Fingerprinting Cancer via Multiparameter Extracellular Vesicle Analysis

项目摘要 电子生物科学（EBS）提出，与杰出教授亨利白色在合作， 犹他州大学，开发基于纳米孔的细胞外囊泡（EV）表征系统，能够 高分辨率、单囊泡、基于溶液的表征、分选和分离。电动汽车是纳米级的， 在正常生理条件下几乎所有哺乳动物细胞产生的细胞源性囊泡， 随着各种疾病状态（例如癌症、神经疾病、精神疾病）的产生率增加， 疾病等）。EV可以在大多数生物液体中找到，并且含有蛋白质，核酸和脂质分子 这与亲本细胞的细胞质内容物明显不同，表明外泌体负载不是一个重要的因素。 扩散或不受管制的过程。此外，EV通过将这些信息传递给细胞来促进细胞间的通信。 内容物进入受体细胞以改变靶细胞的表型和功能。除此之外，一般 作为一种重要的生理作用，EV在疾病发病机制中具有重要意义，并且具有作为 治疗靶点和药物载体。因此，这激发了人们对新方法的极大兴趣， 不仅表征EV的物理性质（大小、ζ电位、表面标记、核酸货物， 和浓度），但对开发新技术的兴趣增加， 准确地分选/隔离EV，使得可以以亚群特异性方式对内容物/货物进行分析。 EV表征和分选/分离领域的进展将极大地有助于理解 EV在人类健康中的一般作用，并且还有益于旨在在分子诊断中利用EV的努力， 生物标志物鉴定和癌症的靶向治疗。电动汽车特征和分选系统，EBS 和亨利白色教授将在这个项目中专门开发单粒子 EV样品的尺寸、zeta电位、荧光标记表面密度的真实分布的评估 标记物和核酸货物，沿着分离/收集特异性EV亚群的能力， 定义的生物物理参数，都在一个平台上。拟议系统的可行性将是 通过构建原型平台并展示其表征，分类和隔离EV的能力来证明 样品所得到的系统将具有与流式细胞术类似的易用性、通量和价格点， 动态光散射（DLS）、纳米颗粒跟踪分析（NTA）或单程连续脉冲技术， 具有与显微镜技术相当的分辨率，例如扫描或隧道电子显微镜（SEM或SEM）。 TEM）或原子力显微镜（AFM），产生一种最先进的仪器，它将直接 推动和推进电动汽车诊断领域的发展。

Project Summary Electronic BioSciences (EBS) proposes, in collaboration with Distinguished Professor Henry White at the University of Utah, to develop a nanopore-based extracellular vesicle (EV) characterization system capable of high-resolution, single-vesicle, solution-based characterization, sorting, and isolation. EVs are nanometer-scale, cell-derived vesicles that are produced by nearly all mammalian cells under normal physiological conditions and with increased production rates for various disease states (e.g. cancers, neurological diseases, psychiatric disorders, etc.). EVs can be found in most biological fluids and contain proteins, nucleic acids, and lipid molecules that are distinctly dissimilar to the parent cell’s cytoplasmic contents, indicating that exosome loading is not a diffusive or unregulated process. Furthermore, EVs facilitate intercellular communication by transferring these contents into recipient cells to alter the target cells’ phenotype and function. Thus, in addition to their general physiological roles, EVs have important implications in disease pathogenesis, and significant potential as therapeutic targets and drug carriers. Consequently, this has spurred a great deal of interest in new methods to not only characterize the physical properties of EVs (size, zeta potential, surface markers, nucleic acid cargo, and concentration), but increased interest in developing new techniques that have the ability to efficiently and accurately sort/isolate EVs such that the contents/cargo can be profiled in a subpopulation-specific manner. Advances in the areas of EV characterization and sorting/isolation would greatly assist the efforts to understand the general role of EVs in human health, and also benefit efforts aimed at utilizing EVs in molecular diagnostics, biomarker identification, and targeted therapies for cancer. The EV characterization and sorting system that EBS and Professor Henry White will be developing during this program will specifically enable single-particle assessments of an EV sample’s true distribution of size, zeta potential, density of fluorescently tagged surface markers, and nucleic acid cargo, along with the ability to isolate/collect specific EV subpopulations with tightly defined biophysical parameters, all on a single platform. The feasibility of the proposed system will be demonstrated by building a prototype platform and showing its ability to characterize, sort and isolate EV samples. The resulting system will have an ease-of-use, throughput, and price point similar to flow cytometry, dynamic light scattering (DLS), nanoparticle tracking analysis (NTA), or single-pass resistive-pulse techniques, with a resolution comparable to microscopy techniques, e.g. scanning or tunneling electron microscopy (SEM or TEM, respectively) or atomic force microscopy (AFM), yielding a state-of-the-art instrument which will directly enable and advance the field of EV diagnostics.