Possible role of mutant vimentin in cancer progression and metastasis


   Institute of Dentistry

  , ,  Applications accepted all year round  Self-Funded PhD Students Only

About the Project

Background: This project will investigate the role of the cytoskeletal protein vimentin in cancer progression and metastasis using cell and molecular biology approaches.

A form of epithelial-to-mesenchymal transition (EMT) occurs frequently in more advanced epithelial cancers and is associated with a higher degree of tumour aggressiveness. The cellular features include loss of epithelial characteristics including polarity and cell-cell adhesion, as well as a switch from an epithelial intermediate filament (IF) protein network (keratins) to vimentin (Paccione et al 2008; Kalluri and Weinberg, 2009), which is characteristic of cells of mesenchymal origin. Expression of vimentin in tumours is therefore a diagnostic marker for tumour progression and metastasis.

Vimentin is a 466 residues long type III IF protein split into three domains, the central alpha helical rod domain is flanked by non-helical head and tail domains. The head and rod domains play crucial roles in filament assembly while the tail domain determine the filament thickness and interactions with other cellular proteins (Usman et al 2022a). Earlier studies have shown that in most human cancers especially in advance stages, the expressed vimentin is mutated throughout the polypeptide. These mutations include translation termination and missense substitutions throughout the polypeptide. The rod domain is reported to contain 49 missense substitutions, with head and tail domains containing about 10 each (Usman et al 2021) . The role of these mutations in cancer progression and/or spread of tumour is not known.

Using a cell culture-based model, we have recently observed that vimentin, in the absence of endogenous vimentin, suppresses keratin expression and increases migration of cancer epithelial cells, two of the characteristics of EMT. Transcriptome analysis has shown that vimentin can downregulate endothelial cadherin, CDH5, which makes the cells freer to migrate faster (Usman et al 2022b). Vimentin expression in cancer epithelial cells can at least partially mimic EMT characteristics.

To study the effect of mutant vimentin we shall use site-directed mutagenesis to mutate those residues that are frequently mutated in more than one tumour type such as P57R, E221; A247; T266; R304; S325; R345; L380I; K390; E407 and express them in vimentin-deficient cancer epithelial cells. The effect mutant vimentin on cell proliferation, migration, invasion and self-renewal will be measured as described previously (Usman et al 2022b). The effect of mutations on gene expression will be measured by transcriptome analyses and validated by qRT-PCR, shRNA knockdown and used on mouse model.

Expected outcome: If our hypothesis that mutant vimentin can lead to cancer progression and tissue invasion is valid, it should be making it a therapeutic target in studies going forward.

Techniques to be employed: The project will use molecular cloning, 2-D and 3-D cell culture, cell migration, invasion and cell proliferation assays, immunocytochemistry, flow cytometry, fluorescence imaging including confocal microscopy, live cell imaging, western blotting, retroviral packaging and transduction, qRT-PCR, single cell and bulk transcriptome analyses , cytokine profiling, non-coding RNA profiling and animal experiments.

Admission Requirements 

A graduate with at least an upper second-class honours degree or a distinction in an experimental MSc degree is required for this PhD project. The candidate should have strong interests and preferably with some experience in cell and molecular biology, pharmacology, pharmacy, biochemistry, biotechnology or biomedical science.

If English is not your first language, the standard requirement for English is an IELTS score of 6.5 overall for non-clinical projects and 7 overall for clinical projects (or equivalent). More details about language requirements can be found here

For more information on the project, please contact Prof Ahmed Waseem (

For information on the application process, please contact  


Biological Sciences (4) Medicine (26)

Funding Notes

We will consider applications from prospective students with a source of funding to cover tuition fees and bench fees for three years full-time or 6 years part-time. Both self-funded and sponsored students will be considered.
UK nationals, Irish citizens and those with settled status under the EU Settlement Scheme or indefinite leave to remain in the UK might be eligible for a doctoral loan for both the cost of tuition fees and a yearly stipend over the course of the PhD programme from Student Finance England: View Website

References

Paccione RJ, Miyazaki H, Patel V, Waseem A, Gutkind JS, Zehner ZE, Yeudall WA (2008). Keratin down-regulation in vimentin-positive cancer cells is reversible by vimentin RNA interference, which inhibits growth and motility. Mol Cancer Ther 7:2894-903.
Kalluri R and Weinberg RA (2009) The basics of epithelial-mesenchymal transition. J Clin Invest 119: 1420–1428.
Usman, S.; Aldehlawi, H.; Nguyen, T.K.N.; Teh, M.-T.; Waseem, A (2022a). Impact of N-Terminal Tags on De Novo Vimentin Intermediate Filament Assembly. Int. J. Mol. Sci. 23, 6349.
Usman, S.; Waseem, N.H.; Nguyen, T.K.N.; Mohsin, S.; Jamal, A.; Teh, M.-T.; Waseem, A (2021) Vimentin Is at the Heart of Epithelial Mesenchymal Transition (EMT) Mediated Metastasis. Cancers 13, 4985.
Usman, S.; Jamal, A.; Bushaala, A.; Waseem, N.H.; Al-Dehlawi, H.; Yeudall, W.A.; Teh, M.-T.; Tummala, H.; Waseem, A. (2022b) Transcriptome Analysis Reveals Vimentin-Induced Disruption of Cell–Cell Associations Augments Breast Cancer Cell Migration. Cells 11, 4035.

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