Novel Cytoskeletal Stabilizers as Potential Treatments for Limbic Lewy Body Disorders

新型细胞骨架稳定剂作为边缘系统路易体疾病的潜在治疗方法

• 批准号: 10040472
• 负责人: ALEEM GANGJEE
• 金额: $ 37.95万
• 依托单位: DUQUESNE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10040472
• 关键词: Affect; Age-Months; Alzheimer' s disease patient; Animal Model; Anterior; Antibodies; Anxiety; Architecture; Autopsy; Axonal Transport; Behavioral; Binding; Biological Assay; Brain; Brain region; Cell Count; Cell Death; Cell model; Cells; Clinic; Clinical; Clinical Treatment; Cognition; Cognitive; Colchicine; Complex; Confocal Microscopy; Cytoskeleton; Data; Deterioration; Diet; Diffuse; Disease Progression; Docking; Dose; Drug Kinetics; Dyes; Embryo; Emotions; Epothilones; Equilibrium; Exposure to; Family; Female; Funding; Future; Goals; Hand; High Pressure Liquid Chromatography; Hippocampus (Brain); Histologic; Human; Immunoblotting; In Vitro; Infusion procedures; Intervention; Intervention Studies; Lead; Letters; Lewy Body Dementia; Lewy Body Disease; Lewy body pathology; Light; Limbic System; Measures; Memory; Metabolic; Microtubule Depolymerization; Microtubule Stabilization; Microtubule stabilizing agent; Microtubules; Modality; Modeling; Movement; Mus; Nerve Degeneration; Neurodegenerative Disorders; Neuroglia; Neurons; Operative Surgical Procedures; Paclitaxel; Parkinson Disease; Pathologic; Pathology; Patients; Pharmacology; Plant Roots; Procedures; Property; Proteins; Recovery; Reporting; Research; Research Design; Role; Safety; Smell Perception; Stains; Structure; Syndrome; Testing; Toxic effect; Translations; Tubulin; Ubiquitin; Vinca; Vincristine; alpha Tubulin; alpha synuclein; analog; base; behavior test; blood-brain barrier penetration; complement C2a; cost; density; design; disorder risk; drug testing; experimental study; gender difference; human disease; human mortality; human tissue; improved; in silico; in vivo; male; melting; men; molecular modeling; monomer; motor behavior; motor deficit; mouse model; novel; olfactory bulb; olfactory nuclei; polymerization; postnatal; pre-clinical; protein aggregation; proteotoxicity; synucleinopathy; tool

Abstract Lewy body disorders are a family of fatal neurodegenerative conditions with α-synucleinopathic inclusions spreading across brain regions involved in smell, affect, movement, and cognition. There are ~1.3M patients living in the US with dementia with Lewy bodies (DLB) and another ~4M patients living with Parkinson's disease (PD). In addition, an estimated ~50% of Alzheimer's disease patients display coexisting Lewy pathology (ADLB). If therapies could be developed that slow the onset and spread of α-synucleinopathy, neurodegeneration might be mitigated and deterioration of function would be delayed in millions of patients. Recent studies reveal that α-synuclein is a novel microtubule (MT) dynamase, disrupting axonal transport across microtubules (MTs) in its aggregated form. Thus, tipping MT dynamicity towards stabilization has emerged as a potential new treatment for neurodegenerative disorders. Accordingly, we observed significant loss of detyrosinated (stable) α-tubulin in the olfactory bulb of men with Lewy body disorders. The olfactory bulb/anterior olfactory nucleus (OB/AON) complex is closely connected with the limbic system and is one of the earliest regions to display α-synucleinopathic inclusions in this family of conditions. We have also recently discovered that two novel MT stabilizers (AG161-41 & 47) reduce the emergence of α-synuclein aggregates (up to a remarkable 48%) in primary hippocampal cultures exposed to preformed α-synuclein fibrils. To our knowledge, this observation is unique, as other MT stabilizers have not been demonstrated to mitigate Lewy pathology. Our human tissue and primary culture pilot data make a compelling argument for further proof-of-concept studies. Thus, we will synthesize improved analogs and test the hypothesis that MT stabilization tempers the pathological sequelae of exposure to α-synuclein fibrils in cellular and animal models of limbic Lewy pathology. In Aim 1, we will synthesize additional AG161-41 & 47 and 12 novel analogs of AG161-47. In Aims 2A-C, we will determine if tipping the balance towards MT stabilization with these 14 compounds reduces inclusion density, cell loss, protein aggregation, and markers of MT destabilization in primary limbic neuron cultures treated with α-synuclein fibrils. In Aim 2D, we will evaluate the compounds in assays of tubulin assembly and displacement with vincristine. In Aim 3A, we will characterize the pharmacokinetic properties of AG161- 47 and 4 of the most promising analogs from Aim 2. In Aim 3B, we will test the neuroprotective potential of 5 doses of the single most effective, safe analog from Aims 2-3A, in mice infused with α-synuclein fibrils in the OB/AON. Cognitive, anxiety, olfactory, and motor behavior will be assessed at baseline and monthly intervals. α-synuclein inclusions and cell counts will be measured with unbiased stereological tools. Aim 3B will identify a pharmacologically active dose (PAD) that mitigates inclusions, cell loss, protein aggregation, and MT destabilization. In Aim 3C, we will use this PAD in a longer-term, interventional study designed to test if the most promising candidate can rescue neurons after behavioral deficits emerge (e.g., smell loss emerges within 3 months). The assays of Aim 2B will be repeated. These proof-of-concept studies are designed to 1) pave the way for future SAR studies, 2) generate new hypotheses about the role of cytoskeletal stabilization in Lewy body disorders in vivo, and 3) help accelerate the translation of MT-modifying agents to the clinic.

