Project No. 2307
Dr Yury Bogdanov – University of Southampton
Dr Rhys Morgan- University of Sussex
Gamma-aminobutyric acid (GABA) is an inhibitory neurotransmitter in the central nervous system (CNS).
It acts through ionotropic (GABA(A)receptors) and metabotropic (GABA(B) receptors).There is an emerging research field addressing the role of GABA in non-neuronal functions, notably in immune function and haematopoietic development.
It has been shown that GABA may play a role in the haematopoietic stem cell (HSC) function (Zhu et al.,2019; Shao et al.,2021; Urrutia et al.,2016). However, the origin of GABA itself and its effects on various populations of HSC is still elusive. The aim of this work is to study the expression of GABA receptors on human HSC and the impact of GABAergic signalling on normal haematopoietic development. We propose the following:
1. Identify the expression of various components of GABAergic signalling (receptors, transporters, GABA synthesis and degradation enzymes) in various developmental subsets of human cord or perhipheral blood using intracellular and cell surface staining techniques coupled to flow cytometry. Developmental subsets of blood will include expression on populations of HSC, common myeloid/lymphoid progenitors, granulocytes, erythrocyres, megakaryocytes, monocytes/macrophages, T-cells and B-cells.
2. Using an in vitro model of haematopoietic development, characterise the impact of modulating GABA receptor/component expression through lentiviral transduction/clinically approved substance treatment (alprazolam etc), on the self-renewal and differentiation potential of human HSC.
3. Evaluate the degree of crosstalk between GABA signalling and Wnt signalling in haematopoietic cells using Wnt reporter assays and anlaysis of beta-catenin expression using flow cytometry and Western blotting with nuclear/cytosol fractionation.
This work will involve various techniques of molecular and cell biology, genetic manipulation qPCR, ELISA, Western blotting and FACS.
Understanding the multifaceted role of GABA in HSC biology will open a potential of repurposing of the clinically approved substances that are used for various neurological conditions (anxiety, epilepsy, substance abuse) to treat haematological conditions.