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: 10554249
• 负责人: William C Mobley
• 金额: $ 64.25万
• 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 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10554249
• 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; Life; Maintenance; 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; cerebral amyloidosis; cholinergic; design; entorhinal cortex; hyperphosphorylated tau; 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.

我们的目标是预防 DS（21 三体）(AD-DS) 患者的阿尔茨海默病 (AD)。使用反义寡核苷酸 (ASO)，我们将在 AD-DS (Dp16) 小鼠模型中选择性靶向淀粉样前体蛋白 (APP) 的 RNA 和 AD/脑淀粉样变性（第 41 行）。治疗前提基于：1）增加APP基因剂量为 AD-DS 所必需的。正如 AD-DS 模型中所复制的那样，标准化 APP 剂量消除了：a) 与年龄相关的 蓝斑和基底前脑复合体的神经变性，b) Tau 过度磷酸化，c) 早期内体增大； 2)指出增加APP基因剂量作用、增加全长的机制 APP (fl-APP)、其 99 个残基 C 端片段 (C99) 和 Aβ42 均增加 Rab5 活性，从而早期放大 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 APP 产品的含量是预防或减轻影响的合理方法。 AD-DS 中 APP 基因剂量增加，包括对内涵体的影响。 ASO 最近被证明可以安全地 并有效治疗中枢神经系统疾病。事实上，FDA 批准 ASO 治疗脊髓性肌萎缩症激发了试验 ASOs 在其他中枢神经系统疾病中的作用。在初步研究中，我们表明，脑室内（ICV）注射 靶向小鼠和人类 APP 的 ASO（即 mAPP-ASO 和 hAPP-ASO）可减少 APP mRNA 和蛋白质 水平。我们将使用 AD-DS 和 AD/脑淀粉样变性小鼠模型来检验治疗假设 APP-ASO 将选择性地降低 APP mRNA 及其产物的水平，以预防和/或减轻 神经变性。机制假设是 APP-ASO 将使内体结构正常化并 功能、神经营养素信号传导和运输，并改善认知。使用定义的 GO/NOGO 标准作为 作为指南，我们将追求以下具体目标： 1. 体外研究新设计的 APP-ASO 的功效和 目标特异性。使用现有的 mAPP-ASO 作为基准，将设计额外的 mAPP-ASO 增加靶向 APP mRNA 及其产物并使内体大小正常化的效力。 2. 建立 最佳 APP-ASO 剂量和剂量间隔基于经验定义的体内药代动力学 (PK) 和 药效学（PD）特性。我们将确定有效、无毒的剂量和治疗间隔 将 mAPP-ASO 和 hAPP-ASO 推进目标 3 中的体内研究。在 Dp16 模型中，我们将目标 mAPP RNA 减少约 33%，即减少至 2N 值；在 41 系小鼠中，我们的目标是减少 50%。目标 3. 至 研究体内 APP-ASO 在改善神经退行性变和使内体正常化方面的功效 表型。为了检验治疗假设，我们将询问是否在 Dp16 变性之前给予 APP-ASO 小鼠和 41 系小鼠中的斑块沉积阻止了这些变化。接下来我们要问Dp16是否退化 小鼠可以通过 APP-ASO 治疗来逆转。机械假说将取决于是否 APP-ASO 变性的减少与内体表型的正常化相关。

项目成果

Prenatal and Postnatal Pharmacotherapy in Down Syndrome: The Search to Prevent or Ameliorate Neurodevelopmental and Neurodegenerative Disorders.

• DOI: 10.1146/annurev-pharmtox-041521-103641
• 发表时间: 2022-01-06
• 期刊: Annual review of pharmacology and toxicology
• 影响因子: 12.5

Normal levels of KIF5 but reduced KLC1 levels in both Alzheimer disease and Alzheimer disease in Down syndrome: evidence suggesting defects in anterograde transport.

• DOI: 10.1186/s13195-021-00796-6
• 发表时间: 2021-03-10
• 期刊: Alzheimer's research & therapy
• 作者: Chen XQ;Das U;Park G;Mobley WC
• 通讯作者: Mobley WC

Mechanistic Analysis of Age-Related Clinical Manifestations in Down Syndrome.

• DOI: 10.3389/fnagi.2021.700280
• 发表时间: 2021
• 期刊: Frontiers in aging neuroscience
• 影响因子: 4.8
• 作者: Chen XQ;Xing Z;Chen QD;Salvi RJ;Zhang X;Tycko B;Mobley WC;Yu YE
• 通讯作者: Yu YE