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

基本信息

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
项目类别：
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&apos s Disease Alzheimer&apos s disease patient Alzheimer&apos 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

Agreement Alzheimer&apos s Disease Alzheimer&apos s disease patient Alzheimer&apos 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

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项目摘要

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.

抽象的 prion病是罕见的人类神经退行性疾病，其特征是赞助，神经胶质变化，并由整个大脑神经元内外的错误折叠的prion蛋白（PRP）聚集物的沉积。尽管细胞机制在很大程度上保持不确定，证据表明轴突的特殊脆弱性形成错误折叠的蛋白质聚集体及其在溶酶体样室内的积累表明轴突中有缺陷的溶酶体降解途径。实际上，轴突营养不良症发生增加几乎所有神经退行性疾病的疾病早期，包括阿尔茨海默氏病和阿尔茨海默氏病与疾病相关的痴呆症，溶酶体功能障碍被广泛认可。令人信服的证据表明 PRP聚集物通过驱动细胞器/囊泡和细胞骨架的积累来损害神经元功能元素，从而中毒轴突运输。此应用程序建立在我们以前确定的发现的基础上轴突独有的内溶性途径，促进了错误折叠突变体PRP的初始阶段内部内部的聚集体增加的内溶性结构不会酸化，因此无法降解错误折叠，并且轴突的聚集突变体PRP，表明溶酶体降解受损。我们在这个轴突中表明快速内体分选和转运依赖性聚集（ARESTA）途径，分子运动动力蛋白 - 1C（KIF5C）将带有致病性和折叠突变体PRP的蔬菜转运到轴突中，导致神经毒性轴突性肌营养不良，充满了内体内的PRP聚集体，我们称其为“内虫”。减少 Aresta基因（包括驱动蛋白1）的功能有效地抑制了突变的PRP内染料形成和恢复神经元功能。此外，我们已经确定并测试了溶酶体通量激活剂（LFA）小有效抑制和/或清除轴突中含有突变体PRP聚集的内核的分子，恢复神经元功能。这些发现构成了这笔赠款中心假设的前提通过对Aresta的遗传抑制或药物学激活来靶向神经毒性轴突骨料溶酶体通量，防止形成错误的PRP聚集体和/或清除它们，并改善疾病病毒疾病的细胞和小鼠模型中的表型。我们的主要LFA候选分子降低PRP 较低的纳摩尔范围的聚集体，没有显示小鼠毒性的明显迹象，并且有大脑外观。提出的目标将测试LFA在细胞（神经元）和小鼠家族模型中的效率和感染性prion病。我们还将确定LFA的作用机理（MOA）。我们的发现揭示了通过遗传和药物激活大量噬菌体治疗临界病的治疗策略。作为溶酶体清除是阿尔茨海默氏病和阿尔茨海默氏病有关的一种常见途径痴呆症，我们的发现也有望与这些疾病的治疗有关。