Project description:Glucocorticoid resistance complicates the treatment of ~20% of children with nephrotic syndrome, yet the molecular basis for resistance remains unclear. We generated the transcriptome profile by RNA sequencing of peripheral blood leukocytes from children obtained both at initial nephrotic syndrome presentation and after ~7 weeks of glucocorticoid therapy to identify genes or a gene panel able to differentiate steroid sensitive from steroid resistant nephrotic syndrome. RNA -seq analysis was followed by in-silico algorithm-based approaches and subsequent biochemical analyses on relevant candidate gene with important roles in podocyte and glomerular pathophysiology, using both patient samples and experimental models of nephrotic syndrome and podocyte injury.
Project description:This study aimed to evaluate gene expression patterns in urinary sediment samples of children with steroid-resistant nephrotic syndrome (SRNS).
Project description:We have perfomed Visium HD spatial transcriptomics on archival FFPE kidney biopsies from patients with different manifistations of nephrotic syndrome, i.e., steriod sensitive, frequent relapsing and/or steroid dependent, and focal segmental glomerulosclerosis. Using a pre-trained deep learning model from StarDist to segment nuclei, we reconstructed single-nucleus level spatial trancriptomic datasets through custom binning of subcellular capture areas. We integrated the datasets from the different patients and indentified conserved cell types. Moreover, we identified patient specific transcriptomic differences in the podocyte cell population. This article demonstrates the usefullness of this approach to future studies of pathophysilogical mechanisms in idiopathic nephrotic syndrome.
Project description:Nephrotic syndrome (NS) is an unfavorable disease with heterogeneous causes and variable prognosis. One of the clinically crucial prognostic feature is the response to initial glucocorticoid treatment. Whereas in some children with steroid-sensitive nephrotic syndrome (SSNS) the treatment may induce long-term remission of the disease, steroid-resistant cases (SRNS) are at risk of renal failure requiring renal replacement therapy. The aim of this project was to explore whether sera from children with the two main clinically defined types of NS induce changes in transcriptome of in-vitro cultures of immortalized human podocytes. After 3 days of culture, total RNA was isolated and then library from polyadenylated RNA was prepared and RNA sequencing was performed. Log2 (fold-changes) were calculated to describe differential gene expression. Within the Reactome online tool, Camera methodology was implemented to identify functionally linked gene groups (pathways) that could distinguish the two patient groups. Significant transcriptome differences were found between samples from children with SSNS and SRNS. This may help to reveal the molecular mechanisms of the disease and open up for an effective individualized treatment.
2022-10-14 | GSE215231 | GEO
Project description:A gut microbiota study about steroid children steroid-dependent primary nephrotic syndrome
