Browsing by Subject "Aldosterone"

Now showing 1 - 3 of 3

Open Access
Development of a shiny application for comparative transcriptomics and differential gene expression analysis
(2022-09) Leka, Ronaldo
RNA sequencing has proven to be an effective technique for divulging information about the transcriptome in molecular biology research. Compared to microarrays and early methods of cDNA sequencing, high-throughput RNA sequencing has better resolution, low background noise, a higher range to quantify gene expression, and relatively lower cost. The development of sequencing technique has led to the development of tools for analyzing the high volume of data that is generated. Statistical methods for normalizing, filtering, performing exploratory and differential analysis, and other functional analyses based on RNA sequencing count data have made RNA sequencing one of the most popular techniques in genomics. To help facilitate the use of such statistical tools, web applications developed in R using the shiny package offer an advantageous environment where researchers can use a graphical interface to give inputs and instructions to the underlying server-side libraries that analyze and generate results in tables and plots. This thesis presents a new tool that has been developed for exploratory analysis, data normalization and filtering, differential gene expression analysis (DGEA), correlation analysis, principal component analysis, and functional analysis such as over-representation analysis and gene set enrichment analysis. When compared to other available applications, this new application offers the ability to run multiple methods for DGEA and compare results between them, leading to the output of gene sets that are discovered as DEGs in multiple tests. Here I present the features of this application in detail where I aim to improve upon the applications that are available in the literature. An example dataset from our lab was also investigated by this RNA-seq tool leading to a better understanding of Mineralocorticoid Receptor (MR) signaling in breast cancer.
Open Access
Increased SGK1 activity potentiates mineralocorticoid/NaCl-induced kidney injury
(American Physiological Society, 2021-04-08) Sierra-Ramos, Catalina; Velazquez-Garcia, Silvia; Keskus, Ayşe Gökçe; Vastola-Mascolo, Arianna; Rodríguez-Rodríguez, Ana E.; Luis-Lima, Sergio; Hernández, Guadalberto; Navarro-González, Juan F.; Porrini, Esteban; Konu, Özlen; Alvarez de la Rosa, Diego
Serum and glucocorticoid-regulated kinase 1 (SGK1) stimulates aldosterone-dependent renal Na reabsorption and modulates blood pressure. In addition, genetic ablation or pharmacological inhibition of SGK1 limits the development of kidney inflammation and fibrosis in response to excess mineralocorticoid signaling. In this work, we tested the hypothesis that a systemic increase in SGK1 activity would potentiate mineralocorticoid/salt-induced hypertension and kidney injury. To that end, we used a transgenic mouse model with increased SGK1 activity. Mineralocorticoid/salt-induced hypertension and kidney damage was induced by unilateral nephrectomy and treatment with deoxycorticosterone acetate and NaCl in the drinking water for 6 wk. Our results show that although SGK1 activation did not induce significantly higher blood pressure, it produced a mild increase in glomerular filtration rate, increased albuminuria, and exacerbated glomerular hypertrophy and fibrosis. Transcriptomic analysis showed that extracellular matrix-and immune response-related terms were enriched in the downregulated and upregulated genes, respectively, in transgenic mice. In conclusion, we propose that systemically increased SGK1 activity is a risk factor for the development of mineralocorticoid-dependent kidney injury in the context of low renal mass and independently of blood pressure. NEW and NOTEWORTHY Increased activity of the protein kinase serum and glucocorticoid-regulated kinase 1 may be a risk factor for accelerated renal damage. Serum and glucocorticoid-regulated kinase 1 expression could be a marker for the rapid progression toward chronic kidney disease and a potential therapeutic target to slow down the process. © 2021 American Physiological Society. All rights reserved.
Open Access
Regulation of mineralocorticoid receptor and its downstream targets by estrogen and aldosterone in breast cancer
(2016-11) Çoban, Bircan
Many women suffer from breast cancer worldwide thus accurate diagnosis of this disease has become an important issue for treatment options and improved clinical outcomes. Members of steroid hormone receptors, are a subfamily of nuclear receptors can serve as biomarkers in molecular classification of breast cancer. One of these, Mineralocorticoid Receptor (MR) takes part in many physiological processes in epithelial tissues including mammary epithelia, yet it is not well studied in the context of breast cancer. In this thesis, we investigated expression patterns of MR together with Glucocorticoid Receptor (GR) across multiple breast cancer cell lines at the protein level. Our study revealed that expressions of MR and GR were modulated in breast cancer as a subtype specific manner. We then enquired regulation of MR and its downstream targets, SGK1, NEDD4-2 and subunits of ENaC i.e., α, β and γ, by estrogen (E2) and aldosterone (ALDO) treatment in breast cancer via qPCR and Western Blotting. We found differential responses in expression of MR and its downstream targets to E2 and ALDO suggesting ER status was an important mediator of MR action. We also overexpressed MR in MCF7 cells and then showed that MR, NEDD4-2, β and γENaC mRNA levels increased in response to ALDO only when MR was overexpressed.