Publication Date

2017

Document Type

Thesis

Committee Members

Nicholas DelRaso (Committee Member), John Paietta (Committee Member), Nicholas Reo (Advisor)

Degree Name

Master of Science (MS)

Abstract

Urinary metabolomics abilities in detection of biological effects (i.e., toxicity, disease) is compounded by high background variability. Previously, we showed mild kidney dysfunction was detectable under the stress imposed by furosemide (diuretic). Here we tested whether furosemide (FUR) can enhance the sensitivity to detect autosomal dominant polycystic kidney disease (ADPKD) at an early stage. ADPKD is one of the most common inherited renal disorders and is characterized by the growth of numerous cysts in the kidneys. Urinary metabolomics analyses, using nuclear magnetic resonance (NMR) spectroscopy, were conducted in control (WT) and diseased mice (RC) at 7 and 24 weeks of age. Urine samples were collected before (baseline) and after administration of furosemide or vehicle (saline). At 7 wks of age, the WT and RC mice showed different metabolite profiles before injection with FUR using Principle Components analysis (PCA) and OPLS discriminant analysis of the NMR data. We found [alpha] ketoglutarate, citrate, acetate, taurine, succinate and other metabolites that appear to be significant features that were responsible for the separation. There are multiple urinary NMR signals of diseased mice that have higher intensities than control mice, such as [alpha] ketoglutarate, citrate, acetate, taurine, succinate and other unknown metabolites. Significant increases in kidney weight per body weight ratio and cystic index in RC mice at 7 wks (D. Wallace, U Kansas) confirmed that disease progression was rapid. Thus, FUR could not be used to produce subtle differences in urinary metabolite profiles between WT and RC mice at 7 wks. At 24 wks of age, the WT and RC mice showed very similar urinary metabolite profiles at baseline (pre-dose), and preliminary data suggest that FUR helps to separate these groups. The "n" value of mouse urine samples at 24 wks was too small to test statistical significance of the work. ANOVA shows that some metabolites ([alpha] ketoglutarate/TSP and citrate/TSP) that are nearly significant (p=0.09). If n-value was larger, then these would likely be different. To confirm the hypothesis of this study, the study should be repeated using a larger sample size, and an animal model that develops cysts gradually, in order to show that furosemide has the potential to detect PKD at an early stage.

Page Count

78

Department or Program

Department of Biochemistry and Molecular Biology

Year Degree Awarded

2017

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