Early Detection through Novel OCEAN Technology - Ovarian Cancer Exosomal Analysis with Nanoplasmonics

通过新型 OCEAN 技术进行早期检测 - 使用纳米等离子体技术进行卵巢癌外泌体分析

• 批准号: 10223236
• 负责人: Cesar M Castro
• 金额: $ 74.67万
• 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: 2018-09-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10223236
• 关键词: Address; Adopted; Adoption; Animal Model; Ascites; Benign; Biological; Biological Assay; Blood Tests; Cancer Diagnostics; Cancer Patient; Carcinoma; Cell Culture Techniques; Cells; Clinical; Clinical Research; Collection; Complex; Consumption; Culture Media; Custom; Development; Device Designs; Diagnostic; Early Diagnosis; Ensure; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Epithelial ovarian cancer; Excision; General Hospitals; Genetic Risk; Genetically Engineered Mouse; Goals; Hospitals; Human; Industry; Industry Standard; Laboratories; Lead; Liquid substance; Malignant - descriptor; Malignant Female Reproductive System Neoplasm; Malignant Neoplasms; Malignant neoplasm of ovary; Mammalian Oviducts; Massachusetts; Measures; Medical Device; Membrane; Membrane Proteins; Methods; Microfabrication; Modeling; Molecular; Molecular Analysis; Monitor; Mutation; Nanotechnology; Nucleic Acids; Oceans; Operative Surgical Procedures; Outcomes Research; Ovarian; Ovary; Pathogenesis; Pathology; Patients; Performance; Perinatal; Peritoneal; Plasma; Preclinical Testing; Preparation; Protein Analysis; Proteins; Protocols documentation; Quality Control; Reader; Reproducibility; Resources; Sampling; Sampling Biases; Screening for Ovarian Cancer; Screening for cancer; Serous; Signal Transduction; Standardization; Statistical Data Interpretation; System; Systems Biology; TP53 gene; Techniques; Technology; Testing; Thinking; Time; Translating; Translations; Tumor Burden; Ultracentrifugation; Validation; Variant; Vesicle; Western Blotting; Woman; advanced disease; assay development; base; cancer care; cancer cell; circulating biomarkers; clinical diagnostics; clinical translation; clinically relevant; clinically translatable; comparative; design; early detection biomarkers; exosome; experience; extracellular vesicles; high risk population; high throughput technology; improved; in-vitro diagnostics; industry partner; innovation; innovative technologies; instrument; laboratory equipment; liquid biopsy; minimally invasive; multidisciplinary; nanoplasmonic; nanoscale; next generation; novel; novel marker; particle; peripheral blood; pre-clinical research; preclinical study; product development; prophylactic; prospective; research clinical testing; screening; sensor; sensor technology; success; survivorship; tool; translational impact; tumor heterogeneity; tumor progression

Extracellular vesicles (EVs) are a new class of circulating biomarker that promises non-invasive, real-time cancer monitoring. Many studies have shown that i) EVs may function as reliable surrogates of parental cancer cells, and ii) these vesicles can reflect global tumor burden, overcoming limitations of tumor heterogeneity and sampling bias. Translating EV analyses into a clinically relevant cancer test, however, is limited by the lack of standardized, practical methods. Conventional assay tools (e.g., ultracentrifugation, Western blotting, ELISA) require large amounts of EVs and extensive processing, making them impractical for clinical workflows. Moreover, variations in sample handling and testing protocols often lead to inconsistent and confounding results. The goal of this proposal is to i) address such technical challenges in EV analyses, and ii) rigorously evaluate EVs' clinical value as a novel biomarker for early detection of ovarian cancer. 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 the Massachusetts General Hospital, a pioneer in developing novel analytical technologies for EV analyses. These teams will bring their multidisciplinary expertise, innovative technologies and complementary resources to carry out the following translational projects. First, we will advance a standardized EV assay platform. We will adopt our recently developed ExoLution platform to streamline EV collection, and the nPLEX (nano-plasmonic exosome) technology for high-throughput EV protein screening. Our initial study showed that nPLEX achieved >1000-fold higher sensitivity than conventional methods and yet consumed scant sample volumes (0.1 μL). We now seek to advance nPLEX to the instrument level by i) improving its robustness and throughput, and ii) establishing 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 EVs can be exploited as a biomarker for early detection of ovarian cancer and progression monitoring. Our preclinical study will use patient-derived ovarian cancer cells and novel genetically engineered mouse models to identify EV protein signatures for ovarian cancer. We will next profile circulating EVs from ovarian cancer patients and assess the correlation between tumor burden and EV protein signature. Our study will be designed to ensure assay reliability and reproducibility, thereby delivering clinically translatable EV diagnostics. We will impose stringent quality controls 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 management, establishing a robust, highly specific liquid biopsy for early detection of ovarian cancer.

细胞外囊泡 (EV) 是一类新型循环生物标志物，有望实现非侵入性、实时检测 癌症监测。许多研究表明 i) EV 可能作为亲本癌症的可靠替代物 细胞，ii）这些囊泡可以反映整体肿瘤负荷，克服肿瘤异质性的局限性和 抽样偏差。然而，将 EV 分析转化为临床相关的癌症测试由于缺乏 标准化、实用的方法。传统检测工具（例如超速离心、蛋白质印迹、ELISA） 需要大量的 EV 和广泛的处理，这使得它们对于临床工作流程来说不切实际。 此外，样品处理和测试协议的变化常常导致不一致和令人困惑的结果 结果。该提案的目标是 i) 解决 EV 分析中的此类技术挑战，以及 ii) 严格 评估 EV 作为卵巢癌早期检测的新型生物标志物的临床价值。我们形成了战略 学术与行业合作以实现这一目标：Exosome Diagnostics，基于 EV 的行业领导者 癌症诊断，提供开发和制造体外诊断医疗设备的现成能力； 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 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、创新能力 技术和补充资源来开展以下转化项目。首先，我们将 推进标准化 EV 检测平台。我们将采用我们最近开发的 ExoLution 平台来 简化 EV 收集，以及用于高通量 EV 蛋白的 nPLEX（纳米等离子体外泌体）技术 筛选。我们的初步研究表明，nPLEX 的灵敏度比传统方法高 1000 倍以上 方法，但消耗的样品量很少（0.1 μL）。我们现在寻求将 nPLEX 推进到仪器中 通过 i) 提高其稳健性和通量，以及 ii) 建立标准化测定协议来提高水平。 利用外泌体诊断的开发和监管专业知识，所得平台将是 准备好转化为临床诊断实验室。其次，我们会进行有针对性的临床研究， 特别是测试 EV 是否可以用作卵巢癌早期检测的生物标志物 进展监测。我们的临床前研究将使用患者来源的卵巢癌细胞和新颖的基因 设计小鼠模型来识别卵巢癌的 EV 蛋白特征。我们将在接下来的简介中流传 来自卵巢癌患者的 EV 并评估肿瘤负荷和 EV 蛋白特征之间的相关性。 我们的研究旨在确保测定的可靠性和重现性，从而提供临床 可翻译的电动汽车诊断。我们将对器件设计和样品加工进行严格的质量控制， 积累注释良好的患者和对照样本，并进行多站点测试。技术和科学 这项研究的结果可能对癌症管理产生重大转化影响，建立 用于早期检测卵巢癌的强大、高度特异性的液体活检。