Investigating epilepsy associated tRNA fragments

This project aims to investigate the generation and function of tRNA fragments in neuronal cell models of epilepsy. It is vital to understand the molecular pathways that lead to generation of tRNA fragments as these may reveal novel therapeutic targets for the prevention of seizures. 

In 2019 we published the first example of a blood-based biomarker for seizure imminence (Hogg et al, 2019, JCI). We described a signature of 3 tRNA fragments which were elevated in plasma collected in advance of seizures and returned to baseline levels in post-seizure samples. We demonstrated that these tRNA fragments are generated in neurons and that levels are regulated in response to neuronal activity. More recently, tRNA fragments in blood have been reported as biomarkers in a number of neurological syndromes and cancers. tRNA fragments are a novel class of non-coding RNA with wide ranging reported effects, however little is known about their function in neurons.  

The project will use neural cell lines and induced pluripotent stem cells differentiated into cortical neurons to model epilepsy and epileptogenesis in vitro. We will use pharmacological agents to stimulate neuronal activity and investigate how these influence tRNA fragment levels. We investigate the ribonuclease(s) responsible for generating tRNA fragments and analyse the effect of depletion on neuronal activity. To model epileptogenesis we will induce a range of physiological stresses and analyse the effect on tRNA fragment levels. We will assess the role of tRNA fragments in neuronal survival following physiological stress. We will determine factors interacting with tRNA fragments and investigate the secretion pathway and encapsulation status of tRNA fragments. Together this project will provide a comprehensive characterisation of neuronal tRNA fragments and how they are influenced by neuronal activity. 

The successful applicant will gain experience in a wide range of techniques including cellular and molecular biology, differentiation and maintenance of neuronal cultures, standard biochemistry techniques, RNA and protein analysis techniques, immunocytochemistry, live cell imaging and image analysis techniques. The student will join a vibrant neuroscience research environment at NTU with expertise in a range of different models and scientific approaches. 

This project offers a great opportunity to join the newly established neuroscience research theme within the Centre for Healthy Aging and Understanding Disease based at NTUs Clifton campus.