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：
10187480
负责人：
Zachary Alan Levine
金额：
$ 12.73万
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 2022-06-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10187480
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 efficacy 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

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 efficacy 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

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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.

项目摘要长期以来与破坏正常突触功能的不溶性淀粉样斑块的存在有关。然而，最近的研究表明，AD的突触受损与可溶性淀粉样蛋白β低聚物（ABO），而不是来自不溶性纤维。可溶性低聚物在结构上是不规则且滥交与膜蛋白结合，从而使下游淀粉样蛋白组件失调例如tau。不幸的是，这些属性也使其难以实验地表征ABO，因为寡聚状态可以是短暂的，也可以视为噪声。多种基因组筛查方法已将细胞prion蛋白（PRPC）鉴定为推定靶标 ABO的以及随后的研究证实了涉及ABO-PRPC结合的AD的病理生理途径。有趣的是，AB单体和不溶性原纤维不与PRPC结合，因此PRPC的结合结构域可以是在肽适体中利用靶向和稳定ABO。在这里，我们试图通过设计来利用这些互动复杂的固体ABO的仿生PRPC肽。分子模拟和淀粉样蛋白特征将结合实验以优化APATMER-ABO相互作用，以消除ABO与PRPC的结合。虽然适体不太可能用作AD疗法，但靶向淀粉样蛋白的PRPC肽可以指导跨越血脑屏障并可能抑制ABO毒性的下一代药物的结构。同样，适体对ABO的识别可以有可能通过在不溶性原纤维形成过程中荧光标记。鉴于很少，如果有任何方法 Target Abo，该建议将寻求隔离固体寡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结合在一起，心理将是在使用NMR光谱，色谱法和配体结合测定方面进行了测量低聚物结构，大小和结合PRP蛋白的能力。指导将包括定期会议，课程，讲习班和沉浸在主要导师的实验室中。此外，这项研究将寻求通过模型适体的翻译与临床上的系统桥接基本的生物物理研究从模拟到实验。结果将在阿尔茨海默氏症的背景下连接并解释疾病。这项研究的结果将随后告知随访活细胞中固体淀粉样蛋白毒性的研究，并扩展到多种神经退行性疾病。