Enabling Efficient, Fast, Biocompatible Exosome Separation via Acoustofluidics

通过声流控技术实现高效、快速、生物相容性的外泌体分离

基本信息

批准号：
10456734
负责人：
Tony Jun Huang
金额：
$ 42.96万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10456734
关键词：
Acoustics Address Albumins Alzheimer&apos s Disease Antigen Presentation Benchmarking Biological Biological Markers Biological Process Biomedical Research Blood Blood Circulation Cell Culture Techniques Cell physiology Cells Centrifugation Cerebrospinal Fluid Chylomicrons Clinical Communities Containment of Biohazards Contracts DNA Devices Diagnosis Diagnostic Dimensions Disease Drug Delivery Systems Freezing Glass Health High Density Lipoproteins Human Resources Hybrids IDL lipoproteins Immunoglobulins In Vitro Industry Standard Kidney Diseases Lipoproteins Liquid substance Liver diseases Low-Density Lipoproteins Malignant Neoplasms Messenger RNA Methods MicroRNAs Microfluidics Molecular Molecular Profiling Monitor Morphology Neurodegenerative Disorders Parkinson Disease Patients Performance Phase Placenta Diseases Procedures Prognosis Property Proteins Recovery Research Research Personnel Saliva Sampling Series Serum Proteins Side Sorting - Cell Movement Speed Techniques Technology Therapeutic Time Training Transducers Urine Very low density lipoprotein Vesicle base biological research biomarker discovery biomaterial compatibility cell type circulating biomarkers clinical application cost density design disease diagnostic exosome extracellular vesicles improved innovation interactive feedback intercellular communication microfluidic technology nanoscale nanosized non-Native operation particle physical property point of care preservation prognostic tool

项目摘要

Abstract Exosomes are nanosized extracellular vesicles that contain biomolecules (DNA, mRNA, miRNA, and other functional proteins) from their cell of origin. Exosomes are secreted from nearly all cell types, and as a result, they are found in most biological fluids, including blood, saliva, urine, and cerebrospinal fluid. Over the past decade, the transfer of exosomal biomolecules to recipient cells has been implicated in a variety of biological processes. Consequently, exosomes have increasingly been the focus of many studies in biomedical research. Due to their distinct molecular signatures, exosomes have been identified as a potentially transformative circulating biomarker for the diagnosis and prognosis of multiple diseases, including cancer, neurodegenerative diseases (i.e., Parkinson’s and Alzheimer’s), as well as diseases of the kidney, liver, and placenta. In addition to diagnostic applications, exosomes are an ideal drug delivery system in many therapeutic applications. While the versatility of exosomes renders them an excellent candidate for a variety of biomedical applications, difficulties in the consistent, effective isolation of exosomes have greatly limited their utility. Current approaches for exosome isolation involve lengthy procedures, require highly trained personnel, suffer from low repeatability, low yield, low purity, and/or low post-sorting exosome integrity. As a result, there exists a critical need in the research communities for a simple, rapid, efficient, and biocompatible approach for isolating exosomes form biological fluids or in vitro cell culture. In this R01 project, we will address this unmet need by developing an acoustofluidic (i.e., the fusion of acoustics and microfluidics) platform for high-purity, high-yield, high-biocompatibility, automated exosome isolation. The proposed acoustofluidic technology will have the following features: 1) Automated exosome processing which reduces operator-to-operator variability and enables simple, consistent isolation results with improved biohazard containment; 2) Reduces the amount of time necessary to go from biofluid (e.g., 1 mL undiluted blood) to isolated exosomes (<5 min processing time vs ~8 hrs processing time with alternative technologies); 3) Higher exosome recovery rate (>90%) in comparison to benchmark technologies (5‒25%); 4) Greater exosome purity (>80%) in comparison to benchmark technologies (~33%); 5) Less contamination from other circulating factors, including non-native serum proteins (e.g., albumin and immunoglobulin) and particles with similar sizes, including various types of lipoproteins; 6) Low-cost and point- of-care design; and 7) ability to handle both large and small sample volumes (maximum sample volume: ~30 mL; minimum sample volume: ~10 µL), which is extremely challenging with existing approaches. With these unique features, the proposed acoustofluidic technology has the potential to greatly simplify and expedite workflows in exosome-related biomedical research and aid in the discovery of new exosomal biomarkers.

