Evidence of a role for radiation-induced lysosomal damage in non-targeted effects: an experimental and theoretical analysis

Abstract

A well-known DNA-damaging agent and carcinogen, ionizing radiation (IR) can also exert detrimental effects in cells not directly exposed to it, through \"non-targeted effects\" (NTE). Whilst NTE are known to contribute to radiation-induced damage, their mechanism of induction and propagation remains incompletely understood. To investigate the possible role of lysosomes, key subcellular organelles, in NTE we used acridine orange uptake and relocation methods to monitor lysosomal permeability in irradiated and bystander human fibroblasts. As a potential mediator of lysosomal changes, oxidative stress was measured using the H2DCFDA assay for total reactive oxygen species (ROS). IR was found to induce significant lysosomal permeability in the first hour post irradiation, with reduced permeability persisting up to 24 hours. This occurred in conjunction with an increase in ROS in directly irradiated cells, in contrast with a decrease in ROS in bystander cells. Based on these observations we constructed a simple mathematical model of ROS-induced lysosomal damage, based on a bistable mechanism where a sufficiently strong IR insult can shift a cell from a low ROS, high lysosome to a high ROS, low lysosome state. This has profound cellular implications in radiation response and advances our understanding for the sub-cellular involvement in non-targeted effects.