Confounding Factors Impacting microRNA Expression in Human Saliva: Methodological and Biological Considerations

There is growing interest in saliva microRNAs (miRNAs) as non-invasive biomarkers for human disease. Such an approach requires understanding how differences in experimental design affect miRNA expression. Variations in technical methodologies, coupled with inter-individual variability may reduce stu...

Full description

Saved in:
Bibliographic Details
Published in:Genes 2022-10, Vol.13 (10), p.1874
Main Authors: Sullivan, Rhea, Montgomery, Austin, Scipioni, Anna, Jhaveri, Pooja, Schmidt, Adam T, Hicks, Steven D
Format: Article
Language:English
Subjects:
Analysis
Biological markers
Biomarkers
Biomarkers - metabolism
Body mass index
Cohort analysis
Confounding (Statistics)
Discriminant analysis
Gene expression
Humans
MicroRNA
MicroRNAs
MicroRNAs - genetics
MicroRNAs - metabolism
miRNA
Physiological aspects
Reproducibility of Results
Saliva
Saliva - chemistry
Salivary glands
secretions
Statistical analysis
Traumatic brain injury
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:There is growing interest in saliva microRNAs (miRNAs) as non-invasive biomarkers for human disease. Such an approach requires understanding how differences in experimental design affect miRNA expression. Variations in technical methodologies, coupled with inter-individual variability may reduce study reproducibility and generalizability. Another barrier facing salivary miRNA biomarker research is a lack of recognized "control miRNAs". In one of the largest studies of human salivary miRNA to date (922 healthy individuals), we utilized 1225 saliva samples to quantify variability in miRNA expression resulting from aligner selection (Bowtie1 vs. Bowtie2), saliva collection method (expectorated vs. swabbed), RNA stabilizer (presence vs. absence), and individual biological factors (sex, age, body mass index, exercise, caloric intake). Differential expression analyses revealed that absence of RNA stabilizer introduced the greatest variability, followed by differences in methods of collection and aligner. Biological factors generally affected a smaller number of miRNAs. We also reported coefficients of variations for 643 miRNAs consistently present in saliva, highlighting several salivary miRNAs to serve as reference genes. Thus, the results of this analysis can be used by researchers to optimize parameters of salivary miRNA measurement, exclude miRNAs confounded by numerous biologic factors, and identify appropriate miRNA controls.
ISSN:2073-4425
2073-4425
DOI:10.3390/genes13101874