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 全球有数百万人，是美国医疗保健的主要负担。广告是世界上最常见的事业 痴呆症，是美国第六大致死原因。帕金森病是一种进展到 认知缺陷。目前尚无有效的治疗AD或PD的方法；甚至一些用于治疗AD或PD的药物 改善症状，如帕金森病，会导致严重的长期副作用，可能比疾病更严重 它本身。在AD和PD中，淀粉样蛋白-β(A-β)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 认知或运动障碍。在本申请中，我们设想了一种创新的遗传工具(名为Autophagon，或 AFN)将被开发为一种基因疗法，有助于针对有毒的Aβ或α-Syn物种进行自噬，以及 在阿尔茨海默病和帕金森病中通常受损的重要降解途径。AFN将以合成基因为特色 帮助隔离Aβ或α-Syn聚集体的片段，并将它们运送到自噬小泡进行清除 来自神经元的。利用病毒载体，我们将通过特定的立体定位将AFN传递到细胞或小鼠的大脑 打针。首先，我们将使用2D和3D(脑器官)神经元培养物在体外测试AFN。 携带APP/PSEN1或α-SYN突变的IPSC分别驱动Aβ或α-SYN聚集，并评估 AFN抑制聚集形成和神经元毒性的能力。其次，我们将使用 AD(具有导致Aβ聚集的人类APP突变的基因敲入)和PD(表达 容易聚集的突变体人α-Syn)来测试黄曲霉毒素的治疗潜力及其预防能力 体内的聚集体形成、神经变性和认知或运动障碍。如果成功，建议的 该项目将通过开发有效的潜力，对神经退行性疾病领域产生重大影响 AD和PD的基因治疗，并为下一步的临床前和临床测试奠定了基础 建议的治疗方法。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退行性疾病。

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.