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International 3Rs Prize now open for applications. £30k prize (£2k personal award) for outstanding science with demonstrable 3Rs impacts.

NC3Rs | 20 Years: Pioneering Better Science
PhD Studentship

Find the target of valproic acid: pioneering the use of a non-animal model for basic biomedical (epilepsy) research

Professor Robin Williams holding up a beaker in woodlands

At a glance

Completed
Award date
September 2014 - March 2018
Grant amount
£90,000
Principal investigator
Professor Robin Williams

Co-investigator(s)

Institute
Royal Holloway University of London

R

  • Replacement
Read the abstract
View the grant profile on GtR

Overview

Aims

This project aims to build on previous NC3Rs-funded research to demonstrate the utility of an amoeba-based model to identify how valproic acid regulates PIP3 as a possible mechanism for its antiepileptic action.

Background

It has recently been shown with NC3Rs funding that a commonly used antiepileptic drug, valproic acid, blocks the turnover of phosphoinositides in a simple model organism, the amoeba Dictyostelium. It was subsequently demonstrated in the rat that seizures reduce hippocampal phosphatidylinositol trisphosphate (PIP3) and that this can be restored by valproic acid suggesting that the antiepileptic action of this drug may be exerted via PIP3 signalling. Many mechanisms for the activity of valproic acid have been proposed but an involvement in phosphoinositide signalling could explain a number of these. However, the detailed target of this effect is not known and while mechanistic studies in rats would require large numbers of animals the availability of the Dictyostelium model provides a viable alternative.

Research details and methods

This project will develop sensitive methods to analyse the effects of valproic acid on phosphoinositide turnover, and identify the specific enzyme targeted by valproic acid through use of gene knockouts and mutations. The project will expand the use of Dictyostelium as a model for study of phosphoinositides and epilepsy by heterologous expression of human proteins from this pathway. 

Application abstract

Epilepsy research generally employs a large number of rodents to use either as a source of neurons or to re-create seizures in moderate to severe experiments. The goals of these experiments are to identify how seizure treatments block seizures and to develop better treatments. We have recently shown that the most commonly used seizure control treatment, valproic acid, blocks the turnover of a specific family of chemicals (called phosphoinositides) in the simple nonanimal biomedical model, Dictyostelium. We subsequently found that valproic acid plays a similar role in the brain of rodents during seizures. Investigating how valproic acid acts in this role will require the use of hundreds of animal lives, and will involve a range of moderate to severe experiments. In this project, we will replace these animal experiments using Dictyostelium. In pursuit of this, we will develop a sensitive method to analyse the role of valproic acid in controlling phosphoinositides turnover, we will identify the specific enzyme targeted by valproic acid for this effect, and we will characterise the effect of valproic acid treatment on phosphoinositide regulation. We will also use this simple amoeba as a pioneering new system to produce human enzymes for research purposes, providing a long-term model for biomedical research into human protein function as an alternative approach to the rapidly expanding number of transgenic animals used in research.

Impacts

Publications

  1. Kelly E et al. (2018). Diacylglycerol kinase (DGKA) regulates the effect of the epilepsy and bipolar disorder treatment valproic acid in Dictyostelium discoideum. Disease Models & Mechanisms 11(9) doi: 10.1242/dmm.035600
  2. Otto GP et al. (2016). Employing Dictyostelium as an Advantageous 3Rs Model for Pharmacogenetic Research. In: Jin T., Hereld D. (eds) Chemotaxis. Methods in Molecular Biology, vol 1407. Humana Press, New York, NY doi: 10.1007/978-1-4939-3480-5_9
  3. Otto GP et al. (2016). Non-Catalytic Roles of Presenilin Throughout Evolution. Journal of Alzheimer's disease 128:54-62. doi: 10.3233/JAD-150940
  4. Williams RSB and Bate C (2018). Valproic acid and its congener propylisopropylacetic acid reduced the amount of soluble amyloid-β oligomers released from 7PA2 cells. Neuropharmacology 128:54-62. doi: 10.1016/j.neuropharm.2017.09.031