Deciphering the metabolic requirements of tissue resident T cells

The aim of the project is to understand the gene-metabolism interactions affecting tissue-resident T cell migration, phenotype, differentiation, and function. This will be assessed through a loss-of-function CrispR screen in T cells isolated from mouse splenocytes, targeting metabolically relevant genes with the aim of highlighting potential metabolic gene-essentiality. This will allow for the assessment of the relative impact of metabolic deficiencies on different T cell subtypes (CD4, conventional T cells (T conv), CD8, regulatory T cells (T reg), and naïve/activated/memory subsets) within the circulating lymphoid compartment to identify cell type-specific requirements.

The project will require the utilisiation of the labs expertise and unique cell tracking system: Flowcode system. This established system can be used to track immune cell migration relative to WT T cells using flow cytometry to not only compare potential differences in tissue residency but also phenotype the cells to gain a holistic understanding of the gene function.

Following initial unbiased screening, hits are to be validated and screening repeated in pathological contexts such as brain injury, COPD or gut inflammatory conditions to assess gene importance in various contexts. To understand the mechanisms by which any hits elicit the observed effects from the screen further analysis will be done, using techniques such as RNA sequencing, Gene ontological analysis, and in vitro assessment of cellular function following gene knock-down. This can also be paired with techniques like LC-MS to understand the tissue microenvironment and see if there is any contribution from the cells immediate environment to the effects of the gene knockdown.


This will enable us to gain a more in-depth understanding of the immunometabolism of T cells, and highlight potential therapeutic avenues to explore in metabolically challenged tissues and pathology, where metabolic rewiring and perturbations are often seen.