Data-Driven Identification of Biomarkers for In Situ Monitoring of Drug Treatment in Bladder Cancer Organoids

Abstract

Three-dimensional (3D) organoid culture recapitulating patient-specific histopathological and molecular diversity offers great promise for precision medicine in cancer. In this study, we established label-free imaging procedures, including Raman microspectroscopy (RMS) and fluorescence lifetime imaging microscopy (FLIM), for in situ cellular analysis and metabolic monitoring of drug treatment efficacy. Primary tumor and urine specimens were utilized to generate bladder cancer organoids, which were further treated with various concentrations of pharmaceutical agents relevant for the treatment of bladder cancer (i.e., cisplatin, venetoclax). Direct cellular response upon drug treatment was monitored by RMS. Raman spectra of treated and untreated bladder cancer organoids were compared using multivariate data analysis to monitor the impact of drugs on subcellular structures such as nuclei and mitochondria based on shifts and intensity changes of specific molecular vibrations. The effects of different drugs on cell metabolism were assessed by the local autofluorophore environment of NADH and FAD, determined by multiexponential fitting of lifetime decays. Data-driven neural network and data validation analyses (k-means clustering) were performed to retrieve additional and non-biased biomarkers for the classification of drug-specific responsiveness. Together, FLIM and RMS allowed for non-invasive and molecular-sensitive monitoring of tumor-drug interactions, providing the potential to determine and optimize patient-specific treatment efficacy.

Efficacy and Safety of Resveratrol in the Treatment of Bladder Cancer Derived Organoids

“…When spheroids reached appropriate sizes (80-120 μm), they were treated by RES (0, 10 × 10 −6 , 25 × 10 −6 , 50 × 10 −6 , 75 × 10 −6 , 100 × 10 −6 , 150 × 10 −6 , and 200 × 10 −6 m) and EPI (0, 0.01 × 10 −6 , 0.1 × 10 −6 , 0.5 × 10 −6 , 1 × 10 −6 , 2 × 10 −6 , 5 × 10 −6 , 10 × 10 −6 , and 50 × 10 −6 m) respectively for 96 h. Each of the groups was set in triplicate. [27,45] After 96 h of treatment, 10 μL of Cell Counting Kit-3D (CCK-3D, Beyotime, Shanghai, China) was added to each well and incubated at 37 °C for 30 min to determine the absorbance at 450 nm. Half-inhibitory concentration (IC 50 ) was calculated to determine the antitumor efficacy of the tested drug.…”

Preclinical models for bladder cancer therapy research

“…Efforts were also made to develop novel methods for the accurate analysis of 3-dimensional BC models. Becker et al established a label-free and noninvasive imaging procedure to monitor drug response combining Raman microspectroscopy (RMS) and fluorescence lifetime imaging microscopy (FLIM) [ 21 ]. Zhang et al developed a deep learning model for the evaluation of the growth status of organoids called AU2Net (Attention and Cross U2Net) [ 22 ].…”

Role of the tumour microenvironment in bladder cancer pathogenesis and value of the reverse translational approach: a tale of two species

“…These can be generated from surgical specimens, biopsies and ‘patients’ body fluids, such as samples obtained from fine‐needle aspiration 180–182 . Notably, in humans and dogs with MIBC, PDOs have been successfully generated from urine samples in a non‐invasive manner 183,184 . Moreover, 3D bladder cancer organoids derived from the primary tumour and urine specimens were used for testing various pharmaceutical candidates, for example, cisplatin and venetoclax, using different concentrations and measuring direct cellular responses as outputs 183 …”

“…[180][181][182] Notably, in humans and dogs with MIBC, PDOs have been successfully generated from urine samples in a non-invasive manner. 183,184 Moreover, 3D bladder cancer organoids derived from the primary tumour and urine specimens were used for testing various pharmaceutical candidates, for example, cisplatin and venetoclax, using different concentrations and measuring direct cellular responses as outputs. 183 These PDOs can adequately model the cellular complexity of the native tumours, while more conventional 2D cell lines typically fail to recapitulate.…”

“…183,184 Moreover, 3D bladder cancer organoids derived from the primary tumour and urine specimens were used for testing various pharmaceutical candidates, for example, cisplatin and venetoclax, using different concentrations and measuring direct cellular responses as outputs. 183 These PDOs can adequately model the cellular complexity of the native tumours, while more conventional 2D cell lines typically fail to recapitulate. 185,186 In addition to mimicking most tumour characteristics, including metabolic features and immunosuppressive capabilities, 186 PDOs can capture the interactions between cellular elements and the tumour matrix, which influence phenotypic features and signalling pathways.…”