Mouse Models with Regulatable Cell Type-specific Expression of Anti-Tau shRNAs

具有可调节细胞类型特异性抗 Tau shRNA 表达的小鼠模型

• 批准号: 8064626
• 负责人: Lennart Mucke
• 金额: $ 27.7万
• 依托单位: J. DAVID GLADSTONE INSTITUTES
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8064626
• 关键词: Abbreviations; Address; Adult; Adverse effects; Alzheimer' s Disease; Amyloid; Amyloid beta-Protein Precursor; Autopsy; Biochemical; Biological Neural Networks; Brain; Brain region; Butyric Acids; Calcium; Calmodulin; Corpus striatum structure; Data; Dependovirus; Deposition; Development; Disease; Electroencephalogram; Electroencephalography; Employee Strikes; EphB2 Receptor; Epilepsy; Excitatory Postsynaptic Potentials; Frontotemporal Dementia; Generations; Genes; Glial Fibrillary Acidic Protein; Goals; Green Fluorescent Proteins; Growth; Human; Impaired cognition; Impairment; Knowledge; Lead; Learning; Long-Term Potentiation; Measures; Memory; Memory impairment; Microtubule-Associated Proteins; Modeling; Molecular; Mus; Nature; Neurofibrillary Tangles; Neurologic; Neuronal Dysfunction; Neurons; Pathogenesis; Peptides; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Physiological; Platelet-Derived Growth Factor; Play; Proteins; RNA Interference; Regulation; Resistance; Role; Safety; Seizures; Small Interfering RNA; Solubility; Synapses; Tauopathies; Tetracycline Control; Therapeutic; Trans-Activators; Transgenic Mice; Transgenic Model; Transgenic Organisms; Viral Vector; cell type; dentate gyrus; enhanced green fluorescent protein; entorhinal cortex; excitotoxicity; gamma-Aminobutyric Acid; inhibitory neuron; innovation; mature animal; mouse model; nervous system disorder; neuropeptide Y; novel; novel therapeutics; pre-clinical; prevent; research study; small hairpin RNA; tau Proteins; tau expression; tau function; tau mutation; vesicular GABA transporter

DESCRIPTION (provided by applicant): Tau, a major microtubule-associated protein (MAP), has been implicated in Alzheimer's disease (AD) and a variety of other neurological conditions collectively referred to as tauopathies. Remarkably, the functions tau fulfills in the adult brain and the exact roles it plays in the pathogenesis of these conditions remain uncertain. The current application aims to address these flagrant knowledge gaps by generating novel mouse models in which the expression of tau can be suppressed in specific brain regions and neural cell types in adult mice. Another important factor involved in AD is the amyloid-¿ (A¿) peptide, which is derived from the amyloid precursor protein (APP). Crossing human APP (hAPP) transgenic (TG) mice with high levels of A¿ in the brain onto a complete (Tau-/-) or partial (Tau+/-) tau-deficient background prevented most of their A¿-dependent abnormalities, including impairments in learning and memory, abnormal neural network activity, and various related biochemical and anatomical neuronal alterations. Tau reduction achieved this striking rescue without changing A¿ levels or deposition in the brain. Furthermore, our hAPP/Tau+/+ mice never develop neurofibrillary tangles, and their wildtype murine tau is expressed at physiological levels, has normal solubility, and shows no evidence for the type of abnormal phosphorylation or aggregation seen in AD and mutant-tau TG models. Tau reduction is the most effective strategy to prevent A¿-induced neuronal deficits we have identified so far, but the safety of this approach in adult animals and the underlying mechanisms remain unknown. Our preliminary studies also revealed that tau reduction makes mice without hAPP/A¿ expression more resistant against chemically induced seizures, suggesting a novel role for tau in the regulation of neuronal/synaptic activity. However, why and how tau reduction increases resistance to epileptic seizures is unknown. The above constellation of preliminary findings and unresolved questions underlines the significance and novelty of the experiments described in the current application. We currently have no effective ways to treat or prevent AD. If tau reduction was as efficacious and safe in preventing cognitive dysfunction in humans with AD as it is in hAPP mice, further exploration of this strategy might lead to a therapeutic breakthrough. In addition, tau reduction might be of benefit in other neurological disorders in which excitotoxicity plays a role. To address these challenges and opportunities, we propose the following specific aims. Aim 1. Decrease tau levels in brains of adult hAPP mice and NTG controls with viral vectors expressing anti-tau shRNA. Aim 2. Generate transgenic mice with regulatable expression of anti-tau shRNA. Aim 3. Analyze the models generated in Aims 1 and 2 behaviorally, anatomically and biochemically. Generation of the proposed mouse models should allow us to directly address the following questions to which there currently are no clear answers: Is tau reduction efficacious and safe when implemented in adulthood? In which brain region and neuronal cell types (e.g., neuronal vs. glial, excitatory vs. inhibitory neurons) does tau have to be reduced in order to prevent A¿- dependent cognitive decline and to increase resistance against chemically induced seizures? In the long run, the proposed models could help decipher the function(s) of tau in the adult brain, its role in disease-related neuronal dysfunction, and the underlying molecular mechanisms. PUBLIC HEALTH RELEVANCE: Amyloid-¿ (A¿) is widely thought to cause Alzheimer's disease (AD). We recently discovered that reducing the protein Tau can prevent A¿ from causing memory deficits and related neuronal impairments in AD mouse models. In this proposal, we will assess the efficacy and safety of this novel therapeutic strategy at the preclinical level.

