Biomarkers of nucleic acid oxidation - A summary state-of-the-art
Mu-Rong Chao1, Mark D Evans2, Chiung-Wen Hu3
1Department of Occupational Safety and Health, Chung Shan Medical University, Taichung, 402, Taiwan; Department of Occupational Medicine, Chung Shan Medical University Hospital, Taichung, 402, Taiwan.
Abstract:
Oxidatively generated damage to DNA has been implicated in the pathogenesis of a wide variety of diseases. Increasingly, interest is also focusing upon the effects of damage to the other nucleic acids, RNA and the (2'-deoxy-)ribonucleotide pools, and evidence is growing that these too may have an important role in disease. LC-MS/MS has the ability to provide absolute quantification of specific biomarkers, such as 8-oxo-7,8-dihydro-2'-deoxyGuo (8-oxodG), in both nuclear and mitochondrial DNA, and 8-oxoGuo in RNA. However, significant quantities of tissue are needed, limiting its use in human biomonitoring studies. In contrast, the comet assay requires much less material, and as little as 5 μL of blood may be used, offering a minimally invasive means of assessing oxidative stress in vivo, but this is restricted to nuclear DNA damage only. Urine is an ideal matrix in which to non-invasively study nucleic acid-derived biomarkers of oxidative stress, and considerable progress has been made towards robustly validating these measurements, not least through the efforts of the European Standards Committee on Urinary (DNA) Lesion Analysis. For urine, LC-MS/MS is considered the gold standard approach, and although there have been improvements to the ELISA methodology, this is largely limited to 8-oxodG. Emerging DNA adductomics approaches, which either comprehensively assess the totality of adducts in DNA, or map DNA damage across the nuclear and mitochondrial genomes, offer the potential to considerably advance our understanding of the mechanistic role of oxidatively damaged nucleic acids in disease.
Insights
Oxidative damage to DNA, RNA, and nucleotide pools contributes to disease. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) and urine analysis offer promising methods for detecting these biomarkers.
Area of Science:
- Biochemistry
- Molecular Biology
- Disease Pathogenesis
Background:
- Oxidative damage to nucleic acids (DNA, RNA) and nucleotide pools is increasingly linked to various diseases.
- Understanding these damages is crucial for disease mechanism elucidation and biomarker development.
Purpose of the Study:
- To review methods for assessing oxidative damage to nucleic acids and their relevance in disease.
- To highlight the potential of urine-based biomarkers for non-invasive oxidative stress monitoring.
Main Methods:
- Liquid chromatography-tandem mass spectrometry (LC-MS/MS) for absolute quantification of biomarkers like 8-oxo-7,8-dihydro-2'-deoxyGuo (8-oxodG) and 8-oxoGuo.
- Comet assay for assessing nuclear DNA damage in small sample volumes.
- Urine analysis as a non-invasive matrix for biomarker detection, with LC-MS/MS as the gold standard.
Main Results:
- LC-MS/MS provides precise quantification but requires significant tissue, limiting its use in human biomonitoring.
- The comet assay is minimally invasive but restricted to nuclear DNA damage.
- Urine analysis shows promise for non-invasive oxidative stress assessment, with validated measurement approaches.
Conclusions:
- Oxidative damage to nucleic acids plays a significant role in disease pathogenesis.
- Urine-based biomarkers, analyzed by LC-MS/MS, offer a viable non-invasive approach for assessing oxidative stress.
- Emerging DNA adductomics techniques hold potential for advancing mechanistic understanding of oxidative nucleic acid damage in disease.
