Characterization of miRNA-regulated networks, hubs of signaling, and biomarkers in obstruction-induced bladder dysfunction
Ali Hashemi Gheinani, … , Fiona C. Burkhard, Katia Monastyrskaya
Ali Hashemi Gheinani, … , Fiona C. Burkhard, Katia Monastyrskaya
Published January 26, 2017
Citation Information: JCI Insight. 2017;2(2):e89560. https://doi.org/10.1172/jci.insight.89560.
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Research Article Aging Muscle biology

Characterization of miRNA-regulated networks, hubs of signaling, and biomarkers in obstruction-induced bladder dysfunction

Abstract

Bladder outlet obstruction (BOO) induces significant organ remodeling, leading to lower urinary tract symptoms accompanied by urodynamic changes in bladder function. Here, we report mRNA and miRNA transcriptome sequencing of bladder samples from human patients with different urodynamically defined states of BOO. Patients’ miRNA and mRNA expression profiles correlated with urodynamic findings. Validation of RNA sequencing results in an independent patient cohort identified combinations of 3 mRNAs (NRXN3, BMP7, UPK1A) and 3 miRNAs (miR-103a-3p, miR-10a-5p, miR-199a-3p) sufficient to discriminate between bladder functional states. All BOO patients shared cytokine and immune response pathways, TGF-β and NO signaling pathways, and hypertrophic PI3K/AKT signaling pathways. AP-1 and NFkB were dominant transcription factors, and TNF-α was the top upstream regulator. Integrated miRNA-mRNA expression analysis identified pathways and molecules targeted by differentially expressed miRNAs. Molecular changes in BOO suggest an increasing involvement of miRNAs in the control of bladder function from the overactive to underactive/acontractile states.

Authors

Ali Hashemi Gheinani, Bernhard Kiss, Felix Moltzahn, Irene Keller, Rémy Bruggmann, Hubert Rehrauer, Catharine Aquino Fournier, Fiona C. Burkhard, Katia Monastyrskaya

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Figure 5

QPCR validation of NGS data and 3D scatter plots of 3-mRNA and 3-miRNA signatures of BOO states.

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QPCR validation of NGS data and 3D scatter plots of 3-mRNA and 3-miRNA s...
QPCR using miRNA and mRNA expression assays was carried out in controls and independent BOO-induced LUTD patient groups (n ≥ 12 samples per group). Genes and miRNAs, altered in all BOO patients as well as group-specific miRNAs and mRNAs (ROC analysis), were included. (A) Analysis of mRNA expression in BOO patients. Hierarchical clustering and heatmap of normalized Ct values of 11 candidate mRNAs (y axis) in 49 patients (x axis). (B) Analysis of miRNA expression in BOO patients. Hierarchical clustering and heatmap of normalized Ct values of 18 candidate miRNAs (y axis) in 28 patients (x axis). The clustering of patients based on QPCR results for 11 mRNAs and 18 miRNAs was similar to the clustering of patients based on NGS data. (C) Regulation of 6 significantly changed mRNAs in 49 patients. QPCR results in 3 patient groups are shown in a box-and-whisker plot as log2 fold changes compared with controls, representing minimum, first quartile, median, third quartile, and maximum. *P < 0.05, **P < 0.001 (ANOVA). (D) Regulation of 6 significantly changed miRNAs in 28 patients. QPCR results in 3 patient groups are shown in a box-and-whisker plot as log2 fold changes compared with controls. *P < 0.05, **P < 0.001 (ANOVA), representing minimum, first quartile, median, third quartile, and maximum. (E) 3D scatter plots and point identification for 3 mRNA biomarkers. Log2 fold change values of NRXN3, BMP7, and UPK1A mRNAs in bladder biopsies of 49 patients with different BOO states are plotted. Concentration ellipsoids are drawn to show the DO patients in green, BO patients in blue, and UA patients in orange. (F) 3D scatter plots and point identification for 3 miRNA biomarkers. Log2 fold change values of miRNAs miR-103a-3p, hsa-miR10a-5p, and hsa-miR-199a-3p suggested by ROC analysis in bladder biopsies of 18 patients with different states of BOO are plotted. The 3-mRNA and 3-miRNA signatures are sufficient to discriminate DO, BO, and UA groups from each other. C, controls; DO, obstructed patients with detrusor overactivity; BO, obstructed patients without detrusor overactivity; UA, obstructed patients with underactive bladders.