Analysis of lncRNA-miRNA-mRNA expression pattern in heart tissue after total body radiation in a mouse model

Abstract

Background Radiation therapy is integral to effective thoracic cancer treatments, but its application is limited by sensitivity of critical organs such as the heart. The impacts of acute radiation-induced damage and its chronic effects on normal heart cells are highly relevant in radiotherapy with increasing lifespans of patients. Biomarkers for normal tissue damage after radiation exposure, whether accidental or therapeutic, are being studied as indicators of both acute and delayed effects. Recent research has highlighted the potential importance of RNAs, including messenger RNAs (mRNAs), microRNAs (miRNAs), and long non-coding RNAs (lncRNAs) as biomarkers to assess radiation damage. Understanding changes in mRNA and non-coding RNA expression will elucidate biological pathway changes after radiation. Methods To identify significant expression changes in mRNAs, lncRNAs, and miRNAs, we performed whole transcriptome microarray analysis of mouse heart tissue at 48 h after whole-body irradiation with 1, 2, 4, 8, and 12 Gray (Gy). We also validated changes in specific lncRNAs through RT-qPCR. Ingenuity Pathway Analysis (IPA) was used to identify pathways associated with gene expression changes. Results We observed sustained increases in lncRNAs and mRNAs, across all doses of radiation. Alas2, Aplnr, and Cxc3r1 were the most significantly downregulated mRNAs across all doses. Among the significantly upregulated mRNAs were cell-cycle arrest biomarkers Gdf15, Cdkn1a, and Ckap2. Additionally, IPA identified significant changes in gene expression relevant to senescence, apoptosis, hemoglobin synthesis, inflammation, and metabolism. LncRNAs Abhd11os, Pvt1, Trp53cor1, and Dino showed increased expression with increasing doses of radiation. We did not observe any miRNAs with sustained up- or downregulation across all doses, but miR-149-3p, miR-6538, miR-8101, miR-7118-5p, miR-211-3p, and miR-3960 were significantly upregulated after 12 Gy. Conclusions Radiation-induced RNA expression changes may be predictive of normal tissue toxicities and may indicate targetable pathways for radiation countermeasure development and improved radiotherapy treatment plans.

Microarray analysis identifies coding and non-coding RNA markers of liver injury in whole body irradiated mice

“…All three have previously been highlighted as predictive markers of radiation injury in murine models 38 , 58 – 61 . Cdkn1a has previously been reported as a marker of irradiation in cancer patients undergoing WBI and in murine models from our lab and others 25 , 35 , 62 .…”

“…Research from our lab and others have highlighted the radiation response of multiple lncRNA. Some of these lncRNA are otherwise uncharacterized, while others, such as Trp53cor1 and Dino , are relatively well known 25 – 28 . MicroRNA (miRNA), non-coding RNA of roughly 22 nucleotides, exert their functions by hybridizing to complementary sequences in the 3’ UTRs of mRNAs, which lead to either RNA degradation or translation inhibition 29 .…”

Non-coding RNAs in cancer therapy-induced cardiotoxicity: Mechanisms, biomarkers, and treatments

“…A recent study screened mouse cardiac tissue for the changes in mRNAs, miRNAs, and lncRNAs levels that predict radiation-induced heart damage after exposing mice to 1, 2, 4, 8, and 12 Gray (Gy) whole-body irradiation for 48 h ( 177 ). The study found that the expression alterations in numerous mRNAs and ncRNAs at different doses, such as LncRNA Abhd11os, Pvt1, Trp53cor1, and Dino, rose as the radiation dose increased.…”

Biomarkers to Predict Lethal Radiation Injury to the Rat Lung

“…However, despite decades of research, a gap in this field remains. Biological molecules that have been pursued include DNA [ 2 , 3 ], multiple species of RNA such as long non-coding RNA (lncRNA) [ 4 ], miRNA [ 5 , 6 ], mRNA [ 4 , 7 ], proteins and protein arrays [ 8 , 9 , 10 ], advanced multi-omic platforms [ 11 ] and others [ 12 ]. Essential to bridging this gap is establishing representative animal models and conducting relevant clinical trials.…”