CHINA SCHOLARSHIP COUNCIL: Vaulting the void: The vault particle as a novel mediator of intercellular communication in the tumour microenvironment

Details

Background

The vault particle is the largest known eukaryotic ribonucleoprotein complex. It is composed of 78 copies of major vault protein (MVP) as well as two associated proteins (TEP1 and PARP4) and vault RNA (vtRNA) [1,2].

The function of the vault particle remains elusive. However, it has been implicated in intracellular transport and multidrug resistance. VtRNA have been linked to resistance to apoptosis and chemoresistance, independent of the vault particle [3].

Cancer cells release a complex mixture of soluble factors and extracellular particles into the tumour microenvironment (TME), which have been implicated in driving tumorigenesis. In recent decades, extracellular vesicles (EVs) have attracted attention due to their role in intercellular communication in the TME. MVP and vtRNA have repeatedly been reported as EV cargo. However, a recent report [4] (and our data https://www.biorxiv.org/content/10.1101/2023.11.09.566362v1) demonstrated the presence of intact vault particles in the extracellular space. We have shown that vault particles are frequently co-isolated with EVs.

Experimental approach

We will utilise the state-of-the-art NanoAnalyzer to characterise the extracellular particles released from wild-type oral cancer cells and a vault particle deficient cell line (already generated in house). You will receive full training to use the NanoAnalyzer by dedicated technical staff. You will also become proficient in cell culture, enrichment of extracellular particles (using techniques such as size exclusion chromatography and ultracentrifugation) and fluorescent labelling of extracellular particles.

We will investigate the mechanism of vault particle release using advanced microscopy techniques such as confocal microscopy and transmission electron microscopy coupled with immunogold labelling. We will use small interfering RNA and CRISPR Cas9 gene editing to perturb putative export machinery. You will also utilise quantitative PCR and western blotting to analyse gene expression and protein abundance, respectively.

We will use in vitro cell culture to investigate if transfer of vtRNA via uptake of vault particles can endow an apoptosis resistant phenotype in cells that would be found in the TME. Vault particles will be tagged by genetic manipulation to allow tracking of release and uptake.

Supervisor one: Dr Stuart Hunt

https://www.sheffield.ac.uk/dentalschool/our-people/academic-staff/stuart-hunt

https://twitter.com/Dr_Stuart_Hunt

Supervisor two: Prof Dan Lambert

https://www.sheffield.ac.uk/dentalschool/our-people/academic-staff/daniel-lambert

Entry Requirements:

Candidates must have a first or upper second class honours degree or significant research experience. 

To be eligible for a CSC scholarship, you must:

• be applying to start a new full-time PhD programme at Sheffield
• have a strong academic background
• be a national of and reside in, mainland China (not including Hong Kong or Macau) – see CSC policies
• intend to return to China once your programme is completed
• hold a study offer from the University of Sheffield (see below for further details)
• satisfy all of the CSC's selection criteria 

How to apply:

Please complete a University Postgraduate Research Application form available here: https://www.sheffield.ac.uk/postgraduate/phd/apply/applying

Please clearly state the prospective main supervisor in the respective box and select ‘School of Clinical Dentistry’ as the department.

Proposed start date - October 2024

For further information on the CSC scheme please see - https://www.sheffield.ac.uk/postgraduate/phd/scholarships/csc