Spatial gene expression changes in the mouse heart following base-targeted irradiation

Abstract

Purpose: Radiation cardiotoxicity (RC) is a clinically significant side effect of treatment for patients with thoracic malignancies. Clinical studies in lung cancer have indicated that heart substructures are not uniformly radiosensitive, and that dose to the heart base drives RC. In this study, we aimed to characterise late changes in gene expression using spatial transcriptomics in a mouse model of base regional radiosensitivity. Materials and methods: An aged female C57BL/6 mouse was irradiated with 16 Gy delivered to the cranial third of the heart using a 6 × 9 mm parallel opposed beam geometry on a small animal radiation research platform, and a second mouse was sham-irradiated. Following echocardiography, whole hearts were collected at 30 weeks for spatial transcriptomic analysis in order to map gene expression changes occurring in different regions of the partially irradiated heart. Cardiac regions were manually annotated on the capture slides and the gene expression profiles compared across different regions. Results: Ejection fraction was reduced at 30 weeks following a 16 Gy irradiation to the heart base, compared with the sham-irradiated control. There were markedly more significant gene expression changes within the irradiated regions compared with non-irradiated regions. Variation was observed in the transcriptomic impact of radiation on different cardiac base structures. eg. between the right atrium (n=86 dysregulated genes), left atrium (n=96 dysregulated genes), and the vasculature (n=129 dysregulated genes). Disrupted biological processes spanned extracellular matrix, circulatory, neuronal and contractility activities. Conclusions: This is the first study to report spatially resolved gene expression changes in irradiated tissues. Examination of the regional radiation response in the heart can help to further our understanding of the cardiac base's radiosensitivity, and support the development of actionable targets for pharmacological intervention, and biologically relevant dose constraints.