Development of Splice Modulating Oligonucleotides Targeting RAGE as a Treatment for Alzheimer's Disease

项目总结/摘要 阿尔茨海默病（Alzheimer's Disease，AD）是一种严重的神经退行性疾病，其特征在于进行性痴呆， 记忆力减退、语言缺陷、性格改变和痴呆症，最常见于65岁以上的患者。电流 治疗对认知能力下降只有适度和暂时的影响，强调了新的治疗方法的必要性。 可以阻止AD进展的治疗。 虽然常见的AD是由淀粉样前体蛋白的常染色体显性突变引起的， 早老素，大多数AD病例被认为是散发性的，具有遗传和 环境风险因素。然而，β淀粉样蛋白（Aβ）斑块，过度磷酸化的tau神经元， 神经纤维缠结（NT）和严重的神经胶质增生都是所有形式的AD共有的神经退行性标志。受体 晚期糖基化终末产物（AGEs）在AD患者的大脑中上调， 与神经元、免疫细胞和血管细胞中的Aβ有关。信号转导还可以诱导tau蛋白的过度磷酸化 并且与散发性AD的共病风险因素（例如心血管疾病）的发病机制有关 糖尿病）。因此，AD是家族性和散发性AD治疗开发的主要靶点。 mRNA前体产生相互排斥的选择性剪接变体，其是膜- 结合并能够信号传导（mba），或截短并内源性分泌以清除mba配体 没有信号（ESSENTIAL）。在AD小鼠中，过表达mbb 3可加速AD小鼠的病理、认知 和AD的行为标志，并且输注合成的雌二醇降低了Aβ和NT病理学， 改善学习/记忆和突触功能。因此，开发了一种新的策略来减少信号传导 能够控制前体mRNA替代的剪接调节寡核苷酸（SMO）化合物， 剪接以减少mbb 3的表达并同时增加esb 3的表达。SMOs是一个 一类基于合成RNA的化合物，其在空间上阻断或减弱RNA分子之间的相互作用。 剪接机制和前体mRNA。SMOs以优异的效力、特异性、长 作用持续时间和可忽略的脱靶效应。以前靶向配体的药理学方法- 靶外效应或短半衰期阻碍了药物相互作用， 本文提出了增加天然雌激素和减少雌激素同种型表达的机制。 将通过计算机模拟设计靶向BMPs的SMOs，并针对效力和特异性进行优化，首先在体外细胞 培养（目标1），以选择领先的SMO候选药物。然后，将对每个测绘处候选人进行定性， 进一步优化了在转基因小鼠中的体内安全性和效力（Aim 2）。最后，SMOs将 作为治疗剂在急性链脲佐菌素（STZ）诱导的AD模型中评估， 认知缺陷和tau过度磷酸化（Aim 3）。最终目标是建立一个牵头的区域监测和管理办公室， 治疗AD的潜在治疗剂。

Project Summary/Abstract Alzheimer`s Disease (AD), is a severe neurodegenerative disorder characterized by progressive memory loss, language deficits, personality changes, and dementia, most prevalent in patents over 65. Current treatments only have modest and temporary effects on cognitive decline, highlighting the need for novel treatments which can halt the progression of AD. While familiar AD is caused by autosomal dominant mutations in amyloid precursor protein and presenilin, the majority of AD cases are considered sporadic with a combination of both genetic and environmental risk factors. However, amyloid-beta (Aβ) plaques, hyper-phosphorylated tau neurofibrillary tangles (NTs), and severe gliosis are all neurodegenerative hallmarks common to all forms of AD. The receptor for advanced glycation end-products (RAGE) is upregulated in the brains of AD patients and interacts directly with Aβ in neuronal, immune and vascular cells. RAGE signaling can also induce hyper-phosphorylation of tau and is implicated in the pathogenesis of co-morbid risk factors for sporadic AD (e.g. cardiovascular disease and diabetes). Thus, RAGE is a prime target for therapeutic development in both familial and sporadic AD. The RAGE pre-mRNA yields mutually exclusive alternative splice variants that are either, membrane- bound and capable of signaling (mbRAGE), or truncated and endogenously secreted to clear RAGE ligands without signaling (esRAGE). In AD mice, overexpression of mbRAGE accelerated the pathologic, cognitive, and behavioral hallmarks of AD, and infusion of synthetic esRAGE decreased Aβ and NT pathology as well as improved learning/memory and synaptic function. Thus, a novel strategy to reduce RAGE signaling is develop compounds called splice modulating oligonucleotides (SMOs) capable of controlling pre-mRNA alternative splicing to decrease expression of mbRAGE and simultaneously increase expression of esRAGE. SMOs are a class of synthetic RNA based compounds that sterically block or weaken interactions between elements of the splicing machinery and the pre-mRNA. SMOs bind to their targets with exceptional potency, specificity, long duration of action and negligible off-target effects. Previous pharmacological approaches to target ligand- RAGE interactions have been hampered by off-target effects or a short half-life, and none have the dual mechanism of both increasing natural esRAGE and decreasing mbRAGE isoform expression proposed here. SMOs targeting RAGE will be designed in silico and refined for potency and specificity, first in vitro cell culture (Aim 1) to select lead SMO drug candidates. Each SMO candidate will then be characterized and further optimized for safety and potency in vivo in RAGE transgenic mice (Aim 2). Finally, SMOs will be evaluated as therapeutics in an acute streptozotocin (STZ) induced model of AD for prevention/reversal of cognitive deficits and tau hyper-phosphorylation (Aim 3). The ultimate goal is to develop a lead RAGE SMO as potential therapeutic for the treatment of AD.