High-throughput protease activity cytometry reveals dose-dependent heterogeneity in PMA-mediated ADAM17 activation
As key components of autocrine signaling, pericellular proteases, a disintegrin and metalloproteinases (ADAMs) in particular, are known to impact the microenvironment of individual cells and have significant implications in various pathological situations including cancer, inflammatory and vascular...
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| Format: | Article |
| Language: | en_US |
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Royal Society of Chemistry
2017
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| Online Access: | http://hdl.handle.net/1721.1/106950 https://orcid.org/0000-0003-3150-6170 https://orcid.org/0000-0003-1224-8153 https://orcid.org/0000-0001-7215-1439 |
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| author | Wu, Lidan Claas, Allison Mary Sarkar, Aniruddh Lauffenburger, Douglas A Han, Jongyoon |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Wu, Lidan Claas, Allison Mary Sarkar, Aniruddh Lauffenburger, Douglas A Han, Jongyoon |
| author_sort | Wu, Lidan |
| collection | MIT |
| description | As key components of autocrine signaling, pericellular proteases, a disintegrin and metalloproteinases (ADAMs) in particular, are known to impact the microenvironment of individual cells and have significant implications in various pathological situations including cancer, inflammatory and vascular diseases. There is great incentive to develop a high-throughput platform for single-cell measurement of pericellular protease activity, as it is essential for studying the heterogeneity of protease response and the corresponding cell behavioral consequences. In this work, we developed a microfluidic platform to simultaneously monitor protease activity of many single cells in a time-dependent manner. This platform isolates individual microwells rapidly on demand and thus allows single-cell activity measurement of both cell-surface and secreted proteases by confining individual cells with diffusive FRET-based substrates. With this platform, we observed dose-dependent heterogeneous protease activation of HepG2 cells treated with phorbol 12-myristate 13-acetate (PMA). To study the temporal behavior of PMA-induced protease response, we monitored the pericellular protease activity of the same single cells during three different time periods and revealed the diversity in the dynamic patterns of single-cell protease activity profile upon PMA stimulation. The unique temporal information of single-cell protease response can help unveil the complicated functional role of pericellular proteases. |
| first_indexed | 2024-09-23T16:46:28Z |
| format | Article |
| id | mit-1721.1/106950 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T16:46:28Z |
| publishDate | 2017 |
| publisher | Royal Society of Chemistry |
| record_format | dspace |
| spelling | mit-1721.1/1069502022-10-03T08:14:39Z High-throughput protease activity cytometry reveals dose-dependent heterogeneity in PMA-mediated ADAM17 activation Wu, Lidan Claas, Allison Mary Sarkar, Aniruddh Lauffenburger, Douglas A Han, Jongyoon Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science Massachusetts Institute of Technology. Research Laboratory of Electronics Singapore-MIT Alliance in Research and Technology (SMART) Wu, Lidan Claas, Allison Mary Sarkar, Aniruddh Lauffenburger, Douglas A Han, Jongyoon As key components of autocrine signaling, pericellular proteases, a disintegrin and metalloproteinases (ADAMs) in particular, are known to impact the microenvironment of individual cells and have significant implications in various pathological situations including cancer, inflammatory and vascular diseases. There is great incentive to develop a high-throughput platform for single-cell measurement of pericellular protease activity, as it is essential for studying the heterogeneity of protease response and the corresponding cell behavioral consequences. In this work, we developed a microfluidic platform to simultaneously monitor protease activity of many single cells in a time-dependent manner. This platform isolates individual microwells rapidly on demand and thus allows single-cell activity measurement of both cell-surface and secreted proteases by confining individual cells with diffusive FRET-based substrates. With this platform, we observed dose-dependent heterogeneous protease activation of HepG2 cells treated with phorbol 12-myristate 13-acetate (PMA). To study the temporal behavior of PMA-induced protease response, we monitored the pericellular protease activity of the same single cells during three different time periods and revealed the diversity in the dynamic patterns of single-cell protease activity profile upon PMA stimulation. The unique temporal information of single-cell protease response can help unveil the complicated functional role of pericellular proteases. National Institutes of Health (U.S.) (Grant R01-CA096504) Singapore-MIT Alliance for Research and Technology (SMART) 2017-02-15T21:48:21Z 2017-02-15T21:48:21Z 2015-03 2015-01 Article http://purl.org/eprint/type/JournalArticle 1757-9694 1757-9708 http://hdl.handle.net/1721.1/106950 Wu, Lidan et al. “High-Throughput Protease Activity Cytometry Reveals Dose-Dependent Heterogeneity in PMA-Mediated ADAM17 Activation.” Integr. Biol. 7.5 (2015): 513–524. https://orcid.org/0000-0003-3150-6170 https://orcid.org/0000-0003-1224-8153 https://orcid.org/0000-0001-7215-1439 en_US http://dx.doi.org/10.1039/c5ib00019j Integrative Biology Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf Royal Society of Chemistry PMC |
| spellingShingle | Wu, Lidan Claas, Allison Mary Sarkar, Aniruddh Lauffenburger, Douglas A Han, Jongyoon High-throughput protease activity cytometry reveals dose-dependent heterogeneity in PMA-mediated ADAM17 activation |
| title | High-throughput protease activity cytometry reveals dose-dependent heterogeneity in PMA-mediated ADAM17 activation |
| title_full | High-throughput protease activity cytometry reveals dose-dependent heterogeneity in PMA-mediated ADAM17 activation |
| title_fullStr | High-throughput protease activity cytometry reveals dose-dependent heterogeneity in PMA-mediated ADAM17 activation |
| title_full_unstemmed | High-throughput protease activity cytometry reveals dose-dependent heterogeneity in PMA-mediated ADAM17 activation |
| title_short | High-throughput protease activity cytometry reveals dose-dependent heterogeneity in PMA-mediated ADAM17 activation |
| title_sort | high throughput protease activity cytometry reveals dose dependent heterogeneity in pma mediated adam17 activation |
| url | http://hdl.handle.net/1721.1/106950 https://orcid.org/0000-0003-3150-6170 https://orcid.org/0000-0003-1224-8153 https://orcid.org/0000-0001-7215-1439 |
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