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Identification of Radiation-Induced miRNA Biomarkers Using the CGL1 Cell Model System
Jayden Peterson1, Christopher D McTiernan2, Christopher Thome1,2,3,4
1School of Natural Sciences, Laurentian University, Sudbury, ON P3E 2C6, Canada.
Abstract:
MicroRNAs (miRNAs) have emerged as a potential class of biomolecules for diagnostic biomarker applications. miRNAs are small non-coding RNA molecules, produced and released by cells in response to various stimuli, that demonstrate remarkable stability in a wide range of biological fluids, in extreme pH fluctuations, and after multiple freeze-thaw cycles. Given these advantages, identification of miRNA-based biomarkers for radiation exposures can contribute to the development of reliable biological dosimetry methods, especially for low-dose radiation (LDR) exposures. In this study, an miRNAome next-generation sequencing (NGS) approach was utilized to identify novel radiation-induced miRNA gene changes within the CGL1 human cell line. Here, irradiations of 10, 100, and 1000 mGy were performed and the samples were collected 1, 6, and 24 h post-irradiation. Corroboration of the miRNAome results with RT-qPCR verification confirmed the identification of numerous radiation-induced miRNA expression changes at all doses assessed. Further evaluation of select radiation-induced miRNAs, including miR-1228-3p and miR-758-5p, as well as their downstream mRNA targets, Ube2d2, Ppp2r2d, and Id2, demonstrated significantly dysregulated reciprocal expression patterns. Further evaluation is needed to determine whether the candidate miRNA biomarkers identified in this study can serve as suitable targets for radiation biodosimetry applications.
Insights
MicroRNAs (miRNAs) show promise as radiation exposure biomarkers due to their stability. This study identified radiation-induced miRNA changes using next-generation sequencing, supporting their use in biological dosimetry.
Area of Science:
- Biomolecular research
- Molecular biology
- Genomics
Background:
- MicroRNAs (miRNAs) are small, stable non-coding RNA molecules released by cells.
- Their stability in biological fluids and resistance to environmental stress make them ideal biomarker candidates.
- Accurate biological dosimetry for radiation exposure is crucial, especially for low-dose radiation (LDR).
Purpose of the Study:
- To identify novel radiation-induced miRNA expression changes using an miRNAome next-generation sequencing (NGS) approach.
- To evaluate the potential of identified miRNAs as biomarkers for radiation biodosimetry.
- To investigate the reciprocal expression patterns of candidate miRNAs and their mRNA targets.
Main Methods:
- Utilized miRNAome next-generation sequencing (NGS) on the CGL1 human cell line.
- Exposed cells to 10, 100, and 1000 mGy radiation doses.
- Collected samples at 1, 6, and 24 hours post-irradiation.
- Verified miRNA expression changes using RT-qPCR.
- Analyzed downstream mRNA targets of selected miRNAs.
Main Results:
- Identified numerous radiation-induced miRNA expression changes across all tested doses and time points.
- Confirmed radiation-induced changes through RT-qPCR validation.
- Observed significantly dysregulated reciprocal expression patterns between specific miRNAs (e.g., miR-1228-3p, miR-758-5p) and their mRNA targets (e.g., Ube2d2, Ppp2r2d, Id2).
Conclusions:
- The study successfully identified radiation-induced miRNA expression changes in human cells.
- Candidate miRNAs, such as miR-1228-3p and miR-758-5p, show potential as biomarkers for radiation biodosimetry.
- Further research is required to validate these miRNA candidates for practical biodosimetry applications.
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