Accelerating precision anti-cancer therapy by time-lapse and label-free 3D tumor slice culture platform

Abstract

The feasibility of personalized medicine for cancer treatment is largely hampered by costly, labor-intensive and time-consuming models for drug discovery. Herein, establishing new pre-clinical models to tackle these issues for personalized medicine is urgently demanded. Methods: We established a three-dimensional tumor slice culture (3D-TSC) platform incorporating label-free techniques for time-course experiments to predict anti-cancer drug efficacy and validated the 3D-TSC model by multiphoton fluorescence microscopy, RNA sequence analysis, histochemical and histological analysis. Results : Using time-lapse imaging of the apoptotic reporter sensor C3 (C3), we performed cell-based high-throughput drug screening and shortlisted high-efficacy drugs to screen murine and human 3D-TSCs, which validate effective candidates within 7 days of surgery. Histological and RNA sequence analyses demonstrated that 3D-TSCs accurately preserved immune components of the original tumor, which enables the successful achievement of immune checkpoint blockade assays with antibodies against PD-1 and/or PD-L1. Label-free multiphoton fluorescence imaging revealed that 3D-TSCs exhibit lipofuscin autofluorescence features in the time-course monitoring of drug response and efficacy. Conclusion : This technology accelerates precision anti-cancer therapy by providing a cheap, fast, and easy platform for anti-cancer drug discovery.

An In Vivo Fluorescence Resonance Energy Transfer-Based Imaging Platform for Targeted Drug Discovery and Cancer Therapy

“…In a previous study, HeLa cells employing Förster resonance energy transfer (FRET) technique was used to detect caspase-3 activation, which affected energy transfer between the cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) and lowered the FRET ratio of the probe ( Luo et al, 2001 ). In our earlier study, we also used FRET to visualize the dynamics of doxorubicin (DOX) treated single cells and test the pharmacodynamics of drug treated tumor slices in a 3D-tumor slice culture (3D-TSCs) model ( Xing et al, 2021 ). However, whether the FRET technique can be applied to animal models and drug evaluation remains yet to be explored.…”

Tumor heterogeneity reshapes the tumor microenvironment to influence drug resistance

“…Some novel and effective preclinical models could be implemented to assess drug responses and efficacy. For example, a three-dimensional tumor slice cultures (3D-TSC) platform ( Figure 4 ) was reported recently, which combines histochemical staining, transgenic fluorescent reporter, and label-free metabolic imaging approaches in a time course to evaluate drug efficacy to accelerate precision drug screening for cancer therapy 99 . As 3D-TSC not only maintains important original tumor features, such as tumor heterogeneity, gene expression, and cell architecture but also maintain all immune components, it should be a powerful model for testing the drug sensitivity.…”

“…On the one hand, personalized medicine should be applied here to continually evaluate the dynamic changes in tumor heterogeneity levels and TME complexity levels to better choose more appropriate drugs for patients. On the other hand, some novel and effective preclinical models, such as the 3D-TSC platform, could be implemented to assess drug responses and efficacy 99 . 3D-TSC is a good model for observing dynamic changes in the cell population during drug treatment.…”

Recent advances in organotypic tissue slice cultures for anticancer drug development

“…It maximally retains many aspects of the original tumors, including inter-tumor and intratumor heterogeneity, TME, morphology, cellular-stromal interplay, and complexity [24,[41][42][43][44][45]. The 3D-TSC platform experiences expanding advantages in these regards [27]. First, the slice kept growing for at least 10 days with a gradually increasing number of total live cells (Figure 3A-B).…”

“…The organotypic tissue slice culture platform is also pragmatic for anticancer drug discovery owing to the integration of the alike in vivo microanatomy and the easy manipulation of in vitro work [24][25][26]. We recently developed a 3D tumor slice culture (3D-TSC) platform that incorporates a lipofuscin autofluorescence feature for the time-course monitoring of drug responses [27]. Compared with other mainstreams, the 3D-TSC platform presents a comparable potential for anticancer drug development (Table 1).…”