Spectronaut16发布会讲座，缺血性肾损伤及干预质量

High-throughput DIA Analysis of Ischemic Kidney Injury and Therapeutic Interventions Using Multiple MS Platforms

Birgit Schilling (The Buck Institute)

Post-translational modifications (PTMs) dynamically regulate proteins and pathways through immediate signaling cascades and resulting changes in protein function or activity. Here, we are investigating the protective role of lysine succinylation during ischemia- and -reperfusion-induced acute kidney injury (AKI) which typically would cause kidney dysfunction and oxidative stress. Recently, we examined the succinylome profile during acute kidney injury and the effects of sirtuin-5 knockout, and alternatively the use of 2 different nutritional supplements on AKI injury recovery in mice. Using data-independent acquisitions, we identified and quantified 3,666 succinylated sites in total, and determined that the diet induced significant hypersuccinylation of 1,085 sites which remodeled the succinylome completely. However, this treatment barely affected the proteome itself. Upregulated succinylation sites were related to lipid and fatty acid oxidation, and peroxisomal metabolic pathways. We hypothesize that the increased succinylation levels promote a switch of the fatty acid oxidation from mitochondria to peroxisomes which greatly reduces oxidative stress and thereby protects against AKI. Our spectral library free directDIA workflows using the Orbitrap Eclipse are highly efficient and have allowed us to optimize and increase throughput for our PTM-DIA quantification workflows. We also investigated changes on the protein level with AKI (using a ZenoTOF 7600 system) which showed significant and broad protein level changes even with short ischemia reperfusion injury times. The novel Spectronaut 16 allowed us to improve our identification and quantification rates searching our data-independent acquisitions by 20-25 %. Altogether, our work demonstrates the promising translational and therapeutic applications of nutritional supplements to rescue mammalian (mouse, non-human primate and human) kidney injury phenotypes, and in addition to provide alternative energy sources for patients with genetic disorders affecting mitochondrial metabolisms.