Activation of Neuronal Degradative Pathways to Ameliorate Prion Disease

激活神经元降解途径以改善朊病毒病

• 批准号: 10855708
• 负责人: SANDRA E Encalada
• 金额: $ 87.2万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10855708
• 关键词: Agreement; Alzheimer' s Disease; Alzheimer' s disease patient; Alzheimer' s disease related dementia; Automobile Driving; Autophagocytosis; Axon; Axonal Transport; Behavioral; Biological; Brain; CRISPR-mediated transcriptional activation; Candidate Disease Gene; Categories; Cell model; Cells; Clinical; Creutzfeldt-Jakob Syndrome; Cytoskeleton; Data; Degradation Pathway; Deposition; Disease; Drug Targeting; Elements; Endosomes; Excision; Fatal Familial Insomnia; Functional disorder; Gene Targeting; Genes; Genetic; Gerstmann-Straussler-Scheinker Disease; Gliosis; Goals; Grant; Human; Impaired cognition; Impairment; Investigation; Kinesin; Lesion; Lipids; Lysosomes; Modeling; Molecular Motors; Movement; Mus; Mutation; Neurodegenerative Disorders; Neurologic; Neuronal Dysfunction; Neurons; Organelles; Other Genetics; Pathogenicity; Pathologic; Pathology; Pathway interactions; Penetrance; Pharmaceutical Preparations; Poisoning; PrP; PrP gene; Preclinical Testing; Prion Diseases; Prions; Property; Rare Diseases; Sleep disturbances; Sorting; Structure; Swelling; Testing; Therapeutic; Toxic effect; Up-Regulation; Vesicle; Work; aggregation pathway; clinical candidate; disease phenotype; efficacy evaluation; efficacy testing; functional restoration; genome wide screen; improved; improved outcome; insight; misfolded protein; mouse model; mutant; nanomolar; neuron loss; neurotoxic; pharmacologic; prevent; protein aggregation; screening; small molecule; therapy development; trafficking; vesicle transport; virtual

ABSTRACT Prionopathies are rare human neurodegenerative diseases characterized by spongiform change, gliosis, and by the deposition of misfolded prion protein (PrP) aggregates inside and outside of neurons across the brain. While cellular mechanisms remain largely undefined, evidence points toward a particular vulnerability of axons to the formation of misfolded protein aggregates, and their accumulation inside lysosome-like compartments suggests defective lysosomal degradative pathways in axons. Indeed, axonal dystrophies with enlarged endosomes occur early in disease in virtually all neurodegenerative diseases including Alzheimer’s Disease and Alzheimer’s Disease related dementias, where lysosomal dysfunction is well-recognized. Compelling evidence shows that PrP aggregates impair neuronal function by driving the accumulation of organelles/vesicles and cytoskeletal elements, thus poisoning axonal transport. This application builds on our previous findings that identified an endolysosomal pathway unique to axons, that promotes the initial stages of formation of misfolded mutant PrP aggregates inside enlarged endolysosome structures that do not acidify and thus fail to degrade misfolded and aggregated mutant PrP from axons, indicating impaired lysosomal degradation. We showed that in this axonal rapid endosomal sorting and transport-dependent aggregation (ARESTA) pathway, the molecular motor kinesin- 1C (KIF5C) transports vesicles carrying pathogenic and misfolded mutant PrP into axons resulting in neurotoxic axonal dystrophies filled with PrP aggregates inside endosomes that we term ‘endoggresomes’. Reducing the function of ARESTA genes, including kinesin-1, efficiently inhibits mutant PrP endoggresome formation and restores neuronal function. Furthermore, we have identified and tested a lysosomal flux activator (LFA) small molecule that efficiently inhibits and/or clears mutant PrP aggregate-containing endoggresomes from axons, restoring neuronal function. These findings form the premise of the central hypothesis of this grant that states that targeting neurotoxic axonal aggregates by genetic inhibition of ARESTA or by pharmacologic activation of lysosomal flux, prevents the formation of misfolded PrP aggregates and/or clears them, and ameliorates disease phenotypes in cellular and mouse models of prion disease. Our main LFA candidate molecule degrades PrP aggregates in the lower nanomolar range, does not show any overt signs of toxicity in mice, and has brain penetrance. The proposed aims will test the efficacy of LFAs in cellular (neuronal) and mouse models of familial and infectious prion disease. We will also identify the mechanisms of action (MoA) of LFAs. Our findings reveal a therapeutic strategy to treat prionopathies by genetic and pharmacological activation of macroautophagy. As lysosomal clearance is a common pathway impaired in Alzheimer’s Disease and Alzheimer’s Disease related dementias, our findings are expected to also be relevant to the treatment of these disorders.

摘要 原始性疾病是一种罕见的人类神经退行性疾病，其特征是海绵状改变、胶质细胞增多症和 错误折叠的Prion蛋白(PrP)在整个大脑的神经元内外聚集。而当 细胞机制在很大程度上仍然不清楚，有证据表明轴突对 错误折叠的蛋白质聚集体的形成及其在溶酶体样隔间内的积聚表明 轴突中有缺陷的溶酶体降解途径。确实，轴突营养不良伴内小体增大。 几乎所有神经退行性疾病的早期发病，包括阿尔茨海默病和阿尔茨海默病 疾病相关性痴呆，其中溶酶体功能障碍是公认的。令人信服的证据表明 PRP聚集体通过驱动细胞器/囊泡和细胞骨架的积累而损害神经元功能 元素，从而毒化轴突运输。此应用程序构建在我们之前的发现基础上，该发现标识了 轴突特有的内溶酶体途径，促进错误折叠突变体PrP形成的初始阶段 扩大的内溶酶体结构内的聚集体，不会酸化，因此不能降解错误折叠和 突变体PrP从轴突聚集，表明溶酶体降解受损。我们在这个轴突中展示了 快速内体分选和转运依赖聚集(ARESTA)途径，分子马达激动素- 1C(KIF5C)将携带致病和错误折叠突变PrP的囊泡运送到轴突中，导致神经毒性 充满PrP的轴突营养不良在内小体内聚集，我们称之为内小体。减少了 ARESTA基因的功能，包括Kinesin-1，有效地抑制突变的PrP内生体的形成和 恢复神经功能。此外，我们还鉴定并测试了一种溶酶体通量激活剂(LFA)小分子 有效地抑制和/或清除突变型PrP聚合体内含轴突内糖体的分子， 恢复神经功能。这些发现构成了这项拨款的中心假设的前提，即 通过Aresta的遗传抑制或通过药物激活Aresta靶向神经毒性轴突聚集体 溶酶体通量，防止错误折叠的PrP聚集体的形成和/或清除它们，并改善疾病 Pron病细胞模型和小鼠模型的表型。我们主要的LFA候选分子降解PrP 聚集体在较低的纳摩尔范围内，在小鼠身上没有任何明显的毒性迹象，并且有大脑 洞察力。拟议的AIMS将在细胞(神经元)和小鼠家族性白血病模型中测试LFAs的有效性。 和传染性普恩病毒病。我们还将确定LFA的作用机制(MOA)。我们的发现揭示了 一种通过基因和药物激活巨噬细胞来治疗原发疾病的治疗策略。AS 溶酶体清除是阿尔茨海默病及相关阿尔茨海默病的常见途径 对于痴呆症，我们的发现预计也将与这些疾病的治疗相关。