Dataset Information

Targeted analysis of PSMA in prostate cancer patient urinary extracellular vesicles labeled for PSMA and separated with asymmetrical flow field-flow fractionation coupled to a fluorescent light detector

ABSTRACT: Urine of 5 prostate cancer patients was collected, and labeled for PSMA. Targeted proteomic analysis was performed on the asymmetrical flow field-flow fractionated samples.

ORGANISM(S): Homo Sapiens

SUBMITTER: Pekka Rappu

PROVIDER: PXD031292 | panorama | Tue Mar 10 00:00:00 GMT 2026

REPOSITORIES: PanoramaPublic

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Publications

A One-Step Workflow for Size-Based Separation of Extracellular Vesicles With Integrated Surface Marker Detection.

Lippens Lien L Guilbert Niké N Van Dorpe Sofie S Deville Sarah S Boiy Robin R Roux Quentin Q Lumen Nicolaas N Miinalainen Ilkka I Rappu Pekka P Vandecasteele Katrien K Denys Hannelore H De Geest Bruno B De Wever Olivier O Hendrix An A

Journal of extracellular biology 20260303 3

Extracellular vesicles (EVs) are released by diverse cell types in biofluids and are increasingly studied in liquid biopsies as diagnostic and prognostic biomarkers for various diseases, including cancer. Typically, the analysis of EV-associated biomarker characteristics such as size and surface markers requires pre-purification from large volumes of complex biofluids, leading to longer turnaround times and more technical variability. To overcome these limitations, we developed a one-step workfl ...[more]

PMID: 41788818

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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.

Project description:The enteric bacterium Proteus mirabilis is a common cause of complicated urinary tract infections. In the study, microrarrays were used to analyze P. mirabilis gene expression in vivo from experimentally infected mice. Urine was collected at 1, 3, and 7d postinfection, and RNA was isolated from bacteria in the urine for transcriptional analysis. Across 9 microarrays, 471 genes were upregulated and 82 were downregulated in vivo compared to in vitro broth culture. Genes upregulated in vivo encoded MR/P fimbriae, urease, iron uptake systems, amino acid and peptide transporters, pyruvate metabolism, and portions of the TCA cycle. Flagella were downregulated. Ammonia assimilation gene glnA (glutamine synthetase) was repressed in vivo while gdhA (glutamate dehydrogenase) was upregulated in vivo. Contrary to our expectations, ammonia availability due to urease activity in P. mirabilis did not drive this gene expression. A gdhA mutant was growth-deficient in minimal medium with citrate as the sole carbon source, and loss of gdhA resulted in a significant fitness defect in the mouse model of urinary tract infection. Unlike Escherichia coli, which represses gdhA and upregulates glnA in vivo and cannot utilize citrate, the data suggest that P. mirabilis uses glutamate dehydrogenase to monitor carbon-nitrogen balance, and this ability contributes to the pathogenic potential of P. mirabilis in the urinary tract. Voided urine from female CBA/J mice infected with Proteus mirabilis was collected and pooled in RNA stabilizing reagent (RNAprotect). Urine was collected at 1, 3, and 7 d postinfection. RNA was isolated from urine and log-phase LB cultures, converted to cDNA, and labeled with CyDye. Three arrays were completed per time point (9 arrays total). Slides were scanned with a ScanArray Express microarray scanner (Perkin Elmer) at 10 μm resolution and quantified using ScanArray Express software. Resulting data were normalized by total intensity and median spot intensities were identified using MIDAS (v. 2.22) software.

Dataset Information

Targeted analysis of PSMA in prostate cancer patient urinary extracellular vesicles labeled for PSMA and separated with asymmetrical flow field-flow fractionation coupled to a fluorescent light detector

Publications

A One-Step Workflow for Size-Based Separation of Extracellular Vesicles With Integrated Surface Marker Detection.

Similar Datasets

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