4-year PhD Studentship: Control of neuronal differentiation and ciliogenesis through mineralocorticoid receptor (MR) – Regulatory Factor X (RFX) interaction in embryonic and adult neurogenesis

Glucocorticoid hormones (GCs) are of critical importance for health[1]. They bind to two receptor types in the hippocampus: the mineralocorticoid (MR) and glucocorticoid receptor (GR) which act as ligand-dependent transcription factors[2-4]. Recently, using ChIP-Sequencing, we identified the genes interacting with MR and GR under baseline and stress conditions[5]. Our research revealed that MR and GR interact with neuroplasticity-associated genes, supporting the concept that GCs regulate adaptive learning and memory processes. Moreover, we discovered that specifically MR interacts with genes associated with cilia structure and function, and neuronal differentiation[5]. MR interacts with these genes at RFX (Regulatory Factor X) sites which may involve co-binding with RFX factors like RFX3. Recent RFX3 ChIP-seq work indeed confirmed a high degree of overlap between binding of this factor and MR at RFX sites within ciliary genes. Functional work on human foetal neuronal progenitor cells (hfNPCs) and adult rat dentate gyrus (DG) tissue uncovered that during neuronal differentiation MR is induced in neuronal progenitor cells which is of critical importance for the differentiation and the ciliogenesis process[5; Haque S. et al., unpublished]. Presently, the underpinning mechanisms of interaction and the exact functional role of MR and RFX3 in ciliogenesis and neurogenesis is still unclear.

Aims and Objectives

We hypothesise that MR and RFX3 interact at the genomic level and, in a coordinate manner, regulate ciliogenesis and neuronal differentiation in hfNPCs and the rat DG. Our aim is to investigate the mechanistic and functional significance of MR-RFX3 interaction in these processes. Our objectives are:

• To study the genomic-level interaction of MR and RFX3 in hfNPCs and rat DG, and its implications for gene transcription, ciliogenesis and neuronal differentiation.
• To investigate the role of MR and RFX3 in chronic stress and aging where levels of adult neurogenesis are reportedly diminished.

Methodology

The student will use the following techniques: cell culturing of hfNPCs and/or iPSCs (induced Pluripotent Stem Cells), AAV/lenti-viral gene knock-down, chromatin immunoprecipitation (ChIP), RNA analysis, qPCR, RNAscope, Co-IP, Western blot analysis, immuno-fluorescence, experimental animal work, radio-immuno assays/ELISAs, bioinformatics and pathway (Gene Ontology (GO) and Ingenuity Pathway Analysis (IPA) analysis of ChIP-seq/RNAseq data. All technologies are presently running in the Neuro-Epigenetics (Reul) and Stem Cell (Cordero Llana) laboratories. The student will receive training in state-of-the-art multi/interdisciplinary skills which includes the aforementioned techniques as well as in experimental design, critical data evaluation, statistical and bioinformatics analysis, hypothesis building and mechanistic/functional network analysis.

Keywords

Neurogenesis, mineralocorticoid receptor, Regulatory Factor X, neuronal differentiation, cilium, human progenitor cells, adult eurogenesis, dentate gyrus

How to apply for this project

This project will be based in Bristol Medical School - Translational Health Sciences in the Faculty of Health Sciences at the University of Bristol.

Please visit the Faculty of Health Sciences website for details of how to apply