ABSTRACT
Immune cells respond to environmental cues by way of receptors that trigger intracellular signalling cascades. These signalling cascades often involve transient protein-protein interactions. Detailed understanding of how and where in the cell intracellular proteins interact is required to understand how immune cells can be controlled and manipulated therapeutically. The function of numerous signalling proteins have been elucidated using traditional wet-lab methods. But there are many receptors for which the exact function and role remain uncertain. The expansion of large, high quality scRNA-seq databases following the Covid-19 pandemic have increased the applicability of data mining to answer outstanding questions in immunology. T-cell specific adaptor (TSAd), encoded by the SH2D2A gene, is an adaptor enriched in T and NK cells. Several interactors of TSAd, including the tyrosine kinases Lck and Itk, have been identified. However, what roles these interactions play in T and NK cell function remains unclear. To pick apart the TSAd interactome we have delved into available scRNA-seq datasets to determine genes and cell surface molecules that are concomitantly expressed with TSAd, making use of paired healthy and diseased samples to give a multi-faceted picture of the TSAd interactome. New interactors and the signals that control these interactions will be verified in genetically modified T or NK cells using classic wet-lab techniques such as pulldowns, as well as newer techniques such as the proximity ligation assay (PLA), with pilot work carried out on the established TSAd and Lck pair as a proof-of- concept.