Novel cellular approaches to investigate the relation between energy metabolism and vascular function

Supervisor 1: Dr Sébastien Serres

Supervisor 2:Dr Andrew Bennett and Dr Tim Constantin-Teodosiu

Application deadline:Open all year round

Project Title:novelcellular approaches to investigate the relation between energy metabolism and vascular function

Project Description:

Background:Blood supply of oxygen and glucose to the cells is regulated by essential mechanisms that ensure normal functioning of vital organs. In the brain, a neurovascular coupling mechanism increases local cerebral blood flow (CBF) to match metabolic demand upon cerebral activity1. Although efforts have been done in defining how brain cells such as astrocytes and pericytes control CBF2, we still do not know how metabolic changes in these cells alter the control of CBF in disease.

Hypothesis:From our recent work, we have shown that extracellular matrix (ECM) receptors that attach astrocytes to blood vessels are crucial for CBF response and this is reduced when astrocyte are activated in response to disease. As activated astrocytes undergo metabolic changes, we hypothesise that this may modulate the expression of ECM receptors, which in turn, disrupts the control of CBF.

Aim:In this project, you will use a combination of cell culture, molecular biology and immunostaining approaches, together with novel imaging methods to identify and assess metabolic pathways in astrocytes that are associated with the regulation of ECM receptors.

Project:Firstly, you will focus on developing an in vitrocell culture assay using animal and human tissues to characterise the expression of ECM receptors (e.g. dystroglycans) on astrocytes in both normal and activated condition. In a similar setting, you will investigate which metabolic pathways are dysregulated in activated condition, and thuscan be targeted for shRNA inhibition. This approach will enable to identify metabolic pathways that control the expression of ECM receptors in cultured astrocytes.

Secondly, you will focus on developing an in vitro 3D culture assay consisting of astrocytes, smooth muscle and endothelial cells to assess metabolic pathways using Dynamic Nuclear Polarisation (DNP) 13C MRS. This method has revolutionised metabolic studies in animal models by enabling the rapid and sensitive detection of 13C-labelled cellular metabolites in vivo and, more importantly, their enzymatic transformation into other species. In collaboration with the Sir Peter Mansfield Magnetic Resonance Centre (SPMIC) based at the University of Nottingham, youwill be the first to use DNP13C MRSto investigatebrain metabolismin 3D cultureassay. This work will be used as a platform to evaluate hyperpolarized 13C-labeled precursors for metabolic studies in both animal models and human subjects, in which the successful applicant will be involved.

Summary: the work carried out in this PhD will provide expertise in cell culture,immunostaining, metabolic assays andadvanced DNP 13C MRS methodstoinvestigate the role of cellular metabolism in ECM-receptorexpression on astrocytes. Both experiments and data analyses will be done in collaboration with research groupsbased at the University of Nottingham.This project aims to identify metabolic pathways that control ECM- receptor expression and also optimise a cutting-edge method for metabolic imaging.

1.Roy CS, Sherrington CS. On the Regulation of the Blood-supply of the Brain. J Physiol11, 85-158 117 (1890).

2.Attwell D, Buchan AM, Charpak S, Lauritzen M, Macvicar BA, Newman EA. Glial and neuronal control of brain blood flow. Nature468, 232-243 (2010).

Funding Notes: This project will be funded by the School of Life Sciences at the University of Nottingham, UK.

Students must have a UK 2.1 degree (or equivalent) in a relevant subject (e.g. neuroscience, biochemistry). The studentship will include the payment of tuition fees at the Home/EU rate and a maintenance stipend per annum

Project classification:

  • Brain cell culture
  • Brain metabolism
  • Vascular function
  • Dynamic Nuclear Polarisation
  • 13C Magnetic Resonance Spectroscopy

Application Enquiries to:

Dr Sébastien Serres