Project description:Transcriptome analysis of growth hormone dependant genes in glomerular podocytes Differentiated human glomerular podocytes in culture exposed to growth hormone for 0 min, 2 min, 5 min, 15 min, and 30 min. Total RNA is extracted and subjected to microarray analysis.
Project description:The growth hormone plays a significant role in normal renal function and overactive growth hormone signaling has been implicated in proteinuria in diabetes. Earlier studies from our group have shown that the glomerular podocytes, which play an essential role in renal filtration, express the growth hormone receptor, suggesting the direct action of growth hormone on these cells. Nevertheless, the precise mechanism and the downstream pathways that are induced by the excess growth hormone in these podocytes leading to diabetic nephropathy are not clearly established. To compressively understand the growth hormone’s effect on podocytes at transcript level we performed RNA-Sequencing. Conditionally immortalized human podocytes were employed in this study.
Project description:Purpose: Next-generation sequencing (NGS) was used to define the transcriptome of native mouse podocytes and non-podocytes glomerular cells as part of a project aiming to define the molecular fingerprint of mouse podocytes. Method: Glomeruli from 29 Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo/J x hNPHS2Cre mice at the age of 10 weeks were purified and a single cell solution was prepared to seperate GFP-expressing (podocytes) and GFP-negative (non-podocytes glomerular cells) cells by FACS sorting. RNA was extracted and prepared for further analysis using directional, polyA+ library preparation. An Illumina HiSeq2500 was used for a paired-end sequencing of 100 cycles . Salmon and Sleuth were used for downstream analysis. Results: A total of 100 Million reads each from podocytes and non-podocytes glomerular cells could be used for further analysis.
Project description:Calcimimetics such as R568 are renoprotective and the underlying mechanism is not yet fully clarified. We consider the importance of podocytes in mediating the beneficial effect of calcimimetics on kidney. Affymetrix array type Moe430_2 was used to study the effect of R-568. Analysis of differential gene expression indicates that R-568 acts antioxidative, stabilizes cytoskeleton, enhances cell cycle control and suppresses apoptosis in podocytes. This suggests that calcimimetics limit podocyte damage by antiapoptotic and cytoskeleton stabilizing effects and may thus constitute a new pharmacological approach in the prevention and treatment of glomerular disease. Podocytes treated with and without R568 were used for RNA extraction and hybridization on Affymetrix microarrays.
Project description:We profiled the somatic landscape of 21 growth hormone (GH) -secreting pituitary adenomas using somatic copy-number alteration (SCNA), whole-genome sequencing (WGS), bisulfate sequencing, and transcriptome approaches. See details in Valimaki et al. Genetic and epigenetic characterization of growth hormone (GH) - secreting pituitary tumors. Manuscript in preparation, 2019.
Project description:Calcimimetics such as R568 are renoprotective and the underlying mechanism is not yet fully clarified. We consider the importance of podocytes in mediating the beneficial effect of calcimimetics on kidney. Affymetrix array type Moe430_2 was used to study the effect of R-568. Analysis of differential gene expression indicates that R-568 acts antioxidative, stabilizes cytoskeleton, enhances cell cycle control and suppresses apoptosis in podocytes. This suggests that calcimimetics limit podocyte damage by antiapoptotic and cytoskeleton stabilizing effects and may thus constitute a new pharmacological approach in the prevention and treatment of glomerular disease.
Project description:Podocytes, highly differentiated glomerular epithelial cells, are essential for the maintenance of glomerular filtration barrier. Podocyte dysfunction in podocytes is a major determinant of proteinuric kidney disease. By RNA sequencing analysis in ADR-treated podocytes with or without MYDGF overexpression, we observed the significant changes of genes important in regulating cell cycle in podocytes with ADR treatment.
Project description:The effect of miRNA delivery from glomerular endothelial cells (GEnCs) to podocytes in vitro was examined by miRNA epression in podocytes untreated and treated with extracellular vesicles from GEnCs under varied activation conditions.
Project description:The effect of miRNA delivery from glomerular endothelial cells (GEnCs) to podocytes in vitro was examined by mRNA epression in podocytes untreated and treated with extracellular vesicles from GEnCs under varied activation conditions.