Optimization of Urinary Extracellular Vesicles Analysis to Study Kidney Disease in People with Cystic Fibrosis

Background: People with Cystic Fibrosis (PwCF) have higher risk of kidney disease, but there are no markers predicting the risk.
Urinary extracellular vesicles (uEVs) are nanoparticles and reliable sources of potential biomarkers of diseases. We hypothesize that specific uEV markers may identify PwCF at risk of kidney injury.
Methods: We isolated uEVs from PwCF and healthy controls (HCs). Differential centrifugation was used to enrich uEVs. Nanoparticle tracking analysis and Electron microscopy were used to examine the size, concentration, and morphology of uEVs. Proteomics approach and pathway analyses were used to detect uEV markers and enriched pathways.
Results: We identified 1404 proteins including uEV markers of the urinary tract from HCs and achieved high reproducibility of protein coverage.
Conclusions: We will compare the uEVs’ proteome to identify reduced kidney function signatures in PwCF. The proteomic signatures will be validated to identify their predictive ability for kidney injury before standard clinical tests.