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

Illinois Institute Of Technology, Chicago IL

Investigators

Abstract

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.