Identification of miR-16 as an endogenous reference gene for the normalization of urinary exosomal miRNA expression data from CKD patients

Chronic kidney disease (CKD) is a severe disorder with an increasing incidence worldwide. An early detection may help to prevent its progression and to minimize the risk of cardiovascular diseases as one of the major comorbidities. Recently, extracellular miRNAs like urinary exosomal miRNAs became o...

Full description

Saved in:
Bibliographic Details
Published in:PloS one 2017-08, Vol.12 (8), p.e0183435-e0183435
Main Authors: Lange, Tim, Stracke, Sylvia, Rettig, Rainer, Lendeckel, Uwe, Kuhn, Jana, Schlüter, Rabea, Rippe, Volkhard, Endlich, Karlhans, Endlich, Nicole
Format: Article
Language:English
Subjects:
Aged
Analysis
Bioindicators
Biology
Biology and Life Sciences
Biomarkers
Biomarkers - urine
Body fluids
Cancer
Cardiovascular diseases
Chronic kidney failure
Complications and side effects
Data processing
Development and progression
Diabetes
Exosomes
Exosomes - metabolism
Female
Gene expression
Gene Expression Regulation - genetics
Health care
Health risks
Heart diseases
Humans
Incidence
Kidney diseases
Kidney transplantation
Male
Medicine
Medicine and Health Sciences
MicroRNA
MicroRNAs
MicroRNAs - urine
Microscopy
Middle Aged
miRNA
Patients
Prevention
Reference Standards
Renal Insufficiency, Chronic - pathology
Renal Insufficiency, Chronic - urine
Research and analysis methods
Ribonucleic acid
Risk factors
Risk reduction
RNA
snRNA
Studies
Transplants & implants
Urine
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Chronic kidney disease (CKD) is a severe disorder with an increasing incidence worldwide. An early detection may help to prevent its progression and to minimize the risk of cardiovascular diseases as one of the major comorbidities. Recently, extracellular miRNAs like urinary exosomal miRNAs became of great interest as non-invasive biomarkers which can be determined by RT-qPCR. But until now, there is no consensus regarding the normalization of miRNAs isolated from body fluids. The present study analyzed the miRNAs miR-16, miR-92a, miR-21, miR-124a and the small nuclear RNA RNU6B for their applicability as an endogenous reference gene in expression studies of exosomal miRNAs isolated from CKD patients. For this purpose, miRNA expression levels were determined by RT-qPCR after the isolation of urinary exosomes from 33 CKD patients and from 5 healthy controls. Expression data was analyzed with the normalization determination software NormFinder, BestKeeper, GeNorm and DeltaCt. Our results revealed an abundant expression of the four candidate miRNAs in urinary exosomes and no detectable expression of RNU6B. We identified miR-16 as the most stable endogenous reference gene in our data set, making it a suitable endogenous reference gene for miRNA studies of urinary exosomes derived from CKD patients.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0183435