Rapid Protease Profiling with a Multiplex Electronic Method for Detection of Metastatic Triple-Negative Breast Cancer

使用多重电子方法快速进行蛋白酶分析，检测转移性三阴性乳腺癌

• 批准号: 9355398
• 负责人: JUN LI
• 金额: $ 40.01万
• 依托单位: KANSAS STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9355398
• 关键词: Address; Algorithms; Basement membrane; Biological Assay; Biological Markers; Biological Response Modifier Therapy; Biomedical Engineering; Biosensor; Blood Circulation; Blood specimen; Body Fluids; Brain; Buffers; Caliber; Cancer Patient; Carbon; Carbon Nanotubes; Cathepsins B; Cells; Cessation of life; Clinical; Clinical Research; Collaborations; Complex; Data; Detection; Development; Diagnosis; Diagnostic; Discipline; Disease; Disintegrins; Distal; Distant; Drug Targeting; ERBB2 gene; Early Diagnosis; Electronics; Enzymes; Estrogens; Extracellular Matrix; Extravasation; Frequencies; Gelatinase A; Gelatinase B; Goals; Hydrolysis; Individual; Kansas; Kinetics; Length; Link; Liquid substance; Liver; Lung; Lymph; Malignant Neoplasms; Mammary Gland Parenchyma; Matrix Metalloproteinases; Measures; Medical center; Metalloproteases; Metastatic breast cancer; Methods; Monitor; Nanotechnology; Neoplasm Metastasis; Oncologist; Operative Surgical Procedures; Oxidation-Reduction; Patients; Pattern; Peptide Hydrolases; Peptide Library; Peptides; Pharmaceutical Chemistry; Physiological Processes; Positive Lymph Node; Preclinical Drug Evaluation; Progesterone; Protease Inhibitor; Proteins; Radiation; Recurrence; Reporter; Reporting; Research; Research Project Grants; Role; Sampling; Savings; Sensitivity and Specificity; Serum; Signal Transduction; Speed; Statistical Data Interpretation; Surface; System; TNF-alpha converting enzyme; Techniques; Technology; Therapeutic; Time; Tissues; Tumor Cell Invasion; United States National Aeronautics and Space Administration; Universities; Urokinase; base; bone; breast cancer diagnosis; cancer biomarkers; cancer cell; cancer diagnosis; cancer invasiveness; cancer recurrence; cancer therapy; cancer type; chemotherapy; clinical biomarkers; cost effective; cysteine rich protein; design; effective therapy; enzyme activity; extracellular; ferrocene; follow-up; high risk; hormone therapy; inhibitor/antagonist; lymph nodes; malignant breast neoplasm; nanodevice; nanoelectronics; nanofiber; nanomolar; neoplastic cell; overexpression; portability; rapid technique; screening; tool; treatment response; triple-negative invasive breast carcinoma; tumor progression

Early diagnosis via cost-effective screening of high risk subpopulation followed by effective treatments is the key for saving many lives of cancer patients. Proteases overexpressed in cancer cells and secreted into circulation have been used as drug targets for the development of protease inhibitors as well as biomarkers for diagnosis and therapeutic monitoring. In this project, a multi-discipline team intends to develop a nanoelectronic chip for rapid profiling activity of six proteases that are overexpressed in metastatic triple-negative breast cancer (TNBC) patients and use this information to identify signatures in breast cancer progression and therapeutic responsiveness. The team includes (1) Dr. Jun Li (PI, nanotechnology and biosensor, Kansas State University); (2) Dr. Duy H. Hua (synthetic medicinal chemistry, Kansas State University); (3) Dr. Meyya Meyyappan (nanotechnology and nanodevices, NASA Ames Research Center); and (4) Dr. Priyanka Sharma (cancer clinical research, the University of Kansas Medical Center). This project focuses on developing the PI’s previously demonstrated electronic device based on carbon nanofiber nanoelectrode arrays for simultaneous detection of the activity of six overexpressed proteases in TNBC, including cathepsin B, MMP-2, MMP-9, ADAM-10, ADAM- 17 and uPA. Hexapeptide substrates highly specific to the aforementioned proteases are covalently attached to the tips of embedded carbon nanofibers in a multiplexed electronic chip. The distal end of the hexapeptides is attached with an electrochemical reporter. Introducing the TNBC patient samples containing these proteases results in cleavage of corresponding hexapeptides, thereby releasing the electrochemical reporter and leading to an exponential decrease of the electrochemical signal. The protease activity can be derived from the decay time constant of the kinetic curves. This extremely sensitive biosensor technology will be fabricated into independently addressed 3x3 arrays and packaged in disposable fluidic cartridges. Using 18 hexapeptide substrates (3 for each protease) attached to different nanoelectrode arrays in two 3x3 electronic chips, reliable proteolytic activity profile of the aforementioned six proteases in TNBC can be quickly measured, which cannot be done with current technologies. This technique should greatly accelerate the speed of protease profiling in complex samples and facilitate detection of invasive TNBC from other breast cancers in its early stage. The change of the protease activity profile will be monitored and correlate it with the longitudinal cancer treatment responses.

通过对高风险亚群的成本效益筛查，然后有效的早期诊断 治疗是挽救许多癌症患者生命的关键。蛋白酶过表达 癌细胞和分泌成循环已被用作发育的药物靶标 蛋白质抑制剂以及用于诊断和治疗监测的生物标志物。在这个 项目是一个多学科团队，打算开发纳米电子芯片以快速分析 在转移性三阴性乳腺癌中过表达的六种蛋白酶的活性 （TNBC）患者并使用此信息来识别乳腺癌进展和 治疗反应能力。该团队包括（1）Jun Li博士（PI，纳米技术和 堪萨斯州立大学生物传感器）； （2）Duy H. Hua博士（堪萨斯州的合成药物化学 州立大学）; （3）Meyya Meyyappan博士（NASA AMES的纳米技术和纳米技术 研究中心）； （4）Priyanka Sharma博士（癌症临床研究，大学 堪萨斯医疗中心）。 该项目着重于开发PI先前演示的电子设备的基于 碳纳米纤维纳米电极阵列，以简单检测六个活性 TNBC中过表达的蛋白酶，包括组织蛋白酶B，MMP-2，MMP-9，Adam-10，Adam- 17和UPA。六磷酸底物高度特异性近似蛋白酶为 共价附着在多路复用电子芯片中的嵌入碳纳米纤维的尖端上。 六肽的远端与电化学记者相连。介绍 包含这些蛋白酶的TNBC患者样品导致相应的裂解 六肽，从而释放电化学记者并导致指数 电化学信号的减少。蛋白酶活性可以从衰减中得出 动力学曲线的时间常数。 这种非常敏感的生物传感器技术将被制造为独立解决 3x3阵列并包装在一次性液体墨盒中。使用18个六磷酸底物（3 对于每个蛋白酶））在两个3x3电子芯片中附着在不同的纳米电极阵列上 TNBC中近似六个蛋白酶的可靠蛋白水解活性谱可以很快 测量，目前的技术无法完成。这项技术应该很大 加速复杂样品中蛋白酶分析的速度，并促进检测 在早期的其他乳腺癌中，来自其他乳腺癌的侵入性TNBC。蛋白酶活动的变化 剖面将受到监测并将其与纵向癌症治疗反应相关联。