $ 31.87万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-01 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8616817
• 关键词: Agriculture; Alzheimer' s Disease; Animals; Area; Behavioral; Blood - brain barrier anatomy; Bovine Spongiform Encephalopathy; Brain; Cattle; Cell Culture Techniques; Cell physiology; Cessation of life; Chronic Wasting Disease; Clinical; Cognitive; Complex; Creutzfeldt-Jakob Syndrome; Deer; Defect; Disease; Disease Progression; Early Diagnosis; Early treatment; Effectiveness; Endopeptidase K; Genetic; Human; Infection; Kinetics; Kuru; Liposomes; Methods; Modeling; Monitor; Mus; Neurodegenerative Disorders; Neurons; North America; Pathogenesis; Peptides; Personal Satisfaction; Plants; PrP; PrPC Proteins; Prion Diseases; Prions; Proteins; Public Health; RNA; RNA Interference; Regulation; Research Personnel; Resistance; Safety; Scrapie; Serum; Severity of illness; Sheep; Small Interfering RNA; Subfamily lentivirinae; System; Time; Translations; Viral; Viral Vector; Work; cancer cell; dosage; effective therapy; endoribonuclease; human DICER1 protein; improved; in vivo; knock-down; mouse model; neuropathology; novel; plasmid DNA; small hairpin RNA

DESCRIPTION (provided by applicant): Transmissible spongiform encephalopathies (TSEs) include scrapie in sheep; bovine spongiform encephalopathy (BSE) in cattle; chronic wasting disease (CWD) in deer and elk; and Kuru and Creutzfeldt-Jakob disease in humans. Accumulation of PrPRES, a proteinase K (PK) resistant form of the normal prion protein, PrPC appears central to the pathogenesis of these diseases. TSEs are invariably fatal diseases for which no early diagnosis or treatment exists. RNA interference (RNAi), a relatively new discovery in both plants and animals that regulates the amount of RNA available for translation into protein, has been used experimentally to limit the amount of viral, bacterial and cancer cell replication. Recent advances toward an effective therapy for prion diseases employ RNAi to suppress expression of a normal host protein, PrPC, in neurons and subsequent prion neuropathology, exploiting the phenomenon that disease severity and progression correlate with the amount of PrPC present in neurons. However, delivery of lentivirus encoding PrP short hairpin RNA for RNAi has demonstrated only modest effectiveness experimentally and limited use clinically due to the invasive delivery method, limited area and irreversibility of PrPC suppression and safety concerns. We have recently developed a novel small interfering RNA (siRNA) delivery system using liposome-siRNA-peptide complexes (LSPCs) to protect siRNAs from serum degradation, deliver siRNA specifically to neurons and knockdown PrPC sufficiently to cure prion disease in two cell culture models. Here we propose to extend this work to cure two distinct prion diseases in mice using LSPCs injected intravenously (iv) to deliver PrP siRNA across the blood-brain-barrier (BBB) to neurons throughout the brain. Our specific aims are as follows. SPECIFIC AIM 1: Determine kinetics of LSPC delivery and PrPC knockdown in mice to optimize LSPC therapy to treat prion disease SPECIFIC AIM 2: To determine whether PrP siRNA-containing LSPCs can cure established prion infection in mouse models of scrapie and chronic wasting disease (CWD) a. Infect mice with mouse-adapted strains of scrapie and CWD prions b. Monitor mice for early behavioral and cognitive defects prior to onset of clinical diseas c. Treat mice with PrP and control LSPCs using regiments varying in dosage and timing

描述（由申请人提供）：传染性海绵状脑病（TSEs）包括绵羊瘙痒症;牛海绵状脑病（BSE）;鹿和麋鹿慢性消耗性疾病（CWD）;以及人类库鲁病和克雅氏病。PrPRES是正常朊病毒蛋白的蛋白酶K（PK）抗性形式，PrPC的积累在这些疾病的发病机制中起重要作用。TSE总是致命的疾病，没有早期诊断或治疗。RNA干扰（RNAi）是植物和动物中一项相对较新的发现，可调节可用于翻译成蛋白质的RNA的量，已在实验中用于限制病毒，细菌和癌细胞复制的量。针对朊病毒疾病的有效治疗的最新进展利用RNAi来抑制神经元中正常宿主蛋白PrPC的表达和随后的朊病毒神经病理学，利用疾病严重程度和进展与神经元中存在的PrPC的量相关的现象。然而，用于RNAi的编码PrP短发夹RNA的慢病毒的递送在实验上仅证明了适度的有效性，并且由于侵入性递送方法、有限的区域和PrPC抑制的不可逆性以及安全性问题而限制了临床使用。我们最近开发了一种新的小干扰RNA（siRNA）传递系统，使用脂质体-siRNA-肽复合物（LSPCs），以保护siRNA免受血清降解，将siRNA特异性地传递到神经元，并在两种细胞培养模型中敲低PrPC足以治愈朊病毒疾病。在这里，我们建议将这项工作扩展到治疗两种不同的朊病毒疾病的小鼠使用静脉注射（iv）LSPC提供PrP siRNA穿过血脑屏障（BBB）的神经元在整个大脑。我们的具体目标如下。具体目标1：确定小鼠中LSPC递送和PrPC敲低的动力学以优化治疗朊病毒疾病的LSPC疗法。具体目的2：确定含有PrP siRNA的LSPC是否可以治愈羊瘙痒病和慢性消耗性疾病（CWD）的小鼠模型中建立的朊病毒感染。用鼠适应性瘙痒病和慢性消耗病朊病毒B感染小鼠。在临床疾病发作前监测小鼠的早期行为和认知缺陷。使用不同剂量和时间的方案，用PrP和对照LSPC治疗小鼠