描述（由应用程序提供）：Tau是一种主要的微管相关蛋白（MAP），在阿尔茨海默氏病（AD）中实施了，以及多种其他神经系统疾病，共同称为tauopathies。值得注意的是，tau在成人大脑中的功能及其在这些疾病的发病机理中起的确切作用仍然不确定。当前的应用旨在通过产生新的小鼠模型来解决这些公然的知识差距，在成年小鼠的特定大脑区域和神经细胞类型中可以抑制Tau的表达。 AD中涉及的另一个重要因素是淀粉样蛋白（A¿）肽，该肽是从淀粉样蛋白前体蛋白（APP）得出的。跨越大脑中高水平A的人类应用（HAPP）转基因（TG）小鼠，将其跨到完整的（TAU - / - ）或部分（TAU +/-）tau-Deeigent背景，阻止了它们的大部分依赖性异常，包括学习和记忆中的损害，包括学习和记忆的损害，异常的神经神经元网络活动，以及各种相关的bioChemical and Anatal和各种相关的nuaron和各种关系。 tau减少了这次罢工，而没有改变大脑中的A水平或沉积。此外，我们的Happ/tau+/+小鼠从未产生过神经原纤维缠结，它们的野生型鼠tau在物理水平上表达，具有正常的溶解度，并且没有证据表明在AD和Mutant-Tau TG模型中看到的异常磷酸化或聚集的类型。减少tau是防止A诱导的神经元定义我们迄今已确定的最有效的策略，但是这种方法在成年动物和基本机制中的安全性仍然未知。我们的初步研究还表明，tau的减少使小鼠没有happ/a表达对化学诱导的癫痫发作具有更具耐药性，这表明tau在调节神经元/突触活性中起着新的作用。然而，为什么和如何减少tau增加对癫痫发作的抗性尚不清楚。上述初步发现和未解决的问题的星座强调了当前应用中描述的实验的重要性和新颖性。我们目前没有有效的方法来治疗或预防广告。如果减少tau在预防AD的认知功能障碍方面与HAPP小鼠的认知功能障碍一样有效，安全，那么对这种策略的进一步探索可能会导致治疗性突破。此外，在兴奋性毒性起作用的其他神经系统疾病中，tau降低可能有益。为了应对这些挑战和机遇，我们提出以下特定目标。 AIM 1。使用表达抗Tau shRNA的病毒向量降低成年Happ小鼠和NTG对照的大脑中的tau水平。 AIM 2。产生具有抗Tau shRNA的调节表达的转基因小鼠。目标3。分析目标1和2中在行为，解剖和生化上产生的模型。所提出的鼠标模型的产生应该使我们能够直接解决当前没有明确答案的以下问题：成年后实施时tau是否有效且安全？在哪种脑区域和神经元细胞类型中（例如，神经元与神经胶质，兴奋性与抑制性神经元）是否必须减少tau，以防止A-依赖性认知下降并增加对化学诱导癫痫发作的抵抗力？从长远来看，提出的模型可以帮助破译成人大脑中Tau的功能，其在与疾病相关的神经元功能障碍中的作用以及基本的分子机制。 公共卫生相关性：淀粉样蛋白（a。）被普遍认为会引起阿尔茨海默氏病（AD）。我们最近发现，减少蛋白质TAU可以防止A在AD小鼠模型中引起记忆定义和相关的神经元损伤。在此提案中，我们将评估临床前这种新型治疗策略的有效性和安全性。