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

基本信息

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
项目类别：
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

项目摘要

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.

通过对高风险亚群进行具有成本效益的筛查，然后进行有效的早期诊断治疗是挽救许多癌症患者生命的关键。癌细胞并分泌到循环中已被用作开发的药物靶标蛋白酶抑制剂以及用于诊断和治疗监测的生物标志物。项目中，一个多学科团队打算开发一种用于快速分析的纳米电子芯片在转移性三阴性乳腺癌中过度表达的六种蛋白酶的活性 (TNBC) 患者，并使用这些信息来识别乳腺癌进展的特征和该团队包括（1）李军博士（PI，纳米技术和生物传感器，堪萨斯州立大学）；（2）Duy H. Hua 博士（合成药物化学，堪萨斯州） (3) Meyya Meyyappan 博士（纳米技术和纳米器件，美国宇航局艾姆斯研究中心 (4) Priyanka Sharma 博士（癌症临床研究，英国大学）堪萨斯医疗中心）。该项目的重点是开发 PI 先前展示的基于碳纳米纤维纳米电极阵列可同时检测六个活性 TNBC 中过度表达的蛋白酶，包括组织蛋白酶 B、MMP-2、MMP-9、ADAM-10、ADAM- 17 和 uPA 底物对上述蛋白酶具有高度特异性。共价连接到多路电子芯片中嵌入的碳纳米纤维的尖端。六肽的远端连接有电化学报告基因。含有这些蛋白酶的 TNBC 患者样本会导致相应的蛋白酶裂解六肽，从而释放电化学报告基因并导致指数电化学信号的降低可以源自蛋白酶活性的衰减。动力学曲线的时间常数。这种极其灵敏的生物传感器技术将被制造成独立寻址的 3x3 阵列并包装在一次性流体盒中，使用 18 个六肽底物 (3)。对于每种蛋白酶）连接到两个 3x3 电子芯片中的不同纳米电极阵列，可以快速获得 TNBC 中提到的六种蛋白酶的可靠蛋白水解活性谱测量，这是目前技术无法做到的。加快复杂样品中蛋白酶分析的速度并促进检测其他乳腺癌早期侵袭性 TNBC 蛋白酶活性的变化。将监测概况并将其与纵向癌症治疗反应相关联。