Activatable zymography probes enable in situ localization of protease dysregulation in cancer
©2020 American Association for Cancer Research. Recent years have seen the emergence of conditionally activated diagnostics and therapeutics that leverage protease-cleavable peptide linkers to enhance their specificity for cancer. However, due to a lack of methods to measure and localize protease ac...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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American Association for Cancer Research (AACR)
2021
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| Online Access: | https://hdl.handle.net/1721.1/134556 |
| _version_ | 1811006732252479488 |
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| author | Soleimany, Ava P Kirkpatrick, Jesse D Su, Susan Dudani, Jaideep S Zhong, Qian Bekdemir, Ahmet Bhatia, Sangeeta N |
| author_facet | Soleimany, Ava P Kirkpatrick, Jesse D Su, Susan Dudani, Jaideep S Zhong, Qian Bekdemir, Ahmet Bhatia, Sangeeta N |
| author_sort | Soleimany, Ava P |
| collection | MIT |
| description | ©2020 American Association for Cancer Research. Recent years have seen the emergence of conditionally activated diagnostics and therapeutics that leverage protease-cleavable peptide linkers to enhance their specificity for cancer. However, due to a lack of methods to measure and localize protease activity directly within the tissue microenvironment, the design of protease-activated agents has been necessarily empirical, yielding suboptimal results when translated to patients. To address the need for spatially resolved protease activity profiling in cancer, we developed a new class of in situ probes that can be applied to fresh-frozen tissue sections in a manner analogous to immunofluorescence staining. These activatable zymography probes (AZP) detected dysregulated protease activity in human prostate cancer biopsy samples, enabling disease classification. AZPs were leveraged within a generalizable framework to design conditional cancer diagnostics and therapeutics and showcased in the Hi-Myc mouse model of prostate cancer, which models features of early pathogenesis. Multiplexed screening against barcoded substrates yielded a peptide, S16, that was robustly and specifically cleaved by tumor-associated metalloproteinases in the Hi-Myc model. In situ labeling with an AZP incorporating S16 revealed a potential role of metalloproteinase dysregulation in proliferative, premalignant Hi-Myc prostatic glands. Systemic administration of an in vivo imaging probe incorporating S16 perfectly classified diseased and healthy prostates, supporting the relevance of ex vivo activity assays to in vivo translation. We envision AZPs will enable new insights into the biology of protease dysregulation in cancer and accelerate the development of conditional diagnostics and therapeutics for multiple cancer types. Significance: Visualization of protease activity within the native tissue context using AZPs provides new biological insights into protease dysregulation in cancer and guides the design of conditional diagnostics and therapeutics. |
| first_indexed | 2024-09-23T16:56:47Z |
| format | Article |
| id | mit-1721.1/134556 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T16:56:47Z |
| publishDate | 2021 |
| publisher | American Association for Cancer Research (AACR) |
| record_format | dspace |
| spelling | mit-1721.1/1345562021-10-28T03:04:41Z Activatable zymography probes enable in situ localization of protease dysregulation in cancer Soleimany, Ava P Kirkpatrick, Jesse D Su, Susan Dudani, Jaideep S Zhong, Qian Bekdemir, Ahmet Bhatia, Sangeeta N ©2020 American Association for Cancer Research. Recent years have seen the emergence of conditionally activated diagnostics and therapeutics that leverage protease-cleavable peptide linkers to enhance their specificity for cancer. However, due to a lack of methods to measure and localize protease activity directly within the tissue microenvironment, the design of protease-activated agents has been necessarily empirical, yielding suboptimal results when translated to patients. To address the need for spatially resolved protease activity profiling in cancer, we developed a new class of in situ probes that can be applied to fresh-frozen tissue sections in a manner analogous to immunofluorescence staining. These activatable zymography probes (AZP) detected dysregulated protease activity in human prostate cancer biopsy samples, enabling disease classification. AZPs were leveraged within a generalizable framework to design conditional cancer diagnostics and therapeutics and showcased in the Hi-Myc mouse model of prostate cancer, which models features of early pathogenesis. Multiplexed screening against barcoded substrates yielded a peptide, S16, that was robustly and specifically cleaved by tumor-associated metalloproteinases in the Hi-Myc model. In situ labeling with an AZP incorporating S16 revealed a potential role of metalloproteinase dysregulation in proliferative, premalignant Hi-Myc prostatic glands. Systemic administration of an in vivo imaging probe incorporating S16 perfectly classified diseased and healthy prostates, supporting the relevance of ex vivo activity assays to in vivo translation. We envision AZPs will enable new insights into the biology of protease dysregulation in cancer and accelerate the development of conditional diagnostics and therapeutics for multiple cancer types. Significance: Visualization of protease activity within the native tissue context using AZPs provides new biological insights into protease dysregulation in cancer and guides the design of conditional diagnostics and therapeutics. 2021-10-27T20:05:34Z 2021-10-27T20:05:34Z 2020 2020-12-02T18:46:57Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/134556 en 10.1158/0008-5472.can-20-2410 Cancer Research Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf American Association for Cancer Research (AACR) bioRxiv |
| spellingShingle | Soleimany, Ava P Kirkpatrick, Jesse D Su, Susan Dudani, Jaideep S Zhong, Qian Bekdemir, Ahmet Bhatia, Sangeeta N Activatable zymography probes enable in situ localization of protease dysregulation in cancer |
| title | Activatable zymography probes enable in situ localization of protease dysregulation in cancer |
| title_full | Activatable zymography probes enable in situ localization of protease dysregulation in cancer |
| title_fullStr | Activatable zymography probes enable in situ localization of protease dysregulation in cancer |
| title_full_unstemmed | Activatable zymography probes enable in situ localization of protease dysregulation in cancer |
| title_short | Activatable zymography probes enable in situ localization of protease dysregulation in cancer |
| title_sort | activatable zymography probes enable in situ localization of protease dysregulation in cancer |
| url | https://hdl.handle.net/1721.1/134556 |
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