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Ischemia/reperfusion kidney injury: 1D and 2D LC-MS differential protein expression analysis of urine from patients undergoing CBP surgery

ABSTRACT: Urine was collected from 10 patients undergoing cardiac bypass (CBP) surgery following anesthesia induction. The study population divided into 2 post-operative outcomes: 5 experiencing acute kidney injury (AKI), and 5 not. A 1D LC-MS of individual patients was performed, as well as 2D LC-MS of pools of the five patients in each outcome set. Differential protein expression between outcomes indicated that within our experimental design, urine proteomics could not indicate AKI outcomes prior to CBP induction. SOMAscan assays on patient serum collected at the same time-point provided additional support to the notion that AKI outcomes are potentially a response to surgical stress, not a predetermined mechanism accessible prior to surgery.

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: John Wilkins

PROVIDER: MSV000081129 | MassIVE | Wed May 31 05:52:00 BST 2017

REPOSITORIES: MassIVE

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Project description:Objectives: Secretion of extracellular vesicles (EV) and associated micro-RNAs (miR) is altered during cellular stress and may serve as biomarkers of organ injury. We hypothesized that measuring changes in urinary levels of EV and miR will predict the onset of acute kidney injury in cardiac surgery patients. Design: Predictive accuracy biomarker study performed in the cohort of the REVAKI-2 trial Setting: Single center ICU between September 2015 and September 2018 Interventions: Intravenous sildenafil citrate 12.5 mg kg-1 over 150 min or dextrose 5% at the commencement of surgery. Measurements and main results: Urine samples were collected before and 24 hours after the procedure from 93 cardiac surgery patients. Urine EV concentrations and size distribution were assessed using NanoSight. EV derivation and levels were measured using flow cytometry. Samples from 10 selected patients were sequenced to detect differentially expressed miR. Verification was performed with advanced TaqMan assays in samples from all patients. Urine EV concentrations significantly increased in patients with AKI after surgery, with the percentage of EV positive for aquaporin-2 and β1-integrin also increasing. Pre surgery podocalyxin-positive EV were significantly lower, and β1-integrin EV werehigher in patients with AKI. The levels of the former correlated with the severity of theinjury. miR-125a-5p was expressed at higher levels in urine from patients with AKI stage 2/3. Levels of miR-10a-5p decreased after surgery in AKI patients; its levels correlated with the severity of the injury. Preoperative levels of podocalyxin EVs and miR-125a-5p had moderate AKI predictive value and, in a logistic model together with ICU lactate levels, offered good (AUC = 80.9%) AKI prediction. Conclusions: Lower levels of podocalyxin-positive EV at baseline predict the severity of post-surgery AKI. Urine EV concentrations and miR expression offer excellent predictive accuracy when combined with commonly measured biomarkers.

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Ischemia/reperfusion kidney injury: 1D and 2D LC-MS differential protein expression analysis of urine from patients undergoing CBP surgery

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