A High-Throughput Assay for Collagen Secretion Suggests an Unanticipated Role for Hsp90 in Collagen Production
Collagen overproduction is a feature of fibrosis and cancer, while insufficient deposition of functional collagen molecules and/or the secretion of malformed collagen is common in genetic disorders like osteogenesis imperfecta. Collagen secretion is an appealing therapeutic target in these and other...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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American Chemical Society (ACS)
2020
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| Online Access: | https://hdl.handle.net/1721.1/125610 |
| _version_ | 1811070162677268480 |
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| author | Wong, Madeline Y. Doan, Ngoc Duc DiChiara, Andrew Stephen Papa, Louis John Cheah, Jaime H Soule, Christian K. Watson, Nicki E. Shoulders, Matthew D. |
| author2 | Massachusetts Institute of Technology. Department of Chemistry |
| author_facet | Massachusetts Institute of Technology. Department of Chemistry Wong, Madeline Y. Doan, Ngoc Duc DiChiara, Andrew Stephen Papa, Louis John Cheah, Jaime H Soule, Christian K. Watson, Nicki E. Shoulders, Matthew D. |
| author_sort | Wong, Madeline Y. |
| collection | MIT |
| description | Collagen overproduction is a feature of fibrosis and cancer, while insufficient deposition of functional collagen molecules and/or the secretion of malformed collagen is common in genetic disorders like osteogenesis imperfecta. Collagen secretion is an appealing therapeutic target in these and other diseases, as secretion directly connects intracellular biosynthesis to collagen deposition and biological function in the extracellular matrix. However, small molecule and biological methods to tune collagen secretion are severely lacking. Their discovery could prove useful not only in the treatment of disease, but also in providing tools for better elucidating mechanisms of collagen biosynthesis. We developed a cell-based, high-throughput luminescent assay of collagen type I secretion and used it to screen for small molecules that selectively enhance or inhibit that process. Among several validated hits, the Hsp90 inhibitor 17-allylaminogeldanamycin (17-AAG) robustly decreases the secretion of collagen-I by our model cell line and by human primary cells. In these systems, 17-AAG and other pan-isoform Hsp90 inhibitors reduce collagen-I secretion post-translationally and are not global inhibitors of protein secretion. Surprisingly, the consequences of Hsp90 inhibitors cannot be attributed to inhibition of the endoplasmic reticulum's Hsp90 isoform, Grp94. Instead, collagen-I secretion likely depends on the activity of cytosolic Hsp90 chaperones, even though such chaperones cannot directly engage nascent collagen molecules. Our results highlight the value of a cell-based high-throughput screen for selective modulators of collagen secretion and suggest an unanticipated role for cytosolic Hsp90 in collagen secretion. |
| first_indexed | 2024-09-23T08:29:00Z |
| format | Article |
| id | mit-1721.1/125610 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T08:29:00Z |
| publishDate | 2020 |
| publisher | American Chemical Society (ACS) |
| record_format | dspace |
| spelling | mit-1721.1/1256102022-09-30T09:23:55Z A High-Throughput Assay for Collagen Secretion Suggests an Unanticipated Role for Hsp90 in Collagen Production Wong, Madeline Y. Doan, Ngoc Duc DiChiara, Andrew Stephen Papa, Louis John Cheah, Jaime H Soule, Christian K. Watson, Nicki E. Shoulders, Matthew D. Massachusetts Institute of Technology. Department of Chemistry Whitehead Institute for Biomedical Research Koch Institute for Integrative Cancer Research at MIT Collagen overproduction is a feature of fibrosis and cancer, while insufficient deposition of functional collagen molecules and/or the secretion of malformed collagen is common in genetic disorders like osteogenesis imperfecta. Collagen secretion is an appealing therapeutic target in these and other diseases, as secretion directly connects intracellular biosynthesis to collagen deposition and biological function in the extracellular matrix. However, small molecule and biological methods to tune collagen secretion are severely lacking. Their discovery could prove useful not only in the treatment of disease, but also in providing tools for better elucidating mechanisms of collagen biosynthesis. We developed a cell-based, high-throughput luminescent assay of collagen type I secretion and used it to screen for small molecules that selectively enhance or inhibit that process. Among several validated hits, the Hsp90 inhibitor 17-allylaminogeldanamycin (17-AAG) robustly decreases the secretion of collagen-I by our model cell line and by human primary cells. In these systems, 17-AAG and other pan-isoform Hsp90 inhibitors reduce collagen-I secretion post-translationally and are not global inhibitors of protein secretion. Surprisingly, the consequences of Hsp90 inhibitors cannot be attributed to inhibition of the endoplasmic reticulum's Hsp90 isoform, Grp94. Instead, collagen-I secretion likely depends on the activity of cytosolic Hsp90 chaperones, even though such chaperones cannot directly engage nascent collagen molecules. Our results highlight the value of a cell-based high-throughput screen for selective modulators of collagen secretion and suggest an unanticipated role for cytosolic Hsp90 in collagen secretion. National Institutes of Health (U.S.) (Grant 1R03AR067503) National Institutes of Health (U.S.) (Grant 1R01AR071443) National Institutes of Health (U.S.). Ruth Kirschstein Pre-Doctoral Fellowship (1F31AR067615) National Cancer Institute (U.S.) (Cancer Center Support (core) Grant P30-CA14051) 2020-06-02T16:47:48Z 2020-06-02T16:47:48Z 2018-05 2020-01-08T17:02:47Z Article http://purl.org/eprint/type/JournalArticle 0006-2960 https://hdl.handle.net/1721.1/125610 Wong, Madeline Y. et al. “A High-Throughput Assay for Collagen Secretion Suggests an Unanticipated Role for Hsp90 in Collagen Production.” Biochemistry 57 (2018): 2814-2827 © 2018 The Author(s) en https://dx.doi.org/10.1021/ACS.BIOCHEM.8B00378 Biochemistry Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf American Chemical Society (ACS) PMC |
| spellingShingle | Wong, Madeline Y. Doan, Ngoc Duc DiChiara, Andrew Stephen Papa, Louis John Cheah, Jaime H Soule, Christian K. Watson, Nicki E. Shoulders, Matthew D. A High-Throughput Assay for Collagen Secretion Suggests an Unanticipated Role for Hsp90 in Collagen Production |
| title | A High-Throughput Assay for Collagen Secretion Suggests an Unanticipated Role for Hsp90 in Collagen Production |
| title_full | A High-Throughput Assay for Collagen Secretion Suggests an Unanticipated Role for Hsp90 in Collagen Production |
| title_fullStr | A High-Throughput Assay for Collagen Secretion Suggests an Unanticipated Role for Hsp90 in Collagen Production |
| title_full_unstemmed | A High-Throughput Assay for Collagen Secretion Suggests an Unanticipated Role for Hsp90 in Collagen Production |
| title_short | A High-Throughput Assay for Collagen Secretion Suggests an Unanticipated Role for Hsp90 in Collagen Production |
| title_sort | high throughput assay for collagen secretion suggests an unanticipated role for hsp90 in collagen production |
| url | https://hdl.handle.net/1721.1/125610 |
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