High-Throughput Imaging Assay for Drug Screening of 3D Prostate Cancer Organoids

Abstract

New treatments are required for advanced prostate cancer; however, there are fewer preclinical models of prostate cancer than other common tumor types to test candidate therapeutics. One opportunity to increase the scope of preclinical studies is to grow tissue from patient-derived xenografts (PDXs) as organoid cultures. Here we report a scalable pipeline for automated seeding, treatment and an analysis of the drug responses of prostate cancer organoids. We established organoid cultures from 5 PDXs with diverse phenotypes of prostate cancer, including castrate-sensitive and castrate-resistant disease, as well as adenocarcinoma and neuroendocrine pathology. We robotically embedded organoids in Matrigel in 384-well plates and monitored growth via brightfield microscopy before treatment with poly ADP-ribose polymerase inhibitors or a compound library. Independent readouts including metabolic activity and live-cell imaging–based features provided robust measures of organoid growth and complementary ways of assessing drug efficacy. Single organoid analyses enabled in-depth assessment of morphological differences between patients and within organoid populations and revealed that larger organoids had more striking changes in morphology and composition after drug treatment. By increasing the scale and scope of organoid experiments, this automated assay complements other patient-derived models and will expedite preclinical testing of new treatments for prostate cancer.

Combined Systemic Drug Treatment with Proton Therapy: Investigations on Patient-Derived Organoids

“…The translation to organoids is challenging, especially for large-scale experiments, as they grow in a matrix and display heterogeneous morphology. To this end, image-based analyses using advanced methods including machine-learning based approaches may have the potential to solve these issues [ 54 ].…”

Label-Free Imaging Analysis of Patient-Derived Cholangiocarcinoma Organoids after Sorafenib Treatment

“…Our results show that this pipeline can be easily adapted to iCCAOs. This label-free imaging approach offers great potential for high-throughput screening of patient-derived organoid cultures [ 22 , 23 , 40 , 41 ]. However, detailed data from other disciplines such as proteomics [ 42 ] and transcriptomics [ 43 ] are needed to ascribe the morphologic changes observed over time to specific underlying cellular processes.…”

Mimicking the Natural Basement Membrane for Advanced Tissue Engineering

“…Geltrex has been used to maintain and scale up human pluripotent stem cells (hPSC) that can be used for downstream differentiation for therapeutic applications . These naturally derived matrices have also been used to support and maintain various organoids including intestine, brain, inner ear, prostate, and lung. − However, although these matrices are of natural origin, there are limitations to their application as BM mimics such as precise working temperatures (4 °C) to prevent gelling, tuning their biochemical or biomechanical properties without influencing other material attributes, , and a lack of knowledge of their molecular composition. These factors tend to limit their translation. , However, the group of Mikhail et al has used Matrigel together with collagen I to produce 3D hydrogels that support mini-gut culture.…”