The skin and other epithelial surfaces, such as the oral mucosa, play a critical role as a barrier to pathogens and other environmental insults. This renders these surfaces vulnerable to injury, necessitating efficient wound healing responses. These responses can be affected by many factors, including diseases such as diabetes, as well as cancer therapy and age. Defective wound healing is a major cause of ill health, poor quality of life and poses a huge economic burden on the NHS. Peripheral neuropathy, the loss of neurons from the connective tissue underlying epithelial barriers, is strongly implicated in defective wound healing. The mechanisms underlying this, however, remain poorly understood.
We have evidence that fibroblasts, the predominant cell type found in connective tissue, are stimulated to differentiate into myofibroblasts, a process critical for effective wound healing, by signals generated by sensory neurons. We have also observed that isolated primary neurons are activated and stimulated to form neurites by myofibroblast-derived signals. In this project, we propose to comprehensively characterise the interactions between sensory neurons and fibroblasts, with the aim of elucidating fundamental mechanisms underlying the influence of neurons on wound healing and diseases such as cancer.
Specifically, we will test the hypothesis that fibroblast:neuronal interactions are necessary for effective wound healing. The following specific aims, in the timescales proposed, will be addressed:
Aim 1. Generate trigeminal ganglion (TG) neuronal cultures and characterise the response of fibroblasts to TG-derived factors. This will be achieved using techniques including RNAseq, qPCR, western blot and immunocytochemistry for known myofibroblast markers (eg α smooth muscle actin) and bioinformatic analyses.
Aim 2. Determine which cells in the TG are involved in cross-talk with fibroblasts. The cellular composition of TG cultures will be assessed by immunocytochemistry and FACS cell sorting using markers of relevant cell types (eg neuronal, glial, smooth muscle and endothelial cells), and responses to fibroblast-derived signals assessed by calcium flux imaging, capitalising on the expertise of the co-supervisor Anton Nikolaev, RNAseq (as described in aim 1), and neurite outgrowth assessed by microscopy.
Aim 3. Assess the influence of neuronal:fibroblast interactions on wound healing in vivo. We have preliminary evidence of defective wound healing in vivo in an experimental model of denervation. Here, we will extend these findings to assess the abundance and phenotype of fibroblasts in innervated and denervated wounds. In addition, we will examine the effect of neuronal factors on wound healing, using fibrin glue containing neuronal mediators and specific inhibitors previously optimised in vivo in Prof Boissonade’s laboratory.
Expected outcomes: This project will generate insight into the roles of fibroblast:neuronal interactions in the context of wound healing, and identify molecular pathways which may be amenable to therapeutic intervention. In addition, the potential of neuronal mediators as pro-regenerative agents will be determined, providing a platform for subsequent translational studies.
Environment: This project is a collaboration between Prof Lambert and Boissonade’s groups in the School of Clinical Dentistry, and Dr Nikolaev’s group in Biomedical Sciences. All three groups offer a vibrant environment for PhD students, with full training provided and access to cutting edge equipment and expertise. In addition, as a Neuroscience Institute-funded student, you will have access to the Institute’s facilities, a world-leading group of Neuroscience researchers, and training and career development opportunities provided by the Institute.
Applications are open to students from both the UK and overseas, though we note that due to funding constraints the availability of positions for students with overseas fee status will be more limited. We anticipate competition for these studentships to be very intense. We would expect applicants to have an excellent undergraduate degree in a relevant discipline. We would also expect applicants to have completed or be undertaking a relevant master’s degree to a similar very high standard (or have equivalent research experience).
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Please complete a University Postgraduate Research Application form available here: https://www.sheffield.ac.uk/postgradapplication/
Please clearly state the prospective main supervisor in the respective box and select ‘Neuroscience’ as the department.
After the application closing date, we will shortlist applicants for an online interview. We expect to carry out interviews (each lasting approximately 30 minutes) on Tuesday 27th April (am, GMT) and Tuesday 4th May (pm, GMT). If you are shortlisted for interview, we will aim to inform you of this no later than the end of Friday 23rd April. If you are unable to attend at the specified times, please let us know if we confirm that we would like to interview you.
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