C70-29 Transfer RNA Alters the Inflammatory Response to Bacterial and Viral Infection

Abstract Rationale Transfer RNA facilitates the translation of proteins by bringing amino acids to the growing polypeptide chain. Overall tRNA abundance has been demonstrated to modify translation efficiency. In addition, multiple tRNA synthetases, enzymes that attach amino acids to tRNA, have been implicated in the inflammatory response. tRNA modifying enzymes have also been shown to alter innate immune signaling. However, to our knowledge, there have been no direct studies examining how the abundance of transfer RNA regulates the inflammatory response. Methods To determine how tRNA modifies the response to infection, we created a tRNA expression library encompassing 150 tRNA isodecoders under their endogenous promoters. In addition, we created vectors expressing 22 mitochondrial tRNA with adaptor sequences previously demonstrated to ensure proper mitochondrial localization and processing. Using nanopore sequencing, we confirmed that individual tRNA isodecoders were increased following transfection. In a high-throughput assay, we examined the effect of individual tRNA on cell viability and cytokine production in response to various inflammatory stimuli including influenza and bacterial infection. Results We determined that overexpression of multiple tRNA significantly altered cell viability and the production of inflammatory cytokines in response to multiple stimuli. Thes findings were recapitulated in primary cell models. Conclusions Our results suggest that altering the expression of both cellular and mitochondrial tRNA modifies the inflammatory response and cell viability in response to infection. We will utilize this platform to examine the role of tRNA in response to multiple inflammatory stimuli including bacterial and viral infection. By understanding how individual species of tRNA modify cellular processes, we hope to identify new biomarkers or therapeutic avenues. This abstract is funded by: 5R01HL169586