A nanosized magnetic particle system for fast and efficient neuronal extracellular vesicle enrichment from plasma

概要/摘要 细胞外囊泡（EV）是纳米囊泡，其含有表面标记物，并且具有细胞和细胞状态特异性。 货物的组织（和细胞）的起源，反映了组织或细胞的生理状态。最重要的是， 神经元EV可以通过血脑屏障（BBB）进入血液循环， 直接接触大脑的致病过程新兴的临床研究表明，神经元EV 在血液中携带病原生物标记物，可以检测和预测神经系统疾病，如阿尔茨海默氏症 疾病、帕金森病、肌萎缩侧索硬化症和创伤性脑损伤等。因此， 从外周血中富集神经元EV提供了一种微创的方式来采样 神经系统疾病的脑组织成分。为了释放神经元电动汽车的巨大潜力， 神经系统疾病诊断应用，从血浆中快速有效富集神经元EV 是一个先决条件。然而，从血浆中有效分离神经元EV存在重大挑战， 并且没有用于从血液中分离神经元EV的市售技术/试剂盒 等离子体 在此，我们建议开发一种神经元EVs纳米探针（neuron-NPs）技术，用于高产量分离神经元EVs。 直接从血浆样品中提取神经元EV作为研究专用试剂盒。我们将开发和优化这些神经元- 用于在缓冲系统中快速有效地磁性分离神经元EV的NP（目的1）。Aim 2将在 血浆样品中神经元-NP技术的分析验证。成功实现这些目标将产生 一个先进的专有技术平台，用于各种基础和临床研究中的神经元EV分离 神经系统疾病第二阶段的活动将集中在扩大神经元电动汽车的制造 nanoprobes试剂盒和开发集成神经元的高通量自动样品处理系统， NPs试剂盒。

SUMMARY/Abstract Extracellular vesicles (EVs) are nanovesicles, which contain surface markers, and cell- and cell-state-specific cargo of their tissue (and cell) of origin, reflecting the tissue or cell’s physiological state. Most importantly, neuronal EVs can pass the blood-brain barrier (BBB) into blood circulation and provide a new approach of gaining direct access to brain pathogenic processes. Emerging clinical studies show that neuronal EVs in blood carry pathogenic biomarkers that can detect and predict neurological diseases such as, Alzheimer’s disease, Parkinson disease, amyotrophic lateral sclerosis, and traumatic brain injury, etc. Therefore, enrichment of neuronal EVs from peripheral blood provides a minimally invasive way to sample components of brain tissue for neurological diseases. To unleash neuronal EVs’ great potential in neurological diseases diagnosis applications, rapid and efficient enrichment of neuronal EVs from blood plasma is a prerequisite. However, there is major challenge to efficiently isolate neuronal EVs from blood plasma, and there are no commercially available technologies/kits for the isolation of neuronal EVs from blood plasma. Herein, we propose to develop a neuronal EVs nanoprobes (neuron-NPs) technology for high-yield isolation of neuronal EV directly from plasma sample as a research use only kit. We will develop and optimize these neuron- NPs for rapid and efficient magnetic isolation of neuronal EVs in a buffer system (Aim 1). The Aim 2 will perform analytical validation of neuron-NP technology in plasma samples. Successful completion of these Aims will yield an advanced proprietary technology platform for neuronal EVs isolations in basic and clinical research for various neurological diseases. Phase II activities will be focused on scaling up the manufacturing of neuronal EVs nanoprobes kit and developing high-throughput automatic sample processing system integrated with neuron- NPs kit.