Method Profile
Track individual organoids during and after drug exposure, alongside bulk ATP and endpoint size measurements.
Compare complementary pharmacology readouts and explore associations with chemotherapy or chemoradiation response.
Patient-derived primary and metastatic colorectal cancer organoids. The chemotherapy and rectal cancer collections include repeat samples from some patients.
48-hour drug exposure followed by 48 hours without drug. Individual-organoid diameter change and optical redox measurements from NAD(P)H and FAD are compared with bulk and endpoint readouts.
Treatment conditions
5-fluorouracil 10 μM, oxaliplatin 5 μM, SN38 1.5 nM and radiation 2 Gy in specified combination arms.
Readouts
Individual-organoid diameter change; widefield optical redox measurements from NAD(P)H and FAD; whole-well CellTiter-Glo and endpoint diameter comparators.
Analyze change within the same organoid rather than pooling the well. Compare growth and metabolic signals, then examine sensitive, intermediate and resistant classifications against clinical response.
Readout comparison and exploratory clinical concordance.
Initial FOLFOX collection: 13 samples from 13 patients. Rectal collection: 17 samples from 13 patients. Clinical analyses covered 20 chemotherapy subjects and 11 treatment-naive chemoradiation subjects.
Untreated organoids; endpoint diameter; ATP assay; optical redox; patient radiological/pathological response.
Complementary readouts detected different responses. Combined diameter/redox sensitive classification included 12 subjects: 92% had partial or complete response and one had stable disease. Half of resistant classifications still had partial responses.
Longitudinal diameter and metabolic imaging detected complementary responses in colorectal cancer organoids. Sensitive classifications often aligned with patient response, while resistant classifications still included clinical responders.
