FRET Sensor-Modified Synthetic Hydrogels for Real-Time Monitoring of Cell-Derived Matrix Metalloproteinase Activity using Fluorescence Lifetime Imaging

Ziqian Yan; Thomas Kavanagh; Ricardo da Silva Harrabi; Suzie Lust; Chunling Tang; Rebecca Beavil; Manuel M. Mueller; Andrew Beavil; Simon Ameer-Beg; Ricardo M. P. da Silva; Eileen Gentleman

doi:10.1002/adfm.202309711

FRET Sensor-Modified Synthetic Hydrogels for Real-Time Monitoring of Cell-Derived Matrix Metalloproteinase Activity using Fluorescence Lifetime Imaging

Ziqian Yan, Thomas Kavanagh, Ricardo da Silva Harrabi, Suzie Lust, Chunling Tang, Rebecca Beavil, Manuel M. Mueller, Andrew Beavil, Simon Ameer-Beg, Ricardo M. P. da Silva, Eileen Gentleman

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Abstract

Matrix 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.

Original language	English
Article number	2309711
Journal	Advanced Functional Materials
Volume	34
Issue number	21
DOIs	https://doi.org/10.1002/adfm.202309711
Publication status	Published - 22 May 2024

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Yan et al MMP FRET sensor Final Author AcceptedAccepted author manuscript, 3.62 MBLicence: CC BY

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Yan, Z., Kavanagh, T., da Silva Harrabi, R., Lust, S., Tang, C., Beavil, R., Mueller, M. M., Beavil, A., Ameer-Beg, S., M. P. da Silva, R., & Gentleman, E. (2024). FRET Sensor-Modified Synthetic Hydrogels for Real-Time Monitoring of Cell-Derived Matrix Metalloproteinase Activity using Fluorescence Lifetime Imaging. Advanced Functional Materials, 34(21), Article 2309711. https://doi.org/10.1002/adfm.202309711

@article{798bf9107d204746b7c6aeb1410586cd,

title = "FRET Sensor-Modified Synthetic Hydrogels for Real-Time Monitoring of Cell-Derived Matrix Metalloproteinase Activity using Fluorescence Lifetime Imaging",

abstract = "Matrix 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 F{\"o}rster 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.",

author = "Ziqian Yan and Thomas Kavanagh and {da Silva Harrabi}, Ricardo and Suzie Lust and Chunling Tang and Rebecca Beavil and Mueller, {Manuel M.} and Andrew Beavil and Simon Ameer-Beg and {M. P. da Silva}, Ricardo and Eileen Gentleman",

note = "Funding Information: Z.Y. acknowledges a King's Postgraduate International Scholarship from King's College London. S.T.L. acknowledges the UK Medical Research Council (MR/N013700/1) for funding through the MRC Doctoral Training Partnership in Biomedical Sciences at King's College London. R.M.P.d.S. acknowledges a King's Prize fellowship supported by the Wellcome Trust (Institutional Strategic Support Fund), King's College London, and the London Law Trust. M.M. acknowledges funding from the Wellcome Trust and the Royal Society for a Sir Henry Wellcome Fellowship (202250/Z/16/Z). E.G. acknowledges funding from the UK Engineering and Physical Sciences Council (EP/V04723X/1) and the Rosetrees Trust. The authors are extremely grateful to Dr. Sashko Damjanovski at the University of Western Ontario for providing the MCF7 MMP14 cell line. Publisher Copyright: {\textcopyright} 2024 The Authors. Advanced Functional Materials published by Wiley-VCH GmbH.",

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doi = "10.1002/adfm.202309711",

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Yan, Z , Kavanagh, T, da Silva Harrabi, R, Lust, S , Tang, C , Beavil, R , Mueller, MM , Beavil, A , Ameer-Beg, S , M. P. da Silva, R & Gentleman, E 2024, 'FRET Sensor-Modified Synthetic Hydrogels for Real-Time Monitoring of Cell-Derived Matrix Metalloproteinase Activity using Fluorescence Lifetime Imaging', Advanced Functional Materials, vol. 34, no. 21, 2309711. https://doi.org/10.1002/adfm.202309711

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T1 - FRET Sensor-Modified Synthetic Hydrogels for Real-Time Monitoring of Cell-Derived Matrix Metalloproteinase Activity using Fluorescence Lifetime Imaging

AU - Yan, Ziqian

AU - Kavanagh, Thomas

AU - da Silva Harrabi, Ricardo

AU - Lust, Suzie

AU - Tang, Chunling

AU - Beavil, Rebecca

AU - Mueller, Manuel M.

AU - Beavil, Andrew

AU - Ameer-Beg, Simon

AU - M. P. da Silva, Ricardo

AU - Gentleman, Eileen

N1 - Funding Information: Z.Y. acknowledges a King's Postgraduate International Scholarship from King's College London. S.T.L. acknowledges the UK Medical Research Council (MR/N013700/1) for funding through the MRC Doctoral Training Partnership in Biomedical Sciences at King's College London. R.M.P.d.S. acknowledges a King's Prize fellowship supported by the Wellcome Trust (Institutional Strategic Support Fund), King's College London, and the London Law Trust. M.M. acknowledges funding from the Wellcome Trust and the Royal Society for a Sir Henry Wellcome Fellowship (202250/Z/16/Z). E.G. acknowledges funding from the UK Engineering and Physical Sciences Council (EP/V04723X/1) and the Rosetrees Trust. The authors are extremely grateful to Dr. Sashko Damjanovski at the University of Western Ontario for providing the MCF7 MMP14 cell line. Publisher Copyright: © 2024 The Authors. Advanced Functional Materials published by Wiley-VCH GmbH.

PY - 2024/5/22

Y1 - 2024/5/22

N2 - Matrix 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.

AB - Matrix 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.

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U2 - 10.1002/adfm.202309711

DO - 10.1002/adfm.202309711

M3 - Article

SN - 1616-301X

VL - 34

JO - Advanced Functional Materials

JF - Advanced Functional Materials

IS - 21

M1 - 2309711

ER -

FRET Sensor-Modified Synthetic Hydrogels for Real-Time Monitoring of Cell-Derived Matrix Metalloproteinase Activity using Fluorescence Lifetime Imaging

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