Exosome-Protein-microRNA-OneStop (Exo-PROS) biosensor: a new liquid biopsy for cancer screening and early detection

外泌体-蛋白质-microRNA-OneStop (Exo-PROS) 生物传感器：用于癌症筛查和早期检测的新型液体活检

• 批准号: 10115649
• 负责人: Qiaoqiang Gan
• 金额: $ 20.64万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10115649
• 关键词: A549; Address; Advanced Development; Affect; Antibodies; Benign; Biological Assay; Biological Markers; Biometry; Biosensor; Buffaloes; Cancer Diagnostics; Cancer Patient; Cell Culture Techniques; Cells; Clinical; Colonoscopy; Colorectal Cancer; Complement; Complementary DNA; Consumption; Culture Media; Data; Detection; Development; Devices; Diagnosis; Diagnostic; Diagnostic tests; Disease model; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor Receptor; Fluorescence; Goals; Hour; Immunomagnetic Separation; Knowledge; Malignant Neoplasms; Malignant neoplasm of lung; Measures; Medical Imaging; Membrane Proteins; Methods; MicroRNAs; Molecular; Monitor; Names; Neoplasm Circulating Cells; Non-Small-Cell Lung Carcinoma; Normal Cell; Oncology; Optics; Patients; Performance; Pilot Projects; Portraits; Procedures; Prognosis; Proteins; Pulmonary Coin Lesion; Quantitative Reverse Transcriptase PCR; RNA; Research Personnel; Risk Factors; Roswell Park Cancer Institute; Sampling; Screening for cancer; Sensitivity and Specificity; Serum; Signal Transduction; Smoker; Surface Plasmon Resonance; Surface Properties; System; Techniques; Technology; Testing; Time; Tumor-Derived; Validation; base; biobank; biochip; biomarker discovery; cancer biomarkers; cancer cell; cancer diagnosis; circulating biomarkers; cohort; cost; cost effective; design; detection limit; detection sensitivity; diagnostic accuracy; exosome; experience; high risk; improved; in-vitro diagnostics; liquid biopsy; low dose computed tomography; lung cancer screening; microRNA biomarkers; multidisciplinary; nanophotonic; next generation sequencing; overexpression; personalized medicine; plasmonics; protein biomarkers; screening; treatment response; tumor; tumor DNA

PROJECT SUMMARY Current cancer screening tests, such as low dose computed tomography (LDCT) for lung cancer and colonoscopy for colorectal cancer, are challenged by low sensitivity, high false positive rate, limited tumor information, uncomfortable or invasive procedures and high cost. Liquid biopsy that detects circulating biomarkers, allows sequential monitoring of cancer development, complements medical imaging and risk factor data to provide a more comprehensive portrait of cancer, and thus it has shown great promise as the in vitro diagnostics for cancer. Exosomal proteins and exosomal microRNAs are promising cancer biomarkers for liquid biopsy, and have shown very high sensitivity and specificity in diagnosing many cancers. Unfortunately current detection technologies, such as ELISA, LC-MS, qRT-PCR, microarray and next generation sequencing, are tedious, expensive and time consuming, which limits their clinical utilities as screening tests. Moreover, many methods cannot distinguish tumor-derived exosomes from their non-tumor counterparts, and thus suffer from poor detection sensitivity and specificity. To address these challenges, we have developed a surface plasmon resonance (SPR)-based, molecular beacon-assisted liquid biopsy device, named Exosome Protein microRNA OneStop (Exo-PROS) biosensor. This biosensor can capture tumor-derived exosomes using cancer-overexpressed proteins. It can also provide one-stop quantification of surface proteins and intra- vesicular microRNAs of captured exosomes on a single device within 4 hours. The effective separation of tumor-derived-exosomes from interfering exosomes (such as normal-cell-derived exosomes) and the combination of exosomal protein and microRNA biomarkers may significantly increase the diagnostic accuracy. We have successfully used the Exo-PROS biosensor to measure exosomal EGFR and EGFR+ exosomal miR- 21 in cell culture medium and lung cancer patient serum samples. The biosensor showed higher sensitivity and specificity than ELISA and qRT-PCR in distinguishing cancer from normal controls. In this study, we propose to (1) advance the development of the Exo-PROS biosensor to improve its sensing performance and to realize multiplexed biomarker detection; (2) demonstrate its clinical utility in lung cancer screening and early detection, and explore the diagnostic value of exosomal protein and microRNA combined biomarkers in cancer, which has not yet been investigated. We will evaluate the detection sensitivity and specificity of Exo-PROS biosensor in distinguishing nonsmall cell lung cancer patients from healthy controls and smokers with benign solitary pulmonary nodule at high risk for lung cancer. We will demonstrate the superior sensing performance of Exo- PROS biosensor over ELISA and qRT-PCR assays. We expect to develop the Exo-PROS biosensor into a highly accurate, simple, fast, and cost-effective liquid biopsy test for cancer screening and early detection.

项目摘要 当前的癌症筛查测试，例如用于肺癌和 结肠癌的结肠镜检查受到低灵敏度，高误报率，有限的肿瘤的挑战 信息，不舒服或侵入性程序以及高成本。检测循环的液体活检 生物标志物，允许对癌症发展的顺序监测，补充医学成像和危险因素 数据提供更全面的癌症肖像，因此，它表现出了巨大的希望 癌症的诊断。外泌体蛋白和外泌体microRNA是有希望的癌症生物标志物 液体活检，并且在诊断许多癌症时表现出很高的灵敏度和特异性。很遗憾 当前的检测技术，例如ELISA，LC-MS，QRT-PCR，微阵列和下一代测序， 乏味，昂贵且耗时，这限制了其临床公用事业作为筛查测试。而且， 许多方法无法将肿瘤衍生的外泌体与非肿瘤的外泌体区分开，因此 从较差的检测灵敏度和特异性中。为了应对这些挑战，我们已经发展了一个表面 基于等离子共振（SPR）的分子信标辅助液体活检装置，称为外泌体蛋白 microRNA oneStop（Exo-pros）生物传感器。该生物传感器可以使用 癌症过表达的蛋白质。它还可以提供表面蛋白和内部表面蛋白质的一站式定量 在4小时内，在单个设备上捕获的外泌体的水泡microRNA。有效的分离 来自干扰外泌体（例如正常细胞衍生的外泌体）和肿瘤衍生的外观 外泌体蛋白质和microRNA生物标志物的组合可能会显着提高诊断准确性。 我们已经成功使用Exo-Pros生物传感器来测量外泌体EGFR和EGFR+外泌体miR- 21在细胞培养基和肺癌患者血清样品中。生物传感器显示较高的灵敏度，并且 比ELISA和QRT-PCR的特异性在区分癌症和正常对照方面的特异性。在这项研究中，我们提出 （1）推进Exo-Pros生物传感器的开发，以提高其感应性能并实现 多路复用生物标志物检测； （2）证明其在肺癌筛查和早期检测中的临床实用性 并探索外泌体蛋白质和microRNA在癌症中合并生物标志物的诊断价值，这 尚未进行调查。我们将评估EXO-PROS生物传感器的检测灵敏度和特异性 在区分非健康对照者和良性孤立的吸烟者时，将非小细胞肺癌患者区分开 肺癌的肺结节高风险。我们将展示Exo-exo-的卓越感应性能 对ELISA和QRT-PCR分析的优点生物传感器。我们希望将Exo-Pros生物传感器发展为 高度准确，简单，快速且具有成本效益的液体活检测试，用于癌症筛查和早期检测。