Modeling Prostate Cancer Treatment Responses in the Organoid Era: 3D Environment Impacts Drug Testing

Abstract

Organoid-based studies have revolutionized in vitro preclinical research and hold great promise for the cancer research field, including prostate cancer (PCa). However, experimental variability in organoid drug testing complicates reproducibility. For example, we observed PCa organoids to be less affected by cabazitaxel, abiraterone and enzalutamide as compared to corresponding single cells prior to organoid assembly. We hypothesized that three-dimensional (3D) organoid organization and the use of various 3D scaffolds impact treatment efficacy. Live-cell imaging of androgen-induced androgen receptor (AR) nuclear translocation and taxane-induced tubulin stabilization was used to investigate the impact of 3D scaffolds, spatial organoid distribution and organoid size on treatment effect. Scaffolds delayed AR translocation and tubulin stabilization, with Matrigel causing a more pronounced delay than synthetic hydrogel as well as incomplete tubulin stabilization. Drug effect was further attenuated the more centrally organoids were located in the scaffold dome. Moreover, cells in the organoid core revealed a delayed treatment effect compared to cells in the organoid periphery, underscoring the impact of organoid size. These findings indicate that analysis of organoid drug responses needs careful interpretation and requires dedicated read-outs with consideration of underlying technical aspects.

Let’s Go 3D! New Generation of Models for Evaluating Drug Response and Resistance in Prostate Cancer

“…This model is also known as the liquid-overlay method, as the cells are first embedded in the matrix, and then the pellet is overlaid with the culture media. However, some disadvantages are the undefined composition of natural gels and the structural modification over time [ 69 , 70 , 71 , 72 , 73 , 74 ], as well as their impact on the lower penetration of the drugs to the destination cells, leading to lower drug efficacy [ 75 ].…”

Mimicking the Natural Basement Membrane for Advanced Tissue Engineering

“…These factors tend to limit their translation. 180 , 181 However, the group of Mikhail et al has used Matrigel together with collagen I to produce 3D hydrogels that support mini-gut culture. Collagen I provided mechanical support, and Matrigel offered key components of cell-adhesion present in BM, and when they were used in a perfusable microdevice, they offered support to intestinal stem cells for organoid formation.…”

“…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. Collagen I provided mechanical support, and Matrigel offered key components of cell-adhesion present in BM, and when they were used in a perfusable microdevice, they offered support to intestinal stem cells for organoid formation …”

How to build a tumor: An industry perspective

“…Live cell imaging, transcriptomic and proteomic analyses will be required to determine the most efficacious candidate. 60 .…”