ArticlebioRxiv : the preprint server for biology2024
Wide-field optical redox imaging with leading-edge detection for assessment of patient-derived cancer organoids.
Article in bioRxiv : the preprint server for biology, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Patient-derived cancer organoids (PDCOs) are a valuable model to recapitulate human disease in culture with important implications for drug development. However, current methods for assessing PDCOs are limited. Label-free imaging methods are a promising tool to measure organoid level heterogeneity and rapidly screen drug response in PDCOs. The aim of this study was to assess and predict PDCO response to treatments based on mutational profiles using label-free wide-field optical redox imaging (WF ORI). WF ORI provides organoid-level measurements of treatment response without labels or additional reagents by measuring the autofluorescence intensity of the metabolic co-enzymes NAD(P)H and FAD. The optical redox ratio is defined as the fluorescence intensity of [NAD(P)H / NAD(P)H +FAD] which measures the oxidation-reduction state of PDCOs. We have implemented WF ORI and developed novel leading-edge analysis tools to maximize the sensitivity and reproducibility of treatment response measurements in colorectal PDCOs. Leading-edge analysis improves sensitivity to redox changes in treated PDCOs (GΔ = 1.462 vs GΔ = 1.233). Additionally, WF ORI resolves FOLFOX treatment effects across all PDCOs better than two-photon ORI, with ∼7X increase in effect size (GΔ = 1.462 vs GΔ = 0.189). WF ORI distinguishes metabolic differences based on driver mutations in CRC PDCOs identifying Statement of Significance: Label-free wide-field optical redox imaging of patient-derived cancer organoids enables rapid, sensitive, and reproducible measurements of treatment response and heterogeneity that will impact patient management, clinical trials, and preclinical drug development.
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