Antisense Oligonucleotides targeting APP to prevent neurodegeneration in models of Down Syndrome and Alzheimer's disease

靶向 APP 的反义寡核苷酸可预防唐氏综合症和阿尔茨海默病模型中的神经变性

• 批准号: 10092057
• 负责人: William C Mobley
• 金额: $ 73.21万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-15 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10092057
• 关键词: Age; Alzheimer' s Disease; Alzheimer' s disease model; Amyloid; Amyloid beta-42; Amyloid beta-Protein Precursor; Antisense Oligonucleotides; Atrophic; Benchmarking; Blinded; C-terminal; Central Nervous System Diseases; Chromosome 21; Code; Cognition; Cognitive deficits; Collaborations; Complex; Dementia; Deposition; Disease; Dose; Down Syndrome; Drug Kinetics; Early Endosome; Elderly; Endosomes; Gene Expression; Gene Proteins; Genes; Genome; Goals; Guanosine Triphosphate Phosphohydrolases; Human Amyloid Precursor Protein; In Vitro; Injections; Intracellular Transport; Length; Measures; Mediating; Messenger RNA; Modeling; Mus; Nerve Degeneration; Neurofibrillary Tangles; Neurons; Outcome; Pharmacodynamics; Phenotype; Population; Property; Proteins; RNA; RTN4 gene; Sampling; Senile Plaques; Signal Transduction; Specificity; Spinal Muscular Atrophy; Structure; Synapses; System; Testing; Therapeutic; Tissues; Toxic effect; Transgenic Organisms; Treatment Efficacy; Treatment Protocols; Trisomy; Vertebral column; age related neurodegeneration; basal forebrain; base; cerebral amyloidosis; cholinergic; design; entorhinal cortex; improved; in vivo; inorganic phosphate; locus ceruleus structure; mouse model; mutant mouse model; neuropathology; neurotrophic factor; pharmacokinetics and pharmacodynamics; phosphorothioate; prevent; protein expression; tau Proteins; tau-1; therapeutic evaluation; trafficking

We aim to prevent Alzheimer disease (AD) in DS (trisomy 21) (AD-DS). Using antisense oligonucleotides (ASOs), we will selectively target RNA for the amyloid precursor protein (APP) in mouse models of AD-DS (Dp16) and AD/cerebral amyloidosis (Line 41). The therapeutic premise is based on: 1) increased APP gene dose is necessary for AD-DS. As replicated in models of AD-DS, normalizing APP dose eliminated: a) age-related neurodegeneration in locus coeruleus and the basal forebrain complex, b) hyper- phosphorylation of Tau, and c) enlargement of early endosomes; 2) pointing to a mechanism by which increased APP gene dose acts, increased full-length APP (fl-APP), its 99 residue C-terminal fragment (C99) and Aβ42 each increased Rab5 activity, thus enlarging early endosomes, disrupting endosomal trafficking of neurotrophic signals, and causing atrophy of BFCNs; 3) therefore, reducing levels of these APP products is a rational approach to preventing or lessening the impact of increased APP gene dose in AD-DS, including effects on endosomes. ASOs have recently been shown to safely and effectively treat CNS disorders. Indeed, FDA approval for ASOs in Spinal Muscular Atrophy motivates trials of ASOs in other CNS diseases. In preliminary studies we showed that intracerebroventricular (ICV) injection of ASOs targeting mouse and human APP (i.e. mAPP-ASOs and hAPP-ASOs) reduced APP mRNA and protein levels. Using mouse models of AD-DS and AD/cerebral amyloidosis we will test the therapeutic hypothesis that APP-ASOs will selectively reduce the levels of APP mRNA and its products to prevent and/or lessen neurodegeneration. The mechanistic hypothesis is that APP-ASOs will normalize endosomal structure and function, neurotrophin signaling and trafficking, and improve cognition. Using defined GO/NOGO criteria as a guide, we will pursue these Specific Aims: 1. To investigate newly designed APP-ASOs in vitro for efficacy and target specificity. Using an existing mAPP-ASO as benchmark, additional mAPP-ASOs will be designed to increase potency for targeting APP mRNA and its products and normalizing endosome size. 2. To establish optimal APP-ASO doses and dose-intervals based on empirically defined in vivo pharmacokinetic (PK) and pharmacodynamic (PD) properties. We will define effective, non-toxic doses and treatment intervals for advancement of mAPP-ASOs and hAPP-ASOs to in vivo studies in Aim 3. In the Dp16 model, we will target a ~33% reduction of mAPP RNA, i.e. to 2N values; in Line 41 mice we will target a 50% reduction. Aim 3. To investigate in vivo APP-ASO efficacy in ameliorating neurodegeneration and normalizing endosomal phenotypes. To test the therapeutic hypothesis, we will ask if APP-ASOs given before degeneration in Dp16 mice and plaque deposition in Line 41 mice prevent these changes. Next, we will ask if degeneration in Dp16 mice can be reversed by APP-ASO treatment. The mechanistic hypothesis will be informed by whether or not APP-ASO reductions in degeneration are correlated with normalization of endosomal phenotypes.

