Patient-derived cancer organoid tracking with widefield one-photon redox imaging to assess treatment response

Abstract

MotivationAccessible tools are needed for rapid, non-destructive imaging of patient-derived cancer organoid (PCO) treatment response to accelerate drug discovery and streamline treatment planning for individual patients. AimSegment and track individual PCOs with widefield one-photon redox imaging to extract morphological and metabolic variables of treatment response. ApproachRedox imaging of the endogenous fluorophores, NAD(P)H and FAD, was used to monitor the metabolic state and morphology of PCOs. Redox imaging was performed on a widefield one-photon epifluorescence microscope to evaluate drug response in two colorectal PCO lines. An automated image analysis framework was developed to track PCOs across multiple time points over 48 hours. Variables quantified for each PCO captured metabolic and morphological response to drug treatment, including the optical redox ratio and organoid area. ResultsThe optical redox ratio (NAD(P)H/(FAD+NAD(P)H)) was independent of PCO morphology pre-tieatment. Drugs that induced cell death decreased the optical redox ratio and growth rate compared to control. Multivariate analysis of redox and morphology variables identified distinct PCO sub-populations. Single-organoid tracking improved sensitivity to drug treatment compared to pooled organoid analysis. ConclusionWidefield one-photon redox imaging can monitor metabolic and morphological changes on a single organoid-level, providing an accessible, non-destructive tool to screen drugs in patient-matched samples.