A Quantitative System for Studying Metastasis Using Transparent Zebrafish
Metastasis is the defining feature of advanced malignancy, yet remains challenging to study in laboratory environments. Here, we describe a high-throughput zebrafish system for comprehensive, in vivo assessment of metastatic biology. First, we generated several stable cell lines from melanomas of tr...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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American Association for Cancer Research
2017
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| Online Access: | http://hdl.handle.net/1721.1/107192 https://orcid.org/0000-0001-7603-8396 |
| _version_ | 1826213342875746304 |
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| author | Heilmann, S. Ratnakumar, K. Langdon, E. M. Kansler, E. R. Kim, I. S. Campbell, N. R. Perry, E. B. Kaufman, C. K. van Rooijen, E. Lee, W. Iacobuzio-Donahue, C. A. Zon, L. I. Xavier, J. B. White, R. M. Hynes, Richard O. McMahon, Amy J. |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Heilmann, S. Ratnakumar, K. Langdon, E. M. Kansler, E. R. Kim, I. S. Campbell, N. R. Perry, E. B. Kaufman, C. K. van Rooijen, E. Lee, W. Iacobuzio-Donahue, C. A. Zon, L. I. Xavier, J. B. White, R. M. Hynes, Richard O. McMahon, Amy J. |
| author_sort | Heilmann, S. |
| collection | MIT |
| description | Metastasis is the defining feature of advanced malignancy, yet remains challenging to study in laboratory environments. Here, we describe a high-throughput zebrafish system for comprehensive, in vivo assessment of metastatic biology. First, we generated several stable cell lines from melanomas of transgenic mitfa-BRAF[superscript V600E];p53[superscript −/−] fish. We then transplanted the melanoma cells into the transparent casper strain to enable highly quantitative measurement of the metastatic process at single-cell resolution. Using computational image analysis of the resulting metastases, we generated a metastasis score, μ, that can be applied to quantitative comparison of metastatic capacity between experimental conditions. Furthermore, image analysis also provided estimates of the frequency of metastasis-initiating cells (∼1/120,000 cells). Finally, we determined that the degree of pigmentation is a key feature defining cells with metastatic capability. The small size and rapid generation of progeny combined with superior imaging tools make zebrafish ideal for unbiased high-throughput investigations of cell-intrinsic or microenvironmental modifiers of metastasis. The approaches described here are readily applicable to other tumor types and thus serve to complement studies also employing murine and human cell culture systems. |
| first_indexed | 2024-09-23T15:47:33Z |
| format | Article |
| id | mit-1721.1/107192 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T15:47:33Z |
| publishDate | 2017 |
| publisher | American Association for Cancer Research |
| record_format | dspace |
| spelling | mit-1721.1/1071922022-09-29T16:11:46Z A Quantitative System for Studying Metastasis Using Transparent Zebrafish Heilmann, S. Ratnakumar, K. Langdon, E. M. Kansler, E. R. Kim, I. S. Campbell, N. R. Perry, E. B. Kaufman, C. K. van Rooijen, E. Lee, W. Iacobuzio-Donahue, C. A. Zon, L. I. Xavier, J. B. White, R. M. Hynes, Richard O. McMahon, Amy J. Massachusetts Institute of Technology. Department of Biology Koch Institute for Integrative Cancer Research at MIT Hynes, Richard O. McMahon, Amy J. Metastasis is the defining feature of advanced malignancy, yet remains challenging to study in laboratory environments. Here, we describe a high-throughput zebrafish system for comprehensive, in vivo assessment of metastatic biology. First, we generated several stable cell lines from melanomas of transgenic mitfa-BRAF[superscript V600E];p53[superscript −/−] fish. We then transplanted the melanoma cells into the transparent casper strain to enable highly quantitative measurement of the metastatic process at single-cell resolution. Using computational image analysis of the resulting metastases, we generated a metastasis score, μ, that can be applied to quantitative comparison of metastatic capacity between experimental conditions. Furthermore, image analysis also provided estimates of the frequency of metastasis-initiating cells (∼1/120,000 cells). Finally, we determined that the degree of pigmentation is a key feature defining cells with metastatic capability. The small size and rapid generation of progeny combined with superior imaging tools make zebrafish ideal for unbiased high-throughput investigations of cell-intrinsic or microenvironmental modifiers of metastasis. The approaches described here are readily applicable to other tumor types and thus serve to complement studies also employing murine and human cell culture systems. National Institutes of Health (U.S.) (Directors New Innovator Award DP2CA186572 and K08AR055368) Melanoma Research Alliance (Young Investigator Award) American Association for Cancer Research/American Society of Clinical Oncology (Young Investigator Award) Memorial Sloan-Kettering Cancer Center. Alan and Sandra Gerry Metastasis Research Initiative Howard Hughes Medical Institute 2017-03-06T16:47:27Z 2017-03-06T16:47:27Z 2015-08 2015-05 Article http://purl.org/eprint/type/JournalArticle 0008-5472 1538-7445 http://hdl.handle.net/1721.1/107192 Heilmann, S. et al. “A Quantitative System for Studying Metastasis Using Transparent Zebrafish.” Cancer Research 75.20 (2015): 4272–4282. https://orcid.org/0000-0001-7603-8396 en_US http://dx.doi.org/10.1158/0008-5472.can-14-3319 Cancer Research Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf American Association for Cancer Research PMC |
| spellingShingle | Heilmann, S. Ratnakumar, K. Langdon, E. M. Kansler, E. R. Kim, I. S. Campbell, N. R. Perry, E. B. Kaufman, C. K. van Rooijen, E. Lee, W. Iacobuzio-Donahue, C. A. Zon, L. I. Xavier, J. B. White, R. M. Hynes, Richard O. McMahon, Amy J. A Quantitative System for Studying Metastasis Using Transparent Zebrafish |
| title | A Quantitative System for Studying Metastasis Using Transparent Zebrafish |
| title_full | A Quantitative System for Studying Metastasis Using Transparent Zebrafish |
| title_fullStr | A Quantitative System for Studying Metastasis Using Transparent Zebrafish |
| title_full_unstemmed | A Quantitative System for Studying Metastasis Using Transparent Zebrafish |
| title_short | A Quantitative System for Studying Metastasis Using Transparent Zebrafish |
| title_sort | quantitative system for studying metastasis using transparent zebrafish |
| url | http://hdl.handle.net/1721.1/107192 https://orcid.org/0000-0001-7603-8396 |
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