BJA/RCoA non-clinical PhD Studentships

University of Leeds

Title
The use of CRISPR-Cas gene editing for functional characterisation of genetic variants associated with malignant hyperthermia susceptibility

Amount
£89,210

Scientific Abstract
Malignant hyperthermia (MH) is a potentially fatal hypermetabolic reaction affecting genetically susceptible individuals exposed to inhalational anaesthetics. MH susceptibility is associated with variants in the RYR1 gene in ~75% of cases but accumulating evidence suggests that additional genetic changes are required in at least 25% of these patients for the condition to manifest clinically. Our high-throughput genetic screening strategy has identified variants in other genes that could exacerbate mitochondrial dysfunction in MH, resulting in increased production of reactive oxygen species that amplify the effect of the RYR1 variant. This project will investigate the impact of variants in two of these other genes, PYGM and CPT2, on the phenotype of muscle cells harbouring the RYR1 variant most frequently found in the UK MH population. We will initially generate PYGM and CPT2 null myoblasts using CRISPR-Cas gene editing knock-out. Myoblasts will be expanded and differentiated to form myotubes which will then be transduced with mutant forms of PYGM or CPT2 that carry variants identified in MH patients. The impact of the mutated PYGM and CPT2 on the sensitivity of the myotubes to RYR1 agonists (caffeine and halothane) will be determined using changes of fluorescence of cells loaded with fluorescent calcium dye.

University of Aberdeen

Title
Combining omega-3 docosahexaenoic acid and melatonin to manage spinal cord injury-induced central neuropathic pain

Amount
£88,668

Scientific Abstract
Worldwide, there are 2.5 million people living with spinal injury and roughly 500,000 new cases annually. About half of spinal injury patients develop neuropathic pain, which is driven by neuroinflammation, has no effective treatments, and impacts significantly on daily functioning. Our recent studies, using novel and clinically relevant animal modelling of this pain, demonstrate for the first time that systemic treatment with omega-3 docosahexaenoic acid (DHA) attenuates neuropathic pain development after spinal injury by targeting microglial cells that play a key role in this pain. Strong evidence in rats also suggests that melatonin has anti-inflammatory properties and can attenuate microglial activation and prevent chemotherapy induced neuropathic pain. Therefore, this proposal aims to develop a novel synergistic treatment combining DHA and melatonin in order to 1) prevent neuropathic pain development after spinal injury and 2) abolish already established neuropathic pain at chronic stages of spinal injury. The project shall exploit state-of-the-art neurochemical, flow cytometry and behavioural tools in a clinically relevant animal model of spinal injury. DHA and melatonin have excellent clinical safety records. By providing a path towards an effective and safe treatment, the project will have an impact on clinical management of neuropathic pain after spinal injury.