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...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-10-01
|
| Series: | Genes |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4425/13/10/1874 |
| _version_ | 1797407207826915328 |
|---|---|
| author | Rhea Sullivan Austin Montgomery Anna Scipioni Pooja Jhaveri Adam T. Schmidt Steven D. Hicks |
| author_facet | Rhea Sullivan Austin Montgomery Anna Scipioni Pooja Jhaveri Adam T. Schmidt Steven D. Hicks |
| author_sort | Rhea Sullivan |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-09T03:37:58Z |
| format | Article |
| id | doaj.art-e63c0c79d4e145e8bafe7510540de099 |
| institution | Directory Open Access Journal |
| issn | 2073-4425 |
| language | English |
| last_indexed | 2024-03-09T03:37:58Z |
| publishDate | 2022-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Genes |
| spelling | doaj.art-e63c0c79d4e145e8bafe7510540de0992023-12-03T14:45:30ZengMDPI AGGenes2073-44252022-10-011310187410.3390/genes13101874Confounding Factors Impacting microRNA Expression in Human Saliva: Methodological and Biological ConsiderationsRhea Sullivan0Austin Montgomery1Anna Scipioni2Pooja Jhaveri3Adam T. Schmidt4Steven D. Hicks5Department of Pediatrics, Penn State Hershey College of Medicine, Hershey, PA 17033, USADepartment of Pediatrics, Penn State Hershey College of Medicine, Hershey, PA 17033, USADepartment of Pediatrics, Penn State Hershey College of Medicine, Hershey, PA 17033, USADepartment of Pediatrics, Penn State Hershey College of Medicine, Hershey, PA 17033, USADepartment of Psychological Sciences, Texas Tech University, Lubbock, TX 79409, USADepartment of Pediatrics, Penn State Hershey College of Medicine, Hershey, PA 17033, USAThere 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.https://www.mdpi.com/2073-4425/13/10/1874miRNAsalivarybiomarkerhousekeepingstabilizer |
| spellingShingle | Rhea Sullivan Austin Montgomery Anna Scipioni Pooja Jhaveri Adam T. Schmidt Steven D. Hicks Confounding Factors Impacting microRNA Expression in Human Saliva: Methodological and Biological Considerations Genes miRNA salivary biomarker housekeeping stabilizer |
| title | Confounding Factors Impacting microRNA Expression in Human Saliva: Methodological and Biological Considerations |
| title_full | Confounding Factors Impacting microRNA Expression in Human Saliva: Methodological and Biological Considerations |
| title_fullStr | Confounding Factors Impacting microRNA Expression in Human Saliva: Methodological and Biological Considerations |
| title_full_unstemmed | Confounding Factors Impacting microRNA Expression in Human Saliva: Methodological and Biological Considerations |
| title_short | Confounding Factors Impacting microRNA Expression in Human Saliva: Methodological and Biological Considerations |
| title_sort | confounding factors impacting microrna expression in human saliva methodological and biological considerations |
| topic | miRNA salivary biomarker housekeeping stabilizer |
| url | https://www.mdpi.com/2073-4425/13/10/1874 |
| work_keys_str_mv | AT rheasullivan confoundingfactorsimpactingmicrornaexpressioninhumansalivamethodologicalandbiologicalconsiderations AT austinmontgomery confoundingfactorsimpactingmicrornaexpressioninhumansalivamethodologicalandbiologicalconsiderations AT annascipioni confoundingfactorsimpactingmicrornaexpressioninhumansalivamethodologicalandbiologicalconsiderations AT poojajhaveri confoundingfactorsimpactingmicrornaexpressioninhumansalivamethodologicalandbiologicalconsiderations AT adamtschmidt confoundingfactorsimpactingmicrornaexpressioninhumansalivamethodologicalandbiologicalconsiderations AT stevendhicks confoundingfactorsimpactingmicrornaexpressioninhumansalivamethodologicalandbiologicalconsiderations |