Intravital imaging tumor screen used to identify novel metastasis-blocking therapeutic targets
Authors:Konstantin Stoletov1, Lian Willetts1, Perrin H. Beatty1 and John D. Lewis1
1 Department of Oncology, University of Alberta, Edmonton, Alberta, Canada, T6G 2E1.
Keywords:
ex ovo chick embryo model, fluorescence time-lapse intravital imaging, high throughput sequencing, intravasation, metastatic cancer, shRNA library, therapeutic targets.
Corresponding Author(s):
Conflict of interest statement:
The authors declare no competing interests.
Please cite this article as:
Konstantin Stoletov, Lian Willetts, Perrin H. Beatty and John D. Lewis (2018). Intravital imaging tumor screen used to identify novel metastasis-blocking therapeutic targets. Cell Stress 2(10): 275-278. doi: 10.15698/cst2018.10.159
© 2018 Stoletov et al. This is an open-access article released under the terms of the Creative Commons Attribution (CC BY) license, which allows the unrestricted use, distribution, and reproduction in any medium, provided the original author and source are acknowledged.
Abstract:
Cancer cell motility is a key driver of metastasis. Although the intravasation of cancer cells into the blood stream is highly dependent on their motility and metastatic dissemination is the primary cause of cancer related deaths, current therapeutic strategies do not target the genes and proteins that are essential for cell motility. A primary reason for this is because the identification of cell motility-related genes that are relevant in vivo requires the visualization of metastatic lesions forming in an appropriate in vivo model. The cancer research community has lacked an in vivo and intravital metastatic cancer model that could be imaged as motility developed, in real-time. To address this, we developed a novel quantitative in vivo screening platform based on intravital imaging in shell-less ex ovo chick embryos. We applied this imaging approach to screen a human genome-wide short hairpin RNA library (shRNA) versus the highly motile head and neck cancer cells (HEp3 cell line) introduced into the chorioallantoic membrane (CAM) of chick embryos and identified multiple novel in vivo cancer cell motility-associated genes. When the expression of several of the identified genes was inhibited in the HEp3 tumors, we observed a nearly total block of spontaneous cancer metastasis.