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Thesis Format

Integrated Article

Degree

Doctor of Philosophy

Program

Pathology and Laboratory Medicine

Supervisor

Khan, Zia A.

2nd Supervisor

Chakraborty, Chandan

Co-Supervisor

Abstract

Cancer metastasis is a multistep process that begins with the invasion of tumour cells into the stroma and migration towards the blood vessels. Tumour cells that have entered the bloodstream must then survive and leave by a process known as extravasation. Finally, extravasated cells proliferate and establish the secondary site in the metastatic cascade. Although extravasation encompasses key events during cancer cell invasion to aid in the development of effective treatments, an in vivo model that rapidly, reproducibly and economically recapitulates cancer cell extravasation is needed. Therefore, the objectives of my research were to 1) establish and validate an in vivo model of cancer cell extravasation, and 2) identify novel cellular and molecular events.

I used the chorioallantoic membrane of chicken embryos as a model system of extravasation as it provides an accessible and highly vascularized structure. The combination of the chorioallantoic membrane of chicken embryos, nanoscale flow cytometry, and confocal microscopy-based intravital imaging allowed me to observe that extravasating prostate cancer cells exhibited significant cell volume reduction. This reduction is suggestive of an invasive cell phenotype. However, cell volume reduction at certain threshold also decreased cancer cell extravasation efficiency. I also found that cancer cell released extravascular vesicles during extravasation, and an increase in extracellular vesicle release reduced cell volume. I then tested the hypothesis that extracellular vesicle release and extravasation may be linked to modes of cell death. Real-time imaging of extravasating cancer cells that released extracellular vesicles did not show activation of caspase-3. Activation of necroptosis, however, increased extracellular vesicle release and decreased cell extravasation and secondary colony formation. These results suggest that necroptosis may be targeted to induce extracellular vesicle release, decrease extravasation, and halt cancer metastasis.

Collectively, my work lays out the protocols for the use of the chorioallantoic membrane of chicken embryos as a model system to investigate cancer cell extravasation and invasion. Use of this model system allowed me to identify extracellular vesicle release during extravasation and discover that necroptosis may be a potential regulator of cancer metastasis.

Summary for Lay Audience

Cancer spreading to different areas in the body is known as metastasis and is the main cause of cancer-related deaths. Many underlying events in cancer metastasis are still poorly understood. To provide new insights, I first established a new model to study metastasis. I injected human prostate cancer cells into the blood vessel of chicken embryo lungs. I then monitored the cancer cells in real-time using microscopy. This technique was optimized, and results were published to allow other researchers to study metastasis in this quick, economical, and reliable model. For the second part of my part, I investigated the changes that take place in cancer cells when the cells move from the injected area to different areas. I observed that cancer cells shrink their sizes when they move out of the bloodstream to enter different sites. My results also indicate that this shrinking of cancer cells may have been made possible by a release of small portions of the cell. I reasoned that if this release can be artificially increased, it would possibly kill more cancer cells and provide a new target to stop metastasis. Indeed, my results show that increasing the release of cell portions reduces the ability of cancer cells to move to secondary sites. Therefore, my studies lay the foundation for a new therapeutic target for cancer patients.

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

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