Clinical platform for high-throughput analyses of extracellular vesicles

细胞外囊泡高通量分析的临床平台

• 批准号: 10224771
• 负责人: Hakho Lee
• 金额: $ 58.88万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 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/10224771
• 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; 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; diagnostic technologies; exosome; extracellular vesicles; high throughput analysis; in-vitro diagnostics; 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

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.

细胞外蔬菜（EV）已成为组织活检的诺言替代 实现无创，实时的癌症监测。大多数癌细胞将大量电动汽车释放到 携带分子的循环构成父肿瘤。因此，分析电动汽车可能会提供新的途径 评估肿瘤烧伤和肿瘤异质性。但是，通过将电动汽车分析用于临床工作流程 由于缺乏标准化的实用方法而阻碍。常规评估工具（例如，超速离心， Western印迹，ELISA）需要大量样品和广泛的处理；他们对 临床应用。样品处理和测试协议的变化通常会导致不一致和可变 发现。该提案的目的是i）通过推进一个强大的平台来应对此类技术挑战 EV蛋白分析，ii）严格评估EVS作为癌症生物标志物的临床价值。我们形成了一个 战略学术行业伙伴关系以实现这一目标：外部诊断，行业领导者 基于EV的癌症诊断，提供了开发和制造体外诊断医学的现成能力 设备；以及马萨诸塞州综合医院的系统生物学中心，这是发展新颖的先驱 EV分析的分析技术。这些团队将带来他们的多学科专业知识，创新的 进行以下翻译项目的技术和完成资源：首先，我们将建立 一个敏感的高通量平台，用于EV蛋白筛选。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（纳米质量外显子）技术 通过纳米结构。我们的初步研究表明，NPLE的灵敏度比 常规方法但却消耗了很少的样品体积（0.1μl）。新的nplex系统将有 商业生产的分析能力和可伸缩性扩大。我们将设计一个新的SPR芯片和一个 检测工具，用于大规模并行EV筛选，还建立了标准化的测定方案。 利用外泌体诊断的发展和监管专业知识，由此产生的平台将是 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 治疗监测。我们将从接受疗法的卵巢癌患者那里收集循环电动汽车，并 跟踪EV蛋白水平的串行变化。我们将确保分析可靠性和可重复性 临床翻译的电动汽车诊断。我们将在设备设计和样品上遵循严格的质量控制 加工，累积良好的患者和对照样品，并进行多站点测试。技术 这项研究的科学结果可能对癌症产生重大的翻译影响，建立 强大的，高度具体的评估，以指导治疗决策和评估疗法效率。外泌体 诊断将提供监管指导以及销售新测试和设备的渠道。