Epigenetic control of transcription factor regulation of adult neurogenesis in the dentate gyrus

In the adult brain, new neurons are continuously generated in the hippocampal dentate gyrus (DG). This neurogenesis process is highly regulated and consists of cell proliferation, neuronal differentiation, neuronal maturation and survival. Ultimately, the new granule neurons integrate into the hippocampal neurocircuitry and become functionally implicated in cognition and mood regulation. The underlying mechanisms controlling adult neurogenesis are, however, still unclear.

We recently discovered that critical steps in the adult neurogenesis process are governed by the transcription factors MR (mineralocorticoid receptor), CREB (cAMP response element binding protein) and RFX3 (regulatory factor X 3)[1,2]. Our research showed that these transcription factors interact with key genes determining neuronal lineage commitment and progression of neuronal differentiation, including ciliogenesis, to establish new-born young DG neurons. Moreover, we recently revealed that distinct epigenetic modifications control the access of MR and GR (glucocorticoid receptor) to the hippocampal genome[3,4]. As epigenetics is known to be implicated in neurogenesis, we have postulated that access of MR, CREB and RFX to neurogenesis-relevant genes is governed by epigenetic modifications, like DNA methylation[3] and histone modification[4,5] changes. This PhD studentship will investigate the interplay between these transcription factors and the role of epigenetic mechanisms in adult DG neurogenesis in vivo and in primary progenitor cell cultures and human iPSCs in vitro. This research will advance our understanding of the molecular and cellular basis of adult neurogenesis which will benefit development of future treatments of neurological and mental disorders including neuro-engineering of nervous tissue.

Aims and objectives

General aim

To gain insight into the epigenetic control of transcription factor regulation of adult DG neurogenesis.

Hypotheses

• Adult DG neurogenesis is governed by the coordinated genomic interaction of MR, pCREB and RFX3 in the hippocampal dentate gyrus
• Accessibility of these transcription factors is determined by distinct epigenetic modifications
• Sex differences in adult DG neurogenesis are the result of distinct epigenetic and transcription factor regulation of this process

Objectives

• To investigate the genomic interaction of pCREB, MR and RFX3 at distinct phases of the adult DG neurogenesis process in vivo.
• To study the role of DNA methylation and histone modifications such as the phosphorylation and acetylation of histone H3 in the access of these transcription factors to the genome, its gene transcriptional implications and consequences for progression of the neuronal differentiation process
• To study sex differences in these epigenetic and transcription factor regulatory mechanism of adult DG neurogenesis
• To investigate these molecular and cellular mechanisms in primary progenitor cultures and iPSCs in vitro to obtain deeper insights into these complex processes.

Methodology

The student will use the following techniques: cell culturing and differentiation of human iPSCs (induced Pluripotent Stem Cells), AAV/lenti-viral gene knock-down, chromatin immunoprecipitation (ChIP; [1,6]), 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/RNA-seq 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.

Apply for this project

This project will be based in Bristol Medical School - Translational Health Sciences.

Please contact brms-pgradmin@bristol.ac.uk for further details on how to apply.

Apply now!