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

基本信息

批准号：
9982238
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
项目类别：
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/9982238
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 In Vitro 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 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

项目摘要

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.

细胞外蔬菜（EV）是一类新的循环生物标志物，有望无创，实时癌症监测。许多研究表明，i）电动汽车可能充当父母癌的可靠替代物细胞和ii）这些蔬菜可以反映全球肿瘤伯恩，克服了肿瘤异质性和采样偏见。然而，将电动汽车分析转化为临床相关的癌症测试受到缺乏的限制标准化的实用方法。常规测定工具（例如，超速离心，蛋白质印迹，ELISA）需要大量的电动汽车和广泛的处理，使其对于临床工作流动不切实际。此外，样本处理和测试协议的变化通常会导致不一致和混淆结果。该提案的目的是i）解决电动汽车分析中的此类技术挑战，ii）严格评估EVS的临床价值作为早期检测卵巢癌的新生物标志物。我们制定了一个策略实现这一目标的学术行业伙伴关系：外泌体诊断，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互补资源。首先，我们会的推进标准化的EV分析平台。我们将采用我们最近开发的放电平台流线电动汽车收集和高通量EV蛋白的NPLEX（纳米质量外部）技术筛选。我们的最初研究表明，NPLE的灵敏度比常规的灵敏度高1000倍方法但却消耗了少量样品体积（0.1μl）。我们现在寻求将NPLE推向乐器 i）级别i）提高其鲁棒性和吞吐量，ii）建立标准化的测定协议。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，特别是测试是否可以探索电动汽车作为早期发现卵巢癌和的生物标志物进程监测。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遗传学设计的小鼠模型以鉴定卵巢癌的EV蛋白特征。我们接下来会流通来自卵巢癌患者的电动汽车，并评估肿瘤燃烧与EV蛋白质签名之间的相关性。我们的研究将旨在确保评估可靠性和可重复性，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施加严格的质量控制，积累良好的患者和对照样品，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的翻译影响，建立对早期检测卵巢癌的强大，高度特异性的活检。