Affiliation:
1. Centre for Craniofacial and Regenerative Biology King's College London London SE1 9RT UK
2. Richard Dimbleby Laboratory of Cancer Research School of Cancer and Pharmaceutical Sciences King's College London London SE1 1UL UK
3. Randall Centre of Cell and Molecular Biophysics King's College London London SE1 1UL UK
4. Department of Chemistry King's College London London SE1 1DB UK
5. Department of Biomedical Sciences University of Lausanne Lausanne 1005 Switzerland
Abstract
AbstractMatrix remodeling plays central roles in a range of physiological and pathological processes and is driven predominantly by the activity of matrix metalloproteinases (MMPs), which degrade extracellular matrix (ECM) proteins. How MMPs regulate cell and tissue dynamics is not well understood as in vivo approaches are lacking and many in vitro strategies cannot provide high‐resolution, quantitative measures of enzyme activity in situ within tissue‐like 3D microenvironments. Here, a Förster resonance energy transfer (FRET) sensor of MMP activity is incorporated into fully synthetic hydrogels that mimic many properties of the native ECM. Fluorescence lifetime imaging is then used to provide a real‐time, fluorophore concentration‐independent quantification of MMP activity, establishing a highly accurate, readily adaptable platform for studying MMP dynamics in situ. MCF7 human breast cancer cells encapsulated within hydrogels are then used to detect MMP activity both locally, at the sub‐micron level, and within the bulk hydrogel. This versatile platform may find use in a range of biological studies to explore questions in the dynamics of cancer metastasis, development, and tissue repair by providing high‐resolution, quantitative, and in situ readouts of local MMP activity within native tissue‐like environments.
Funder
Medical Research Council
Engineering and Physical Sciences Research Council
Rosetrees Trust
Subject
Electrochemistry,Condensed Matter Physics,Biomaterials,Electronic, Optical and Magnetic Materials