我们旨在预防DS（三体疾病）（AD-DS）中的阿尔茨海默氏病（AD）。使用反义寡核苷酸 （ASOS），我们将选择性地靶向AD-DS小鼠模型中淀粉样蛋白前体蛋白（APP）的RNA（DP16） 和AD/脑淀粉样变性（第41行）。理论前提是基于：1）增加的应用基因剂量是 AD-DS所需的。如在AD-DS模型中复制的，消除了应用程序剂量：a）与年龄有关 基因座和基本前脑复合物中的神经变性，b）tau的高磷酸化，c） 早期内体的扩展； 2）指出一种机制，通过该机制增加了APP基因剂量作用，全长增加 APP（FL-APP），其99居住C末端碎片（C99）和Aβ42各个rab5活性，从而提早增加 内体，破坏神经营养信号的内体运输，并导致BFCN萎缩； 3） 因此，降低这些应用产品的水平是防止或减轻影响的合理方法 AD-DS中的APP基因剂量增加，包括对内体的影响。最近已显示ASOS安全 并有效治疗中枢神经系统疾病。实际上，FDA在脊柱肌肉萎缩中批准ASO会激发试验 其他中枢神经系统疾病中的ASO。在初步研究中，我们表明脑室内（ICV）注射 ASOS靶向小鼠和人类应用程序（即mapp-asos和happ-asos）降低了应用mRNA和蛋白质 水平。使用AD-DS和AD/脑淀粉样变性的小鼠模型，我们将测试治疗假设 该应用程序将有选择地降低App mRNA及其产品的水平，以预防和/或减少 神经变性。机械假设是App-AsoS将正常化内体结构和 功能，神经营养蛋白信号传导和运输以及改善认知。使用定义的go/nogo标准为 指南，我们将追求以下具体目标：1。调查在体外设计的新设计的App-asos，以提高效率和 目标特异性。使用现有的Mapp-Aso作为基准，将设计其他Mapp-Asos 提高靶向应用程序mRNA及其产品的效力并使内体大小正常化。 2。建立 基于体内药代动力学（PK）和 药学（PD）特性。我们将定义有效的，无毒的剂量和治疗间隔 Mapp-asos和Happ-asos在AIM 3中进行体内研究的进步。在DP16模型中，我们将针对A 〜33％的MAPP RNA降低，即2N值；在第41小鼠中，我们将降低50％。目标3 研究体内APP-ASO效率在改善神经变性和归一化内体 表型。为了检验治疗假设，我们将询问DP16退化之前给出的APP-ASO是否给出 小鼠和斑块沉积在第41列小鼠中阻止了这些变化。接下来，我们将询问DP16中的退化 小鼠可以通过APP-ASO治疗逆转。机械假设将通过是否 变性的APP-ASO减少与内体表型的归一化相关。