REPURPOSING PRION PROTEINS AS TRANSLATIONAL AMYLOID-TARGETING THERAPEUTICS FOR ALZHEIMER'S DISEASE

重新利用朊病毒蛋白作为阿尔茨海默病的翻译淀粉样蛋白靶向疗法

基本信息

批准号：
9891697
负责人：
Zachary Alan Levine
金额：
$ 12.65万
依托单位：
YALE 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：
2020-06-15 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/9891697
关键词：
Affect Affinity Alanine Alzheimer&apos s Disease Alzheimer&apos s disease related dementia Alzheimer&apos s disease therapeutic Amino Acids Amyloid Amyloid Fibrils Amyloid beta-Protein Animal Model Binding Binding Proteins Binding Sites Biological Assay Biological Models Biomimetics Biophysics Blood - brain barrier anatomy Brain Cells Chemicals Chromatography Complex Dementia Dot Immunoblotting Educational workshop Enzyme-Linked Immunosorbent Assay Follow-Up Studies Goals Heterogeneity Human Immersion Immunoassay Impaired cognition Impairment In Vitro Joints Laboratories Length Ligand Binding Link Lipids Liquid substance Measures Mediating Membrane Proteins Mentors Methodology Methods Modeling Molecular Molecular Conformation Molecular Sieve Chromatography NMR Spectroscopy Nerve Degeneration Neurodegenerative Disorders Neurons Noise Optics Pathologic Pathway interactions Peptide aptamers Peptides Physiological PrP Prion Diseases Property Protein Binding Domain Proteins Protocols documentation Rattus Research Resources Sampling Scanning Senile Plaques Severities Severity of illness Side Signal Transduction Spectrum Analysis Speed Structural Models Structure Synapses System Tertiary Protein Structure Testing Thermodynamics Tissues Toxic effect Translations Water abeta oligomer abeta toxicity age related amyloid formation aptamer base biophysical analysis clinically relevant cost effective cytotoxicity design effectiveness evaluation experimental study genome wide screen in vivo meetings molecular dynamics monomer next generation protein complex relating to nervous system screening simulation synaptic function targeted treatment tau Proteins tau aggregation therapeutic target

项目摘要

PROJECT SUMMARY Cognitive decline in Alzheimer’s Disease (AD) and many other age-related dementias have long been associated with the presence of insoluble amyloid plaques that disrupt normal synaptic functioning. However, more recent studies have revealed that synapse impairment from AD is much more potently associated with soluble amyloid-beta oligomers (abo), rather than from insoluble fibrils. Soluble oligomers are structurally irregular and promiscuously bind to membrane proteins, thereby dysregulating downstream amyloid assemblies such as tau. These properties, unfortunately, have also made it difficult to experimentally characterize abo, since oligomeric states can be transient or otherwise observed as noise. Multiple genome-wide screening methods have identified cellular prion protein (PrPc) as a putative target of abo, and subsequent studies have confirmed a pathophysiological pathway in AD involving abo-PrPc binding. Interestingly, ab monomers and insoluble fibrils do not bind to PrPc, thus the binding domain of PrPc can be exploited in peptide aptamers to target and stabilize abo. Here, we seek to utilize these interactions by designing biomimetic PrPc peptides that complex soluble abo. Molecular simulations and amyloid-characterizing experiments will be combined to optimize aptamer-abo interactions in order to abrogate binding of abo to PrPc. While aptamers are unlikely to serve as an AD therapeutic, amyloid-targeting PrPc peptides can guide the construction of next-generation agents that cross the blood-brain barrier and potently inhibit the toxicity of abo. Similarly, identification of abo by aptamers can potentially enable the tracking of soluble oligomers through fluorescent tagging during the formation of insoluble fibrils. Given that there are few, if any methodologies to target abo, this proposal would seek to isolate soluble oligomers and identify the limitations of their interactions with membrane proteins. In order to synergistically combine molecular dynamics simulations with experiments, mentoring will be carried out on the use of NMR spectroscopy, chromatography, and ligand-binding assays to measure oligomer structure, size, and the ability to bind PrP proteins, respectively. Mentoring will include regular meetings, coursework, workshops, and immersion in the laboratory of the primary mentor. Additionally, this study will seek to bridge basic biophysical research with clinically-relevant systems through the translation of model aptamers from simulations into experiments. Results will be connected to and interpreted in the context of Alzheimer’s Disease. The protocols and products developed as a result of this study will subsequently inform follow-up studies of soluble amyloid toxicity in live cells, with extensions to multiple neurodegenerative diseases.

项目概要阿尔茨海默氏病（AD）和许多其他与年龄相关的痴呆症的认知能力下降长期以来一直被认为是与破坏正常突触功能的不溶性淀粉样斑块的存在有关。最近的研究表明，AD 引起的突触损伤更可能与以下因素相关：可溶性β-淀粉样蛋白寡聚体（abo），而不是来自不溶性原纤维，在结构上是可溶性寡聚体。不规则且混杂地与膜蛋白结合，从而失调下游淀粉样蛋白组装不幸的是，这些特性也使得通过实验表征 abo 变得困难，因为寡聚状态可以是瞬态的或以其他方式观察为噪声。多种全基因组筛选方法已将细胞朊病毒蛋白 (PrPc) 确定为假定目标 abo 的存在，随后的研究证实了 AD 中涉及 abo-PrPc 结合的病理生理学途径。提示，ab单体和不溶性原纤维不与PrPc结合，因此PrPc的结合域可以是在这里，我们寻求通过设计来利用这些相互作用。与可溶性 abo 复合的仿生 PrPc 肽。分子模拟和淀粉样蛋白表征。将结合实验来优化适体-abo 相互作用，以消除 abo 与 PrPc 的结合。虽然适体不太可能作为 AD 治疗药物，但靶向淀粉样蛋白的 PrPc 肽可以指导构建能够穿过血脑屏障并有效抑制 abo 毒性的下一代药物。类似地，通过适体识别 abo 可能能够通过以下方式追踪可溶性寡聚体：鉴于在不溶性原纤维的形成过程中进行荧光标记，即使有的话，也很少有方法。以 abo 为目标，该提案将寻求分离可溶性低聚物并确定其相互作用的局限性与膜蛋白。为了将分子动力学模拟与实验相结合，指导将是使用核磁共振波谱、色谱和配体结合测定来测量低聚物指导将分别包括结构、大小和结合 PrP 蛋白的能力。此外，本研究还将寻求主要导师的课程作业、研讨会和实验室沉浸。通过模型适体的翻译将基础生物物理研究与临床相关系统联系起来从模拟到实验的结果将与阿尔茨海默氏症联系起来并在其中进行解释。这项研究的结果开发的方案和产品将随后为后续行动提供信息。研究活细胞中可溶性淀粉样蛋白毒性，并扩展到多种神经退行性疾病。