The tumour microenvironment (TME) encompasses the diverse cellular and extracellular components surrounding and interacting with tumour cells, including immune cells, fibroblasts, blood vessels, signalling molecules, and the extracellular matrix. During cancer progression, cancer cells are subject to numerous external stimuli from the TME, including mechanical cues such as increased matrix stiffness. Through mechanosensitive pathways, cancer cells sense changes in their mechanical environment and adapt their behaviour including proliferation, migration, and invasion (Zhou et al., 2022). Understanding the mechanosensing mechanisms in cancer can reveal new therapeutic targets for inhibiting metastasis and tumour growth.
The endoplasmic reticulum (ER) is a critical organelle which through a network of cisternae and tubules connects the nucleus (it is continuous with the nuclear envelope) to the plasma membrane. The ER is involved in numerous cellular functions, including protein synthesis, lipid metabolism, and calcium homeostasis and recent research, including work performed in our group, has highlighted the ER's role in mechanosensing (Guadagno et al., 2020; Song et al., 2024). In more detail, work from our group and others showed that ER tubules extend towards the plasma membrane to establish contacts with focal adhesion and regulate cell motility (Guadagno et al., 2020, Siegfried et al., 2024). The aim of this project is to further characterise the proteins interacting with the ER at focal adhesions to better understand their role, and that of the ER more generally, in mechanosensing.
In this project, the student will use state of the art imaging techniques to further dissect the role of the ER in responding to mechanical cues using cancer cell lines. The student will make use of the confocal fluorescence microscopes in the Norwegian Molecular Imaging Consortium (NorMIC) core facility at IBV to image the ER and other fluorescently tagged proteins, and biochemical assays to further validate results.
Methods:
This project will use techniques in biochemistry, cellular and molecular biology, including:
- recombinant DNA technology
- protein expression
- Western blotting
- cell culture
- cell transfection
- RNA interference
- state-of-the-art microscopy techniques for immunofluorescence and live cell imaging
- quantitative image analysis using software such as ImageJ, Imaris, Napari etc
Work place and environment:
The project will be performed at the Department of Biosciences in the group of Cinzia Progida under the daily supervision of postdoc Jonathan Townson.
The group consists of researchers at different stages of their careers (Master, PhD students, post-docs and an experienced technician).
If any questions, contact: c.a.m.progida@ibv.uio.no or j.m.townson@ibv.uio.no
Phone: 22854441
Room: 3626
References:
- Guadagno, N.A., Margiotta, A., Bjrnestad, S.A., Haugen, L.H., Kjos, I., Xu, X.C., Hu, X., Bakke, O., Margadant, F., and Progida, C. (2020). Rab18 regulates focal adhesion dynamics by interacting with kinectin-1 at the endoplasmic reticulum. J Cell Biol 219.
- Siegfried, H., Farkouh, G., Borgne, R. Le, Pioche-Durieu, C., Laplace, T. D. A., Verraes, A., Daunas, L., Verbavatz, J.-M., & Heuzé, M. L. (2024). The ER tether VAPA is required for proper cell motility and anchors ER-PM contact sites to focal adhesions. ELife, 13, e85962.
- Song, Y., Zhao, Z., Xu, L., Huang, P., Gao, J., Li, J., Wang, X., Zhou, Y., Wang, J., Zhao, W., Wang, L., Zheng, C., Gao, B., Jiang, L., Liu, K., Guo, Y., Yao, X., & Duan, L. (2024). Using an ER-specific optogenetic mechanostimulator to understand the mechanosensitivity of the endoplasmic reticulum. Developmental Cell, 59, 1–14.
- Zhou, H., Wang, M., Zhang, Y., Su, Q., Xie, Z., Chen, X., Yan, R., Li, P., Li, T., Qin, X., Yang, H., Wu, C., You, F., Li, S., & Liu, Y. (2022). Functions and clinical significance of mechanical tumor microenvironment: cancer cell sensing, mechanobiology and metastasis. Cancer Communications, 42(5), 374–400.