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

非技术性：伊利诺伊理工学院材料研究部生物材料项目的这一奖项旨在开发基于两组生物催化剂的下一代生物材料，即MMP（基质金属蛋白酶）和ADAM（去整合素和金属蛋白酶），用于传感和其他技术的可能应用。MMPs和ADAMs是负责细胞外基质降解和组织重塑的酶，并且在各种生物、生理和病理过程中发挥重要作用。它们目前正在作为新的生物标志物和潜在的治疗靶点进行深入研究，用于人类癌症的早期检测和可能的治疗。然而，目前的MMP/ADAM测定具有低特异性和差选择性，这导致其对于任何应用的有限效用。该项目提供了一种新的无标记策略，该策略结合了纳米孔传感和基于底物的蛋白酶测定，以实现MMPs/ADAMs活性的高选择性和灵敏度测量。拟议的研究预计将导致更好地了解分子和离子的运输，以及开发一个多功能的工具，用于各种应用，包括生物传感，研究共价和非共价键的相互作用，调查生物分子的折叠和展开，并探索酶动力学。除研究外，教育和外联部分也是该项目的一个组成部分。 这项工作的最新研究成果将被纳入课堂教学中，以使没有直接参与这项研究的学生受益。PI将招募本科生和研究生（特别是西班牙裔和非洲裔美国少数民族）通过既定的当地计划参与研究。PI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，并促进同行和公众之间的国家国际合作。&技术：该项目将探索一种新的策略来测量MMP（基质金属蛋白酶）和ADAM（一种去整合素和金属蛋白酶）的活性，这是通过实时监测纳米孔中蛋白酶-底物肽相互作用引起的离子电流调制来实现的。通过利用底物特异性和底物切割位点，纳米孔传感器提供了显著提高MMP/ADAM测定的选择性和准确性的潜力。本项目的三个目标是：1）研究影响多肽在纳米通道中转运的各种因素，并了解其内在机制; 2）研究一种简单的非阵列化方法，用于MMPs/ADAMs活性的多重检测，以提高检测效率，降低传感器的制造和检测成本;和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志物，以用于护理点应用。预计这些研究将对蛋白酶分析非常重要的各种领域产生广泛影响，例如生物学，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;积极招收代表性不足群体的本科生和研究生;为高中生设立暑期学者研究方案;组织跨学科研讨会。这个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.