Solid-state nanopore-based MMP/ADAM profiling for early cancer detection

非技术性：该奖项由材料研究部的生物材料计划授予伊利诺伊理工学院，旨在开发基于两组生物催化剂的下一代生物材料，即基质金属蛋白酶(MMPs)和亚当斯(一种去整合素和金属蛋白酶)，用于传感和其他技术中的可能应用。MMPs和ADAMS是负责细胞外基质降解和组织重塑的酶，在各种生物、生理和病理过程中发挥重要作用。它们目前正在进行深入的研究，作为新的生物标志物和潜在的治疗靶点，用于早期发现和可能的人类癌症治疗。然而，目前的基质金属蛋白酶/ADAM检测方法存在特异性低、选择性差等缺点，限制了其在临床上的应用。本项目提供了一种新的无标记策略，将纳米孔传感和基于底物的蛋白酶检测相结合，实现了对MMPs/ADAMS活性的高选择性和高灵敏度的测量。预计拟议的研究将有助于更好地了解分子和离子传输，并开发一种多功能工具，用于各种应用，包括生物传感、研究共价和非共价键相互作用、研究生物分子折叠和去折叠，以及探索酶动力学。除研究外，教育和宣传部分也是该项目的组成部分。来自这项工作的最新研究成果将被纳入课堂教学，以造福于没有直接参与这项研究的学生。PI将通过既定的当地项目招募本科生和研究生(特别是西班牙裔和非裔美国少数群体)参与研究。国际和平研究所将通过组织跨学科研讨会来传播纳米孔传感技术，以促进思想交流，促进同行和普通公众之间的国内和国际合作。技术：该项目将探索一种新的策略来测量MMPs(基质金属蛋白酶)和ADAMS(一种去整合素和金属蛋白酶)的活性，这是通过实时监测由纳米孔中的蛋白酶-底物肽相互作用引起的离子电流调制来实现的。通过利用底物的特异性和底物的裂解位点，纳米孔传感器提供了显著提高MMPs/ADAMS分析的选择性和准确性的潜力。该项目的三个目标是：1)研究影响多肽在纳米通道中传输的各种因素，并了解其潜在的机制；2)研究一种简单的基于非阵列的多重检测和测量MMPs/ADAMS活性的方法，以提高检测效率，降低传感器制造和检测成本；以及3)开发一种便携式固态纳米孔蛋白酶检测技术，用于潜在的移动实验室检测癌症生物标记物。预计这些研究将对各种重要的领域产生广泛的影响，如生物学、纳米技术、制药工业、一般毒理学和生物传感。教育计划的组成部分包括：将研究成果纳入课堂教学；从代表性不足的群体中积极招聘本科生和研究生；为高中生设立暑期学者研究方案；以及组织跨学科专题讨论会。这一跨学科项目为所有参与者提供了理想的培训机会，特别是在未来的职业机会方面。

Non-technical:This award by the Biomaterials Program in the Division of Materials Research to the Illinois Institute of Technology is to develop next-generation of biomaterials based on two groups of biocatalysts, namely MMPs (matrix metalloproteinases) and ADAMs (a disintegrin and metalloproteinases) for possible applications in sensing and other technologies. MMPs and ADAMs are enzymes that are responsible for extracellular matrix degradation and tissue remodeling, and play important roles in various biological, physiological, and pathological processes. They are currently under intensive investigation as novel biomarkers and potential therapeutic targets for the early detection and possible treatment of human cancers. However, the current MMP/ADAM assays suffer from low specificity and poor selectivity, which results in their limited utility for any applications. This project offers a new label-free strategy, which combines nanopore sensing and substrate-based proteinase assay, to accomplish highly selective and sensitive measurement of the activity of MMPs/ADAMs. The proposed studies are expected to lead to a better understanding of molecular and ionic transport, as well as to develop a versatile tool for various applications, including biosensing, studying covalent and non-covalent bonding interactions, investigating biomolecular folding and unfolding, and exploring enzyme kinetics. In addition to research, educational and outreach components are also an integral part of this project. The newest research findings from this work will be incorporated in classroom teaching for the benefit of students not directly involved in this research. The PI will recruit undergraduate and graduate students (especially Hispanic and African American minority groups) to participate in the research through established local programs. The PI will disseminate nanopore sensing technology by organizing interdisciplinary symposiums to promote idea exchange and facilitate national & international collaborations among peers and general public. Technical:This project will explore a new strategy to measure the activities of MMPs (matrix metalloproteinases) and ADAMs (a disintegrin and metalloproteinases), and this is accomplished by real-time monitoring of the ionic current modulations caused by the protease-substrate peptide interactions in the nanopore. By taking advantage of both the substrate specificity and the substrate cleavage sites, the nanopore sensor offers a potential to significantly improve the selectivity and accuracy of the MMPs/ADAMs assays. The three objectives of this project will be: 1) to investigate various factors which affect peptide transport in a nano-channel, and gain understanding of the underlying mechanism; 2) to study a simple non-array-based methodology for the multiplex detection and measurement of activities of MMPs/ADAMs to improve assay efficiency, and reduce the sensor manufacturing and assay cost; and 3) to develop a portable solid-state nanopore-based protease detection technique for potential mobile-lab detection of cancer biomarkers for point-of-care applications. The studies are expected to have broad impact on a variety of areas where analyses of proteases are important, such as biology, nanotechnology, pharmaceutical industry, toxicology in general, and biosensing. The components of the educational plan include: incorporation of research findings into classroom teaching; active recruitment of undergraduate and graduate students from underrepresented groups; establishment of a summer scholar research program for high school students; and organizing interdisciplinary symposiums. This interdisciplinary project offers an ideal training opportunity for all participants, particularly with respect to future career opportunities.