Abstract 3313: Single-cell level absorbed dose assessment and radiosensitivity modeling for alpha-emitter radiopharmaceutical therapy
Authors: Bastiaannet Remco , Liatsou Ioanna , Hobbs Robert , Sgouros George 
Published: June 2022

Abstract
The radiobiological response to alpha-emitter radiopharmaceutical therapy (αRPT) is often studied in cell monolayers. In these experiments, the goal is to establish the relationship between absorbed dose and cell survival probability. However, absorbed dose is not readily known and needs to be estimated. Current models to estimate this commonly idealize cells as biologically inert spheres (geometric model). This results in inaccurate and non-generalizable results, which could hamper the rigorous study of the underlying radiobiology. The purpose of this study was to estimate the variability in absorbed dose on a single-cell level by combining 3D measurements of cell geometries, as well as the dynamics of carrier molecule binding and trafficking in individual cells with full physics simulations of the alpha emissions. This comprehensive integration of the biological and physical aspects allows for a more accurate way to model cell survival in these αRPT experiments. Live cells were imaged on a confocal microscope. Experimental conditions of previous cell survival experiments with 212Pb on NT2.5 HER2+ breast cancer cells were replicated. A relevant antibody was tagged with AF488. 3D time lapses of membrane binding kinetics and internalization were recorded. All cells were segmented into nuclei, membrane and cytosol compartments using a purpose-build algorithm. Pharmacokinetic models were fit to the temporal antibody signals, which enabled validation with experimental binding assays. Monte Carlo simulations using the exact antibody locations in each time frame allowed for the precise estimation of dose rate over time. Single-cell absorbed dose estimates were used to model previously obtained cell survival curves. Over 300 cells were measured between 0 and 26 hours post incubation. A large range in absorbed doses was observed (coefficient of variation 0.74). The median contribution of membrane-bound activity to absorbed dose was in agreement with geometric models (error less than 6%). However the contribution of antibody internalization and perinuclear trafficking to absorbed dose varied widely between cells and over time. Cell clustering contributed 46% of the total dose, and was 6x higher than in the geometric model. Applying this to previous cell survival data yielded an estimated radiosensitivity kappa of 7.1 (geometric model: 2.8). Cell clustering has a larger, and cell geometry has a smaller impact than is assumed in current models. Perinuclear trafficking of internalized Ab positively impacts cell nucleus absorbed dose, which is typically ignored. Dose variability should be included in radiosensitivity modeling. More accurate absorbed dose estimations which result from a better understanding of the different contributing factors, will improve generalizability of the radiobiological models established in these cell monolayer experiments to more complex models.
Citation Format: Remco Bastiaannet, Ioanna Liatsou, Robert Hobbs, George Sgouros. Single-cell level absorbed dose assessment and radiosensitivity modeling for alpha-emitter radiopharmaceutical therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3313.