Metabolic Heterogeneity in Patient Tumor-Derived Organoids by Primary Site and Drug Treatment

Abstract

New tools are needed to match cancer patients with effective treatments. Patient-derived organoids offer a high-throughput platform to personalize treatments and discover novel therapies. Currently, methods to evaluate drug response in organoids are limited because they overlook cellular heterogeneity. In this study, non-invasive optical metabolic imaging (OMI) of cellular heterogeneity was characterized in breast cancer (BC) and pancreatic cancer (PC) patient-derived organoids. Baseline heterogeneity was analyzed for each patient, demonstrating that single-cell techniques, such as OMI, are required to capture the complete picture of heterogeneity present in a sample. Treatment-induced changes in heterogeneity were also analyzed, further demonstrating that these measurements greatly complement current techniques that only gauge average cellular response. Finally, OMI of cellular heterogeneity in organoids was evaluated as a predictor of clinical treatment response for the first time. Organoids were treated with the same drugs as the patient's prescribed regimen, and OMI measurements of heterogeneity were compared to patient outcome. OMI distinguished subpopulations of cells with divergent and dynamic responses to treatment in living organoids without the use of labels or dyes. OMI of organoids agreed with long-term therapeutic response in patients. With these capabilities, OMI could serve as a sensitive high-throughput tool to identify optimal therapies for individual patients, and to develop new effective therapies that address cellular heterogeneity in cancer.

Promising Applications of Tumor Spheroids and Organoids for Personalized Medicine

“…Two analyses were used to assess the effect of drugs, which included measuring the cell death ratio and calculating the area under the curve [ 118 ]. An interesting study by Sharick et al used optical metabolic imaging to assess the metabolic response of breast and pancreatic tumor patient-derived organoids in drug screening [ 119 ].…”

The Metabolic Heterogeneity and Flexibility of Cancer Stem Cells

“…Therefore, to more accurately elucidate the metabolism of CSCs, it may be necessary to perform quick analyses in fresh cells immediately after they are taken from the body, or to culture CSCs under conditions close to the original environment. Although it is difficult to reproduce the original tumor microenvironment in vitro, organoid culture technology may enable the development of an effective method to reproduce the heterogeneous structure of tumors [ 143 , 144 , 145 ].…”

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

“…The copyright holder for this preprint this version posted December 21, 2020. ; https://doi.org/10.1101/2020.12.19.423617 doi: bioRxiv preprint caspase 3). [26][27][28][29][30][31][32] However, most studies of autofluorescence in PCOs have used expensive multiphoton and/or fluorescence lifetime imaging microscopes that are not widely available. [26][27][28][29][30][31][32] A recent study by our group used selective plane illumination (light sheet) microscopy (SPIM) to image the autofluorescence of multiple PCOs, but required non-standard culture dishes to accommodate the imaging geometry of the microscope.…”

“…15 In this widefield configuration, redox imaging can capture the optical redox ratio of each pixel along with the morphology of each PCO, but this configuration does not resolve individual cells as in previous two-photon redox imaging studies. [26][27][28][29][30][31][32] However, widefield onephoton redox imaging can rapidly image large populations of PCOs, which is important for characterizing sub-populations of PCOs with variable treatment sensitivities. 9,34 New image analysis methods are needed for widefield one-photon redox imaging that can resolve drug response in heterogenous PCO populations.…”