Autophagon: an Autophagy-Functionalizing Gene Therapy Tool for Neurodegenerative Diseases

项目摘要 神经退行性疾病，特别是阿尔茨海默病（AD）和帕金森病（PD），影响 全球数百万人，是美国的主要医疗负担。AD是世界上最常见的病因 痴呆症，是美国第六大死亡原因。PD是进展至 认知缺陷目前还没有有效的治疗AD或PD的方法;甚至一些用于治疗AD或PD的药物也是如此。 改善症状，如在PD，导致严重的长期副作用，可能比疾病更糟 本身在AD和PD中，β淀粉样蛋白（Aβ）或α-突触核蛋白（α-Syn）的积累及其随后的聚集， 引起神经元毒性，包括神经炎症、突触缺陷和神经变性，导致 认知或运动缺陷。在这个应用中，我们设想了一个创新的遗传工具（名为Autophagon，或 AFN）将被开发为一种基因疗法，帮助靶向毒性Aβ或α-Syn物种进行自噬， 在AD和PD中通常受损的重要降解途径。AFN将采用合成基因 片段，以帮助隔离Aβ或α-Syn聚集体并将其递送至自噬囊泡进行清除 从神经元。利用病毒载体，我们将通过特定的立体定位将AFN递送到细胞或小鼠脑中， 注射剂首先，我们将在体外使用来自以下的2D和3D（脑类器官）神经元培养物测试AFN： 携带APP/PSEN 1或α-Syn突变的iPSC分别驱动Aβ或α-Syn聚集，并评估 AFN抑制聚集体形成和神经元毒性的能力。其次，我们将使用小鼠模型， AD（具有引起Aβ聚集的人类APP突变的基因敲入）和PD（表达 聚集倾向突变的人α-Syn），以测试AFN的治疗潜力及其预防 聚集体形成、神经变性和体内认知或运动缺陷。如果成功，建议 该项目将对神经退行性疾病领域产生重大影响，通过开发有效的潜力， AD和PD的基因治疗，并为下一步的临床前和临床试验奠定基础。 建议治疗。它还可以为其他疾病的治疗开发提供概念验证。 与蛋白质聚集相关的神经变性疾病。

Project Summary Neurodegenerative diseases, especially Alzheimer’s Disease (AD) and Parkinson’s Disease (PD), affect millions of people globally and represent a US major healthcare burden. AD is the world’s most common cause of dementia, and is the sixth leading cause of death in the US. PD is motor disabling condition that advances to cognitive deficits. Currently there exist no effective therapies for AD or PD; even some of the drugs used to ameliorate symptoms, such as in PD, cause serious long-term side effects that may be worse than the disease itself. In AD and PD, accumulation of Amyloid-β (Aβ) or α-Synuclein (α-Syn) and their subsequent aggregation, cause neuronal toxicity, including neuroinflammation, synaptic deficits and neurodegeneration, leading to cognitive or motor deficits. In this application, we conceive an innovative genetic tool (named Autophagon, or AFN) to be developed as a gene therapy that helps target toxic Aβ or α-Syn species to autophagy, an important degradative pathway that is usually impaired in AD and PD. AFN will feature a synthetic gene fragment to help sequester Aβ or α-Syn aggregates and deliver them to the autophagic vesicles for clearance from the neurons. Using viral vectors, we will deliver AFN to cells or to mouse brain via specific stereotaxic injections. First, we will test AFN in vitro using 2D and 3D (brain organoid) neuronal cultures derived from iPSCs harboring APP/PSEN1 or α-Syn mutations that drive Aβ or α-Syn aggregation, respectively, and assess the ability of AFN to suppress aggregate formation and neuronal toxicity. Second, we will use mouse models of AD (with a genetic knock-in of human APP mutations that cause Aβ aggregation) and PD (expressing aggregation-prone mutant human α-Syn) to test the therapeutic potential of AFN and its ability to prevent aggregate formation, neurodegeneration, and cognitive or motor deficits in vivo. If successful, the proposed project will have a major impact on the neurodegenerative diseases field, by developing an effective potential gene therapy for AD and PD, and set the foundation for the next steps of preclinical and clinical testing of this suggested therapy. It may also provide a proof-of-concept for therapy development for other neurodegenerative disorders associated with protein aggregation.