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

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

抽象的 Lewy身体疾病是一个致命的神经退行性疾病的家族，具有α-突触性核酸内包含物的扩散在涉及气味，情感，运动和认知的大脑区域之间。美国有约130万患者 Lewy身体（DLB）和另外约400万名帕金森氏病（PD）的痴呆症。另外，一个估计约有50％的阿尔茨海默氏病患者表现出并存的路易病病理学（ADLB）。如果可以治疗开发了慢慢α-突触核疾病的发作和扩散，神经退行性可能会得到降低，并且相对数百万患者的功能将延迟。最近的研究表明，α-突触核蛋白是一种新型微管（MT）动力学，以汇总形式破坏横跨微管（MTS）的轴突运输。那，小费mt动态稳定已成为神经退行性疾病的一种潜在新治疗方法。观察到在嗅球液体疾病的嗅球中，明显丧失了驱散（稳定）α-微管蛋白。这嗅球/前嗅觉核（OB/AON）复合物与边缘系统紧密相连，是一种最早在该疾病系列中显示α-突触核酸内聚物的区域。我们最近也发现了那两个新型的MT稳定剂（AG161-41和47）降低了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-核蛋白原纤维的原发性海马培养物中。据我们所知，这种观察是独一无二的由于其他MT稳定器尚未被证明可以减轻路易病理学。我们的人体组织和原发性培养飞行员数据为进一步的概念验证研究提供了令人信服的论点。这，我们将合成改进的类似物并检验以下假设：MT稳定温度是暴露于α-突触核蛋白原纤维中的病理后遗症。边缘路易病病理学的细胞和动物模型。在AIM 1中，我们将合成其他AG161-41和47和12 AG161-47的新型类似物。在AIMS 2A-C中，我们将确定是否将平衡降低到MT稳定下 14化合物降低了纳入密度，细胞损失，蛋白质聚集和MT在原发性稳定的标记用α-突触核蛋白原纤维处理的边缘神经元培养物。在AIM 2D中，我们将评估小管测定法的化合物带有vincristine的组装和位移。在AIM 3A中，我们将表征AG161-的药代动力学特性 AIM 2中最有前途的类似物中的47和4。在AIM 3B中，我们将测试5剂的神经保护势 AIMS 2-3A的最有效，最安全的类似物在OB/AON中感染了α-突触核蛋白原纤维的小鼠中。认知的，动画，嗅觉和运动行为将以基线和每月的间隔进行评估。 α-突触核蛋白夹杂物和细胞计数将使用公正的立体工具来衡量。 AIM 3B将识别药物活性剂量（PAD）减轻夹杂物，细胞丢失，蛋白质聚集和MT不稳定。在AIM 3C中，我们将在A中使用此垫长期，介入的研究旨在测试行为后最有希望的候选人是否可以挽救神经元出现碎片环境（例如，3个月内出现气味丧失）。 AIM 2B的萨斯将重复。这些概念证明研究设计为1）为未来的SAR研究铺平道路，2）关于细胞骨架的作用的新假设在体内Lewy体型疾病中的稳定性，3）有助于加速MT修饰药物向诊所的翻译。