Clinical platform for high-throughput analyses of extracellular vesicles

细胞外囊泡高通量分析的临床平台

基本信息

批准号：
9754806
负责人：
Hakho Lee
金额：
$ 56.51万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-02 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9754806
关键词：
Address Adopted Adoption Benchmarking Biological Assay Biomedical Engineering Biopsy Biotechnology Blood Circulation Blood specimen Cancer Detection Cancer Diagnostics Cancer Patient Cells Clinic Clinical Clinical Research Consumption Dana-Farber Cancer Institute Detection Development Device Designs Devices Diagnosis Diagnostic Disease Drug resistance Enrollment Ensure Enzyme-Linked Immunosorbent Assay Evolution General Hospitals Goals Gold Heterogeneity Immunotherapeutic agent In Vitro Industry Industry Standard Laboratories Lead Malignant Neoplasms Malignant neoplasm of ovary Massachusetts Measures Medical Device Methods Microfabrication Molecular Molecular Analysis Monitor Nature Nucleic Acids Outcomes Research Parents Patients Periodicity Pharmaceutical Preparations Platinum Preparation Production Protein Analysis Proteins Protocols documentation Quality Control RNA Reader Reproducibility Research Resources Sampling Screening for Ovarian Cancer Signal Transduction Standardization Structure Surface Plasmon Resonance System Systems Biology Techniques Technology Testing Therapy trial Thinking Time Tissues Translating Translations Treatment Efficacy Treatment outcome Tumor Burden Ultracentrifugation Variant Vesicle Western Blotting base cancer biomarkers cancer care cancer cell cancer drug resistance cancer therapy circulating biomarkers clinical application clinical diagnostics clinical translation clinically translatable design exosome extracellular vesicles high throughput analysis industry partner inhibitor/antagonist innovation innovative technologies instrument liquid biopsy multidisciplinary nanoplasmonic next generation novel novel diagnostics patient oriented research personalized cancer therapy precision medicine screening sensor technology success tool translational impact treatment response tumor tumor heterogeneity

项目摘要

Extracellular vesicles (EVs) have emerged as a promising surrogate for the tissue biopsy, potentially enabling non-invasive, real-time cancer monitoring. Most cancer cells release large numbers of EVs into circulation that carry molecular constituents of the parent tumor. Analyzing EVs could thus offer new avenues to assess tumor burden and tumor heterogeneity. Adoption of EV analyses into a clinical workﬂow, however, is impeded by the lack of standardized, practical methods. Conventional assay tools (e.g., ultracentrifugation, Western blotting, ELISA) require large sample volumes and extensive processing; they are impractical for clinical applications. Variations in sample handling and testing protocols often lead to inconsistent and variable ﬁndings. The goal of this proposal is i) to address such technical challenges by advancing a robust platform for EV protein analyses, and ii) to rigorously evaluate EVs' clinical value as cancer biomarkers. We formed a strategic academic-industry partnership to achieve this goal: Exosome Diagnostics, an industry leader in EV-based cancer diagnostics, offering ready capacity to develop and manufacture in-vitro diagnostic medical devices; and the Center for Systems Biology at Massachusetts General Hospital, a pioneer in developing novel analytical technologies for EV analyses. These teams will bring in their multidisciplinary expertise, innovative technologies and complementary resources to carry out the following translational projects: First, we will build a sensitive, high-throughput platform for EV protein screening. The system will adopt our recently developed nPLEX (nano-plasmonic exosome) technology that is based on novel surface plasmon resonance (SPR) through nanohole structures. Our initial study showed that nPLEX achieved >1000-fold higher sensitivity than conventional methods and yet consumed scant sample volumes (0.1 μL). The new nPLEX system will have expanded analytical capacity and scalability for commercial production. We will design a new SPR chip and a detection instrument for massively parallel EV screening, and also establish standardized assay protocols. Leveraging the developmental and regulatory expertise of Exosome Diagnostics, the resulting platform will be ready for translation into clinical diagnostic laboratories. Second, we will perform a targeted clinical study, particularly testing whether EV protein signatures can be used as a biomarker for cancer detection and treatment monitoring. We will collect circulating EVs from ovarian cancer patients undergoing therapies, and track serial changes of EV protein levels. We will ensure assay reliability and reproducibility to deliver clinically translatable EV diagnostics. We will follow stringent quality control on device design and sample processing, accrue well-annotated patient and control samples, and perform multisite testing. The technical and scientiﬁc outcomes of this research could have a signiﬁcant translational impact in cancer, establishing a robust, highly speciﬁc assay to guide treatment decision and assess therapeutic efﬁcacy. Exosome Diagnostics will provide regulatory guidance as well as channels to market new tests and devices.

细胞外囊泡（EV）已成为组织活检的有前途的替代品，有可能实现非侵入性实时癌症监测大多数癌细胞会释放大量 EV。因此，分析携带母体肿瘤分子成分的循环可以提供新的途径。然而，将 EV 分析纳入临床工作流程来评估肿瘤负荷和肿瘤异质性是必要的。由于缺乏标准化、实用的方法（例如超速离心、蛋白质印迹（ELISA）需要大量样本和大量处理，因此不切实际；临床应用中样品处理和测试方案的变化常常导致不一致和可变。该提案的目标是 i) 通过推进一个强大的平台来解决此类技术挑战。 EV 蛋白分析，以及 ii) 严格评估 EV 作为癌症生物标志物的临床价值。学术与产业战略合作伙伴关系以实现这一目标：Exosome Diagnostics，行业领导者基于 EV 的癌症诊断，提供开发和制造体外诊断医疗的现成能力设备；以及马萨诸塞州总医院系统生物学中心，该中心是开发新型药物的先驱这些团队将带来他们的多学科专业知识和创新的分析技术。技术和互补资源来开展以下转化项目：首先，我们将建立用于EV蛋白筛选的灵敏、高通量平台该系统将采用我们最近开发的。基于新型表面等离子体共振 (SPR) 的 nPLEX（纳米等离子体外泌体）技术我们的初步研究表明，nPLEX 的灵敏度比纳米孔高 1000 倍以上。传统方法，但消耗的样品量很少（0.1 µL），新的 nPLEX 系统将具有。扩大了商业生产的分析能力和可扩展性，我们将设计一种新的SPR芯片和一个大规模并行 EV 筛查的检测仪器，并建立标准化的检测方案。利用外泌体诊断的开发和监管专业知识，所得平台将是准备转化为临床诊断实验室其次，我们将进行有针对性的临床研究，特别是测试 EV 蛋白特征是否可以用作癌症检测的生物标志物我们将收集接受治疗的卵巢癌患者的循环 EV。我们将跟踪 EV 蛋白水平的连续变化，以确保检测的可靠性和重现性。我们将在设备设计和样品方面遵循严格的质量控制。处理、积累注释良好的患者和对照样本，并执行多站点测试。这项研究的科学成果可能对癌症产生重大的转化影响，建立一种强大的、高度特异性的检测方法，用于指导治疗决策和评估治疗效果。诊断将提供监管指导以及新测试和设备的营销渠道。