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A Zebrafish Model of Metastatic Colonization Pinpoints Cellular Mechanisms of Circulating Tumor Cell Extravasation
Tyler A Allen1, Mark M Cullen1, Nathan Hawkey1
1Duke Cancer Institute, Duke University Medical Center, Durham, NC, United States.
Abstract:
Metastasis is a multistep process in which cells must detach, migrate/invade local structures, intravasate, circulate, extravasate, and colonize. A full understanding of the complexity of this process has been limited by the lack of ability to study these steps in isolation with detailed molecular analyses. Leveraging a comparative oncology approach, we injected canine osteosarcoma cells into the circulation of transgenic zebrafish with fluorescent blood vessels in a biologically dynamic metastasis extravasation model. Circulating tumor cell clusters that successfully extravasated the vasculature as multicellular units were isolated under intravital imaging (n = 6). These extravasation-positive tumor cell clusters sublines were then molecularly profiled by RNA-Seq. Using a systems-level analysis, we pinpointed the downregulation of KRAS signaling, immune pathways, and extracellular matrix (ECM) organization as enriched in extravasated cells (p < 0.05). Within the extracellular matrix remodeling pathway, we identified versican (VCAN) as consistently upregulated and central to the ECM gene regulatory network (p < 0.05). Versican expression is prognostic for a poorer metastasis-free and overall survival in patients with osteosarcoma. Together, our results provide a novel experimental framework to study discrete steps in the metastatic process. Using this system, we identify the versican/ECM network dysregulation as a potential contributor to osteosarcoma circulating tumor cell metastasis.
Insights
Researchers developed a novel zebrafish model to study cancer metastasis. They found that versican (VCAN) upregulation in extracellular matrix remodeling contributes to osteosarcoma cell extravasation and metastasis.
Area of Science:
- Comparative oncology
- Cancer metastasis research
- Zebrafish models
Background:
- Metastasis involves multiple complex steps: detachment, migration, invasion, intravasation, circulation, extravasation, and colonization.
- Studying these individual steps molecularly has been challenging.
- A comparative oncology approach using zebrafish offers a dynamic model for metastasis research.
Purpose of the Study:
- To develop a novel experimental framework for studying discrete steps of cancer metastasis.
- To molecularly profile circulating tumor cell clusters that successfully extravasate.
- To identify molecular contributors to osteosarcoma metastasis.
Main Methods:
- Injected canine osteosarcoma cells into transgenic zebrafish with fluorescent blood vessels.
- Utilized intravital imaging to isolate extravasation-positive tumor cell clusters.
- Performed RNA-sequencing and systems-level analysis on isolated cell clusters.
Main Results:
- Extravasated cells showed downregulation of KRAS signaling, immune pathways, and extracellular matrix (ECM) organization.
- Versican (VCAN) was identified as consistently upregulated within the ECM remodeling pathway.
- VCAN expression is linked to poorer metastasis-free and overall survival in osteosarcoma patients.
Conclusions:
- A novel zebrafish model enables the study of specific metastasis steps.
- Upregulation of versican (VCAN) and dysregulation of the ECM network may drive osteosarcoma metastasis.
- This study provides a new framework and identifies VCAN as a potential therapeutic target.