抽象的外泌体是含有生物分子的纳米细胞外蔬菜（DNA，mRNA，miRNA和其他功能蛋白）来自其原始细胞。外泌体从几乎所有细胞类型中分泌，结果它们是在大多数生物学液中发现的，包括血液，唾液，尿液和脑脊液。过去十年，在多种生物学中隐含了外泌体生物分子向受体细胞的转移过程。因此，外泌体已成为生物医学研究中许多研究的重点。由于其独特的分子特征，外泌体已被鉴定为潜在的变换循环生物标志物用于多种疾病的诊断和预后，包括癌症，神经退行性疾病（即帕金森氏症和阿尔茨海默氏症），以及肾脏，肝脏和斑点的疾病。此外诊断应用，外泌体是许多治疗应用中理想的药物输送系统。而外泌体的多功能性使它们成为各种生物医学应用的绝佳候选者在一致，有效的隔离中，外泌体极大地限制了它们的效用。当前的方法外泌体隔离涉及冗长的手术，需要高度训练的人员，重复性低，低点产量，低纯度和/或低术后外泌体完整性。结果，研究中存在着迫切的需求社区的一种简单，快速，高效和生物相容性的方法，用于隔离外泌体形成生物学液体或体外细胞培养。在这个R01项目中，我们将通过开发声流式来满足这种未满足的需求（即，声学和微流体的融合）高纯度，高产，高生物相容性的平台，自动外泌体隔离。拟议的声流动技术将具有以下功能：1）自动化外泌体处理，可降低操作员到操作员的可变性，并启用简单，一致的隔离结果，改善了生物危害遏制； 2）减少从生物流体（例如，1 ml未稀释的血液）到孤立的外泌体（<5分钟的加工时间vs 〜8小时处理时间使用替代技术）； 3）与基准相比，较高的外泌体回收率（> 90％）技术（5％至25％）； 4）与基准技术相比，外泌体纯度更大（> 80％）（〜33％）； 5）来自其他循环因子的污染物较少，包括非母血清蛋白（例如，白蛋白和白蛋白和免疫球蛋白）和具有相似大小的颗粒，包括各种类型的脂蛋白； 6）低成本和点 - 护理设计； 7）能够处理大型和小样品量的能力（最大样品体积：〜30 ML;最小样品量：〜10 µL），现有方法极为挑战。与这些独特的功能，拟议的声流动技术有可能极大地简化和加快与外泌体相关的生物医学研究中的工作流程，并有助于发现新的外泌体生物标志物。

项目成果

期刊论文数量（11）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Acoustofluidic methods in cell analysis.

DOI：
10.1016/j.trac.2019.06.034
发表时间：
2019-07
期刊：
Trends in analytical chemistry : TRAC
影响因子：
0
作者：
Yuliang Xie;Hunter Bachman;T. Huang
通讯作者：
Yuliang Xie;Hunter Bachman;T. Huang

Contactless, programmable acoustofluidic manipulation of objects on water.

DOI：
10.1039/c9lc00465c
发表时间：
2019-10
期刊：
Lab on a chip
影响因子：
6.1
作者：
Peiran Zhang;Chuyi Chen;Feng Guo;Julien Philippe;Yuyang Gu;Zhenhua Tian;Hunter Bachman;Liqiang Ren;Shujie Yang;Zhanwei Zhong;Po-Hsun Huang;N. Katsanis;K. Chakrabarty;T. Huang
通讯作者：
Peiran Zhang;Chuyi Chen;Feng Guo;Julien Philippe;Yuyang Gu;Zhenhua Tian;Hunter Bachman;Liqiang Ren;Shujie Yang;Zhanwei Zhong;Po-Hsun Huang;N. Katsanis;K. Chakrabarty;T. Huang

Harmonic acoustics for dynamic and selective particle manipulation.