Targeted antibody-conjugated magnetic nanoparticles for the treatment of Alzheimer's disease

摘要/项目摘要 阿尔茨海默病(AD)是痴呆症最常见的形式，其特征是进行性记忆丧失和 影响着500多万美国人的认知障碍。AD被假设是由于积累了 大脑中有两种致病蛋白质。一种是细胞外淀粉样蛋白β(Aβ)的积累，另一种是细胞外淀粉样蛋白的积累 是过度磷酸化的tau蛋白在细胞内积累，最终导致聚集，称为 神经原纤维缠结。淀粉样蛋白的聚集阻止了正常的细胞信号通路，而神经原纤维 抑制向神经元输送营养物质，这两者最终都会导致神经元死亡。以前的多个AD 临床试验使用治疗性抗Aβ抗体靶向致病的Aβ物种。然而，以前的失败 临床试验表明，我们目前对AD和AN的发病机制的了解存在差距 迫切需要开发新的安全治疗方法，这种方法可在 疾病的早期阶段。在以前的临床试验中，一个主要的副作用是长期存在高效价 大脑中的抗A-β抗体会触发炎症反应和其他不良副作用，即淀粉样蛋白- 相关影像异常包括微小出血(ARIA-H)和水肿(ARIA-E)。因为最近的结果 从阿杜卡努单抗临床试验显示出巨大的前景，迫切需要发展 一种减少这些副作用的技术。为了应对这一挑战，这可能会导致 在之前的药物试验中，我们开发了超顺磁性氧化铁纳米颗粒，与 与Aβ多肽结合并聚集Aβ物种的抗Aβ抗体。这些粒子是顺磁性的，它 允许它们在体外通过外部磁场被移除。验证抗A-β的有效性和安全性 抗体偶联SPION，我们将使用AD的3D人类神经细胞培养模型，这是我们的实验室 先前开发的和转基因AD小鼠模型。结合这两种技术，我们设计了一种 外磁引导清除抗A-β抗体快速清除A-β的方法学 结合型SPION在AD三维细胞培养中的应用3D细胞培养模型将主要用于测试疗效 抗Aβ抗体对β驱动的tau病理的影响，而转基因AD小鼠将用于 在体内评估其疗效和潜在毒性。AIM 1评估带有SPION的静电磁铁的使用 在AD的3D培养模型和AD小鼠模型中减少Aβ的种类。目的是进一步调查 3D培养模型中Aβ去除的下游tau效应。目标2将研究使用 交变磁场通过血脑屏障运送与SPION结合的治疗性抗体 在AD 5XFAD小鼠模型中。第二个目标对这项技术的可行性产生了巨大的影响，因为 一种新的治疗途径，不仅治疗AD，而且为各种神经系统疾病提供大分子药物 疾病。最终，这项工作的结果将导致AD AS新疗法的潜在开发 为当前AD药物和疗法的制定和应用提供了一种新的方法。

ABSTRACT/PROJECT SUMMARY Alzheimer’s disease (AD) is the most common form of dementia, characterized by progressive memory loss and cognitive disturbances affecting over 5 million Americans. AD is hypothesized to be due to the accumulation of two pathogenic proteins in the brain. One is the accumulation of amyloid-β (Aβ) outside of cells and the second is the accumulation of hyperphosphorylated tau protein inside cells that eventually leads to aggregates called neurofibrillary tangles. The aggregation of amyloid prevents normal cell signaling pathways while neurofibrillary inhibit nutrient delivery to the neurons, both of which ultimately leads to neuronal death. Multiple AD previous clinical trials target pathogenic Aβ species using therapeutic anti-Aβ antibodies. However, previous failures in clinical trials demonstrate a gap in knowledge in our current understanding of the pathogenesis of AD and an immediate need for the development of new safe therapeutic approaches, which can be applicable at the very early stage of the disease. One major side effect in previous clinical trials is the chronic presence of high-titer anti-Aβ antibodies in brains triggers inflammatory responses and other undesirable side effects, namely amyloid- related imaging abnormalities including microhemorrhages (ARIA-H) and edema (ARIA-E). Since recent results from the aducanumab clinical trial is showing immense promise, there is an urgent need for the development of a technology to reduce these side effects. To address this challenge, which may contribute to the failures of these previous current drug trials, we developed superparamagnetic iron oxide nanoparticles conjugated with anti-Aβ antibodies that bind to Aβ peptides and aggregated Aβ species. These particles are paramagnetic, which allows them to be removed by an external magnetic field in vitro. To validate the efficacy and safety of anti-Aβ antibody conjugated SPIONs, we will use both 3D human neural cell culture models of AD, which our lab developed previously, and transgenic AD mouse models. Combining these two technologies, we devised a methodology to rapidly remove Aβ species using external magnetic force guided removal of anti-Aβ antibody conjugated SPIONs in 3D cell culture of AD. The 3D cell culture model will be mostly used for testing efficacy and the impact of the anti-Aβ antibody on Aβ-driven tau pathology while transgenic AD mice will be used to assess the efficacy and potential toxicity in vivo. Aim 1 evaluates the use of a static magnet with SPIONs with to reduce Aβ species in 3D culture model of AD and in an AD mouse model. The aim further investigates the downstream tau effects of Aβ removal in the 3D culture model. Aim 2 will examine the potential of using an alternating magnetic field to deliver therapeutic antibodies conjugated to SPIONs across the blood-brain barrier in the AD 5XFAD mouse model. This second aim has tremendous impact on the feasibility of this technology as a new therapeutic avenue not only for AD, but to deliver large molecule drugs for a variety of neurological diseases. Ultimately, the results of this work will lead to the potential development of a new therapy for AD as well as a new method for formulating and applying current AD drugs and therapeutics.