About the Project
This proposal aligns with the industrial strategy challenge in the sectors of leading-edge healthcare and medicine, and bioscience and biotechnology, through aiding and enhancing the development of a new class of pharmaceuticals, RNA-based biologics, in conjunction with our partner organisation AstraZeneca (AZ). Recent advances in RNA biology have led to an expansion of new therapeutic avenues to treat a range of diseases. In particular, in vitro transcribed modified mRNAs (IVTmRNAs) are currently under development by AZ who have a major investment in this area. However, there are number of issues associated with these new biologics, which currently restricts their use and further development. One major concern is that inappropriate expression of IVTmRNAs leads to cell/tissue stress, inflammation and “off target” toxicity. The proposed project, using samples and tissues provided by AZ, aims to resolve these issues by RNA engineering to explore and design out features that induce cell stress. In addition, the proposal supports the industrial strategy by helping to close the productivity gap. Thus through training in the Unit the PhD student will gain transferable skill sets and equally importantly, bespoke training in toxicology, which is an area where the UK lacks expertise. At present there are insufficient numbers of PhD-level trained researchers in Toxicology to fill the positions available, and our industrial partners find it difficult to recruit the right calibre of individuals in this area. This proposal will therefore address a key skills gap within the UK and remove barriers to commercialisation of world-leading science carried out in this area and drive growth of the economy in this sector.
The use of in vitro transcribed modified mRNAs (mIVTmRNAs) holds promise for the safe, transient delivery of transgenes. However, there are limitations in the efficacy of IVTmRNAs due to inflammatory reactions and relatively low levels of transgene expression. This is due to antiviral mechanisms that are mediated via the delivery of naked mRNAs. To devise new strategies to overcome these issues and examine the complex dose:response relationship between mRNA burden and protein expression in single cells, we will develop i) multiplex immunohistochemical assays to detect mIVTmRNAs by in situ hybridisation (ISH), quantification of the transgene product by immunohistochemistry (IHC), and further IHC to simultaneously quantify stress response proteins. ii) Use ISH and IHC to examine the induction of inflammatory signalling pathways. iii) Examine the tissue cellular immune response by assaying for markers for transfected mRNA/protein with immune cell types. These assays will be applied to experimental tissues supplied by AstraZeneca. Based on these data we will iv) optimise mIVTmRNAs including by modifying nucleotide content, altering codon optimisation, and modification of mRNA stability elements/initiation efficiency modifiers. Treated model tissues will be supplied and re-examined and information gained will enhance the transfer of IVTmRNA technologies into a clinical setting.
Funding Notes
BBSRC eligibility rules apply.
This is a fully funded studentship to include fees, stipend, consumables and a travel / conference allowance.
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