About the Project
Taxol (paclitaxel) is a major drug used in the treatment of solid cancers (breast, lung, ovarian, bladder). However, resistance to taxol arises regularly and thus it is critical to find ways to circumvent this resistance. Our lab works on a family of proteins, Kinesin-13, that can depolymerize taxol-stabilized microtubules and are overexpressed in certain cancers resistant to taxol (Ganguly et al., 2011). We want to understand the mechanism that causes resistance to taxol and develop assays to identify taxol-resistant cancers that would become sensitive to the presence of Kinesin-13 microtubule depolymerases. This could then be used to design drugs that may inhibit the activity of Kinesin-13, and other microtubule depolymerizing drugs, based on our structural knowledge.
We have generated multiple cell lines that are sensitive and resistant to taxol. Upon acquisition of taxol resistance, the cell shape becomes more elongated. The project will involve inserting fluorescent markers (H2B, tubulin) stably into these cell lines using CRISPR technology. The student will then test whether the taxol-resistant cell lines become dependent on microtubule depolymerases using conditional knockouts. The student will use live-cell imaging and advanced quantitative microscopy to examine the dynamics of cell division and cytoskeletal organization throughout the cell cycle. They will develop machine-learning approaches to characterize the cell cycle behaviour and cytoskeleton changes linked to cell morphology. Using automated image analysis, the student will be able to develop algorithms to rapidly analyse which drugs are the most efficient at killing taxol-resistant cells. Other pathways and candidates that change during the acquisition of resistance to taxol will also be studied and evaluated as future drug targets. These data will indicate which pathways can be targeted in taxol-resistant cells and give leads for small molecule inhibitor screens, then performed at the IGMM or with CRUK drug discovery unit. This project will contribute to working towards personal medicine and personalized treatments following resistance to cancer.
References
Akbani, R., Becker, K.F., Carragher, N., Goldstein, T., de Koning, L., Korf, U., Liotta, L., Mills, G.B., Nishizuka, S.S., Pawlak, M., et al. (2014). Realizing the promise of reverse phase protein arrays for clinical, translational, and basic research: a workshop report: the RPPA (Reverse Phase Protein Array) society. Mol Cell Proteomics 13, 1625-1643.
Ganguly, A., Yang, H., Pedroza, M., Bhattacharya, R., and Cabral, F. (2011). Mitotic centromere-associated kinesin (MCAK) mediates paclitaxel resistance. J Biol Chem 286, 36378-36384.
Hearn, J.M., Romero-Canelon, I., Munro, A.F., Fu, Y., Pizarro, A.M., Garnett, M.J., McDermott, U., Carragher, N.O., and Sadler, P.J. (2015). Potent organo-osmium compound shifts metabolism in epithelial ovarian cancer cells. Proc Natl Acad Sci U S A 112, E3800-3805.