MRC DiMeN Doctoral Training Partnership: Optogenetic modelling of a hallmark neurodegeneration pathology for mechanistic research and drug discovery

Background 

The events during the initiation of pathological protein aggregation in neurodegenerative diseases of aging has to date been difficult to capture. Modulation of aggregation at this critical molecular stage offers a point of therapeutic intervention. Recent work from the primary supervisor’s laboratory shows optogenetic manipulation of protein aggregation as a promising tool reproducing cellular pathology in the neurodegenerative diseases motor neuron disease (MND) and frontotemporal dementia (FTD). Chemical-free, light-inducible aggregation of toxic dipeptide repeat (DPR) species derived from C9ORF72 gene with a repeat expansion, the most common genetic cause of MND/FTD, was for the first time achieved in cellular models. When transferred into human neuronal models and in vivo (whole-organismal) models, this platform can be used for mechanistic experiments and assay development for drug screening purposes.

Objectives

1. Establish neuronal models with inducible optoDPR aggregation to comprehensively characterise DPR aggregation-induced neuropathology.

2. Generate in vivo (Drosophila) optoDPR models to analyse behavioural, tissue and cellular phenotypes elicited by DPR aggregation.

3. Develop an optoDPR cellular assay for drug screening and orthogonal assays in in vivo models for small-molecule screening and validation studies.

Timeliness and novelty

MND/FTD represents a significant unmet medical need. >30% of inherited cases are caused by C9ORF72 repeat expansion with aggregated DPR species as a hallmark pathology.  Mechanisms by which DPR aggregates form and contribute to the disease course remain unclear. Optogenetic approaches is a state-of-the-art approach whose application in the neuropathology field holds a great modelling and screening promise but is still in its infancy.

Light-controllable protein oligomerisation systems provide a versatile tool for manipulation of protein aggregation. Primary supervisor’s team has pioneered this approach to model C9ORF72 DPR aggregation (manuscript in preparation). Transferring this technique to human neurons and a whole organism amenable to behavioural phenotyping and tissue-level studies will generate a bespoke toolkit for in-depth mechanistic interrogation of aggregation and drug screening applications.

Experimental Approach

Physiologically relevant MND/FTD optoDPR models will be generated using lentiviral transduction in human iPSC/ES cells (primary supervisor). Imaging and biochemical experiments will be performed on high-content automated imaging systems and custom light arrays, respectively. The corresponding Drosophila optoDPR models will be generated and subjected to behavioural phenotyping and imaging studies (second supervisor). Subsequent cellular assay development will be carried out under stringent industry standards, followed by screens of focused small-molecule libraries available in-house (primary/third supervisor), with subsequent hit validation in Drosophila models (second supervisor). 

Overall, this study is an interdisciplinary project that will provide bespoke training in neuronal cell culture, genetic manipulation, advanced imaging, cellular high-throughput assay development, whole organismal biology and principles of rational drug design.

Environment

Dr Shelkovnikova, the primary supervisor on this project, is a UKRI Future Leaders Fellow (senior research fellow) and a group leader at the Sheffield Institute of Translational Neuroscience (SITraN). The student will join her young vibrant lab in a world-known translational institution focusing on neurodegenerative diseases. In vivo modelling work will be performed at the lab of Prof Sweeney at the University of York who is well-known for his Drosophila modelling of neurodegenerative disease phenotypes and mechanisms. The translational aspect of the project (small molecule screening) will be supported by Prof Atack, co-director of Cardiff Medicines Discovery institute.

The project is an excellent training opportunity for a curious and ambitious candidate seeking a stimulating, flexible and diverse research environment.

For any queries and informal discussions, candidates should contact Dr Shelkovnikova in the first instance, on [Email Address Removed] 

Benefits of being in the DiMeN DTP:

This project is part of the Discovery Medicine North Doctoral Training Partnership (DiMeN DTP), a diverse community of PhD students across the North of England researching the major health problems facing the world today. Our partner institutions (Universities of Leeds, Liverpool, Newcastle, York and Sheffield) are internationally recognised as centres of research excellence and can offer you access to state-of the-art facilities to deliver high impact research.

We are very proud of our student-centred ethos and committed to supporting you throughout your PhD. As part of the DTP, we offer bespoke training in key skills sought after in early career researchers, as well as opportunities to broaden your career horizons in a range of non-academic sectors.

Being funded by the MRC means you can access additional funding for research placements, international training opportunities or internships in science policy, science communication and beyond. See how our current DiMeN students have benefited from this funding here: https://www.dimen.org.uk/blog 

Further information on the programme and how to apply can be found on our website:

https://www.dimen.org.uk/how-to-apply