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Defining the role of WT1 in cellular and molecular processes during gastrulation, in embryos and gastruloids


   Faculty of Health and Life Science

   Tuesday, February 21, 2023  Self-Funded PhD Students Only

About the Project

In an ageing population, injury and degenerative processes lead to increased pressure on the NHS due to demand on treatments. One of the key health problems in a Western ageing society is cardiovascular disease, involving vascular smooth muscle cells which can degenerate in various disease and injury situations, resulting in atherosclerosis, arterial stiffening, and hypertension. To help alleviate this pressure, novel, more efficient and better targeted treatments are required. Because tissue development and regeneration share common mechanisms, a better understanding of how developmental processes are regulated, can provide important mechanistic cues for regenerative therapies to treat degenerative disease of the vascular wall.

In recent years, our research has studied the role of the transcriptional regulator Wilms' tumour 1 protein (WT1) in the development and maintenance of tissues and organs. WT1 is a critical modulator of key developmental processes in vertebrates including the formation of the mesoderm-derived organs: kidney, spleen and pancreas. We have used genetic lineage tracing experiments in genetically modified mice to investigate the role of WT1 in tissue development. Our data suggest a novel developmental role for WT1 during gastrulation stages, regulating the formation of mesoderm-derived vascular smooth muscle. However, due to the difficulty of studying Wt1 during gastrulation in vivo, we still do not have a nuanced understanding of the molecular and cellular mechanisms of how Wt1 modulates cells towards a vascular smooth muscle fate. To study these processes, we will take a hybrid approach, combining in vivo studies of mouse embryos with a state-of-the-art in vitro approach using gastruloids. Gastruloids are small aggregates of mouse embryonic stem cells grown in 3D that mimic many of the morphological and patterning processes that take place during gastrulation, allowing us to study how Wt1 modulates developmental patterning processes in vitro.

In this project, we aim to generate ESCs from genetically modified, compound mutant mice, which will allow inducible lineage tracing of WT1-expressing cells, or knock-out of WT1. Based on these, we will establish the lineage of WT1-expressing cells by comparing gastruloids with mouse embryos. This will involve developing live cell imaging and ex vivo culture protocols using cutting edge technologies including light sheet and confocal microscopy. Lastly, we will determine the importance of WT1 for cellular patterning events during gastrulation, using the established technologies of live cell imaging and ex vivo culture. In addition, we will apply flow cytometry and RNAseq to determine the molecular role of WT1 in these patterning processes.

This studentship will utilise state-of-the-art technology and world-leading equipment and facilities, bringing together a unique opportunity for a student to analyse fundamental mechanisms of early embryonic development that are relevant for tissue regeneration processes in vascular smooth muscle cells of the cardiovascular system.

The successful student will gain training in a multitude of techniques, including immunofluorescence staining, qPCR, flow cytometry and protein expression analysis (Western etc), mouse genetics, analysis of mouse embryos, ES cell culture, culture of gastruloids, microscopy, image analysis and introduction to biostatistical approaches and data handling including R.

The research project will be based in the Department of Molecular Physiology and Cell Signalling at the University of Liverpool.

For any enquiries please contact Dr Bettina Wilm, 

Application is by CV and covering letter.  The covering letter must detail your interest in the studentship, related experience and training and suitability for the position.  Applications should be sent to Dr Bettina Wilm,

Funding Notes

We are looking for a self-funded student or a student who have secured funding from an independent body. There is no financial support available from Liverpool for this study. Please see website for PhD student fees at the University of Liverpool: View Website

The successful applicant will be expected to have funding in place for the tuition fees (check University of Liverpool website), consumables/bench fee (£ 18,000 per annum) and living expenses during their stay in Liverpool.


Funding Notes

This is a self funded opportunity.
A £2000 ISMIB Travel and Training Support Grant may be available to new self-funded applicants.

References

1. Wilm et al. (2005) The serosal mesothelium is a major source of smooth muscle cells of the gut vasculature. Development 132(23):5317-5328. Doi: 10.1242/dev.02141
2. Held et al. (2018) Ex vivo live cell tracking in kidney organoids using light sheet fluorescence microscopy. PLoS One 26;13(7):e0199918. doi: 10.1371/journal.pone.0199918.
3. Beccari et al. (2018) Multi-axial self-organization properties of mouse embryonic stem cells into gastruloids. Nature 562, pp 272–276 DOI: 10.1038/s41586-018-0578-0

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