摘要 路易体疾病是一个家族的致命性神经退行性疾病与α-突触核蛋白病包涵体扩散 涉及嗅觉、情感、运动和认知的大脑区域。美国约有130万患者患有 路易体痴呆（DLB）和另外约400万帕金森病（PD）患者。此外，一个 估计约50%的阿尔茨海默病患者表现出共存的路易病（ADLB）。如果治疗可以 开发减缓α-突触核蛋白病的发病和扩散，神经变性可能会减轻和恶化 数百万患者的功能将被延迟。最近的研究表明α-突触核蛋白是一种新的微管 动力酶，以其聚集的形式破坏穿过微管（MT）的轴突运输。因此， 已经成为神经变性疾病的潜在新治疗方法。因此我们 观察到路易体疾病患者嗅球中脱酪氨酸（稳定）α-微管蛋白的显著丢失。的 嗅球/前嗅核（OB/AON）复合体与边缘系统关系密切，是一种重要的 最早的区域显示α-突触核蛋白病包涵体在这个家庭的条件。我们最近还发现 两种新型MT稳定剂（AG 161 -41和47）减少了α-突触核蛋白聚集体的出现（高达48%） 在暴露于预先形成的α-突触核蛋白纤维的原代海马培养物中。据我们所知，这种观察是独一无二的， 因为其他MT稳定剂还没有被证明能减轻路易病。我们的人体组织和原代培养 试点数据为进一步的概念验证研究提供了令人信服的理由。因此，我们将合成改进的类似物， 并检验MT稳定化可以缓和暴露于α-突触核蛋白纤维的病理后遗症的假设， 边缘路易病理学的细胞和动物模型。在目标1中，我们将合成另外的AG 161 -41和47和12 AG 161 -47的新类似物。在目标2A-C中，我们将确定是否将平衡转向MT稳定， 14种化合物减少了原发性肝癌中的包涵体密度、细胞损失、蛋白质聚集和MT不稳定的标志物。 用α-突触核蛋白原纤维处理的边缘神经元培养物。在Aim 2D中，我们将在微管蛋白测定中评估化合物 用长春新碱组装和置换。在目标3A中，我们将表征AG 161 - 1000的药代动力学性质。 47和4个最有前途的类似物从目标2。在目标3B中，我们将测试5种剂量的 在OB/AON中输注α-突触核蛋白原纤维的小鼠中，来自目标2-3A的单一最有效、最安全的类似物。认知、 焦虑、嗅觉和运动行为将在基线和每月间隔进行评估。α-突触核蛋白包涵体和细胞 计数将用无偏体视学工具测量。目标3B将确定一个有效剂量（PAD） 其减轻内含物、细胞损失、蛋白质聚集和MT不稳定。在Aim 3C中，我们将在 一项长期的干预性研究，旨在测试最有前途的候选人是否可以在行为后拯救神经元。 出现缺陷（例如，3个月内出现嗅觉丧失）。将重复目标2B的测定。这些概念验证 研究的目的是：1）为未来的SAR研究铺平道路，2）产生关于细胞骨架作用的新假设， 稳定体内路易体疾病，和3）有助于加速MT修饰剂向临床的转化。