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（EVs）是一类新的循环生物标志物，有望实现非侵入性，实时 癌症监测许多研究表明，i）EV可能是父母癌症的可靠替代品 ii）这些囊泡可以反映整体肿瘤负荷，克服肿瘤异质性的限制， 抽样偏差然而，将EV分析转化为临床相关的癌症测试受到缺乏 标准化、实用化的方法。常规的化验工具（例如，蛋白质印迹法，ELISA） 需要大量的EV和大量的处理，使得它们对于临床工作流程来说是不切实际的。 此外，样品处理和测试方案的变化通常导致不一致和混淆。 结果本提案的目标是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生物标志物的临床价值。我们建立了一个战略 为实现这一目标，学术界与工业界建立了合作伙伴关系：Exosome Diagnostics，一家基于电动汽车的行业领导者， 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的系统生物学中心， 用于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 技术和补充资源，以执行以下转化项目。一是 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， 流线型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的灵敏度比传统的高出1000倍以上。 方法，但消耗少量样品体积（0.1 μL）。我们现在寻求将nPLEX推进到仪器中 通过i）提高其鲁棒性和通量，以及ii）建立标准化的测定方案来提高水平。 利用外泌体诊断的开发和监管专业知识，由此产生的平台将 准备转化为临床诊断实验室。第二，我们将进行有针对性的临床研究， 特别是测试EV是否可以用作早期检测卵巢癌的生物标志物， 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的遗传学方法。 工程小鼠模型，以确定卵巢癌的EV蛋白特征。我们将在下一个配置文件循环 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检测的可靠性和重现性，从而在临床上提供 可翻译的EV诊断。我们将对设备设计和样品处理实施严格的质量控制， 收集注释良好的患者和对照样本，并进行多点检测。技术和科学 这项研究的结果可能对癌症管理产生重大的转化影响， 一个强大的，高度特异性的液体活检，用于卵巢癌的早期检测。