Project description:To investigate the transcriptome differences between wild-type and Vhl mutant mice in kidney stones process, we established CaOx-induced nephrolithiasis mouse model and performed RNA-sequencing.
Project description:Cystinuria is a rare renal genetic disease caused by mutations in cystine transporter genes and characterized by defective cystine reabsorption leading to kidney stones. In 14% of cases patients undergo nephrectomy, but given the difficulty to predict the evolution of the disease, the identification of markers of kidney damage would improve the follow up of patients with a higher risk. The aim of the present study is to develop a robust, reproducible and non-invasive methodology for proteomic analysis of urinary exosomes using high resolution mass spectrometry. A clinical pilot study, conducted on 8 cystinuria patients vs. 10 controls, highlighted 165 proteins, of which 38 were up-regulated, that separate cystinuria patients from controls, and further discriminate between severe and moderate forms of the disease. These proteins include markers of kidney injury, circulating proteins and a neutrophil signature. Analysis of selected proteins by immunobloting, performed on six additional cystinuria patients, validated the mass spectrometry data. To our knowledge, this is the first successful proteomic study in cystinuria unmasking potential role of inflammation in this disease. The workflow we have developed is applicable to investigate urinanry exosomes in different renal diseases and to search for diagnostic/prognostic markers.
Project description:The proposed project focuses on the study of urinary extracellular vesicles as a tool for the diagnosis and monitoring of kidney diseases, especially in patients undergoing kidney transplantation. We found a differential protein profile in patients with CKD before and after the transplant procedure.
Project description:Microarray analysis was used to assess the expression levels of lncRNAs and mRNAs in three pairs of Randall’s plaques tissues of CaOx renal stones patients and normal renal papillary tissues.Randall’s Plaques tissues of CaOx renal stones patients were obtained from patients with idiopathic CaOx renal stone who underwent percutaneous nephrolithotomy. Normal renal papillary tissues were obtained from patients with renal tumor who underwent nephrectomy, the tissues were obtained from papillary without tumor invasion. Randall’s Plaques tissues of CaOx renal stones patients were obtained from patients with idiopathic CaOx renal stone who underwent percutaneous nephrolithotomy. Normal renal papillary tissues were obtained from patients with renal tumor who underwent nephrectomy, the tissues were obtained from papillary without tumor invasion.
Project description:Abnormal N-glycosylation of uromodulin (Umod, also known as Tamm-Horsfall protein) has been implicated in the pathogenesis of various urinary system diseases. However, the site-specific N-glycosylation patterns of urinary Umod in healthy controls (HCs) versus those with kidney stones (KSs) have yet to be detailed. In this study, we isolated and purified Umod from the urine of 24 patients with calcium oxalate (CaOx) KSs and 24 HCs using a refined diatomaceous earth adsorption method. After digestion with trypsin and Glu-C, intact N-glycopeptides (IGPs) were enriched via hydrophilic interaction liquid chromatography (HILIC) and analyzed via EThcD-sceHCD-MS/MS. Data processing was conducted using Byonic and PANDA software. A total of 700 IGPs, 8 N-glycosites, and 145 N-glycans were identified, marking the most extensive identification of N-glycosylation in Umod to date. Through quantitative and site-specific N-glycosylation analyses, the study found 39 IGPs to be up-regulated (p<0.05, FC>1.5) and 60 down-regulated IGPs (p<0.05, FC<0.67) in KS groups. Additionally, the analysis of N-glycan composition and site-specific N-glycosylation highlighted KS-specific N-glycosylation modifications. Overall, these results suggest that variations in Umod N-glycosylation may critically contribute to the formation of CaOx kidney stones by modulating its function.