Plasma microRNAs as Biomarkers for Predicting Radiotherapy Treatment-Induced Cardiotoxicity in Lung Cancer

Abstract

Background: Lung cancer is the second most common malignancy and stands as a leading cause of cancer-related deaths worldwide. Currently, one of the main treatment options for lung cancer is radiotherapy, but this treatment is associated with complications, such as an increased risk of cardiac-related morbidity and mortality. However, currently available methods for predicting radiation-induced heart disease (RIHD) remain suboptimal. Methods: In this pilot study, using the RT-qPCR method, we analyzed the expression levels of six miRNAs (miRNA-1-3p, miRNA-21-5p, miRNA-24-3p, miRNA-29a-3p, miRNA-34a-5p, and miRNA-222-3p). Results: Fourteen pairs of locally advanced non-small-cell lung cancer patients' plasma samples, taken before and after radiotherapy, were examined. It was observed that miRNA-1-3p, miRNA-21-5p, miRNA-24-3p, miRNA-29a-3p, and miRNA-222-3p were downregulated, while miRNA-34a-5p was upregulated in lung cancer patients' plasma after treatment. Additionally, after definitive radiotherapy, patients with an increased NT-proBNP value displayed a statistically significant difference in miRNA-222-3p levels compared to the normal range of this indicator. The panel of the combined four miRNAs for assessing the risk of cardiac comorbidities demonstrated an AUC of 0.79, sensitivity of 71.43%, and specificity of 100%, with further improved values upon integration with clinical biomarker NT-proBNP. Conclusions: This pilot study shows that the identification of changes in miRNA expression levels in lung cancer patients' plasma before and after radiotherapy could be used for the early diagnosis of RIHD.

Keywords: RIHD; microRNA; non-small-cell lung cancer.

Figure 1

Heatmap of miRNA expression fold change after treatment (n = 14). Gray cells indicate no data.

Figure 2

Heatmap of miRNA expression fold change after treatment among patients diagnosed with malignant neoplasm of the upper lobe, bronchus, or lung (n = 8) and those diagnosed with overlapping malignant neoplasm of the bronchus and lung (n = 4). The group diagnosed with malignant neoplasm of the middle lobe, bronchus, or lung is not shown due to its small sample size. Gray cells indicate no data. “*”—indicates a statistically significant value.

Figure 3

Correlation between miRNA expression fold change after therapy and regular (non-)use of cardiovascular drugs (n = 10). MiRNA-1-3p and miRNA-34a-3p are not shown due to missing data. Boxes indicate the ${log}_2$ normalized expression data, and the line inside each box represents the median. Whiskers denote the minimum and maximum values.

Figure 4

MiRNA expression fold change after radiotherapy in patients with an increase in natriuretic peptide concentration versus non-increase (norm) (n = 10). MiRNA-34a-3p is not shown due to missing data. Boxes indicate the ${log}_2$ normalized expression data, and the line inside each box represents the median. Whiskers denote the minimum and maximum values.

Figure 5

MiRNA expression fold change after radiotherapy in patients with cardiac comorbidities versus without (norm) (n = 13). MiRNA-1-3p and miRNA-34a-5p were excluded from the analysis due to missing data. Boxes indicate the ${log}_2$ normalized expression data, and the line inside each box represents the median. Whiskers denote the minimum and maximum values.

Figure 6

ROC curve analysis of the miRNA expression fold change in relation to cardiac comorbidities after radiotherapy (n = 13). MiRNA-1-3p and miRNA-34a-5p were excluded from the analysis due to missing data.

Figure 7

ROC curve analysis of miRNA expression fold change and NT-proBNP status in relation to cardiac comorbidities after radiotherapy (n = 10). MiRNA-1-3p and miRNA-34a-5p were excluded from the analysis due to missing data.