High-Throughput Imaging Assay for Drug Screening of 3D Prostate Cancer Organoids
Nicholas Choo1, Susanne Ramm2,3, Jennii Luu3
1Monash Partners Comprehensive Cancer Consortium, Monash Biomedicine Discovery Institute Cancer Program, Prostate Cancer Research Group, Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC, Australia.
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.
Insights
Researchers developed an automated pipeline to grow and test prostate cancer organoids from patient tumors. This scalable method accelerates the preclinical testing of new drug treatments for advanced prostate cancer.
Area of Science:
- Oncology
- Biotechnology
- Drug Discovery
Background:
- Advanced prostate cancer necessitates novel therapeutic strategies.
- Limited preclinical models hinder the development of new prostate cancer treatments.
- Patient-derived xenografts (PDXs) offer a valuable source for preclinical research.
Purpose of the Study:
- To establish a scalable, automated pipeline for prostate cancer organoid culture and drug response analysis.
- To assess the efficacy of novel therapeutic agents using diverse prostate cancer organoid models.
- To enhance the scope and efficiency of preclinical drug testing for prostate cancer.
Main Methods:
- Established organoid cultures from five patient-derived xenografts (PDXs) representing diverse prostate cancer phenotypes.
- Utilized robotic systems for automated organoid seeding, Matrigel embedding, and treatment in 384-well plates.
- Employed brightfield microscopy, metabolic activity assays, and live-cell imaging for robust drug response assessment.
- Conducted single organoid analyses to evaluate morphological changes and drug-induced effects.
Main Results:
- Successfully cultured prostate cancer organoids from PDXs with varied disease characteristics (castrate-sensitive, castrate-resistant, adenocarcinoma, neuroendocrine).
- Demonstrated robust drug response measurements using independent readouts like metabolic activity and imaging features.
- Identified distinct morphological changes in larger organoids post-treatment, providing insights into drug efficacy.
- Validated the automated pipeline's capacity for scalable and comprehensive organoid-based drug screening.
Conclusions:
- The developed automated pipeline provides a scalable and efficient platform for prostate cancer organoid drug screening.
- This approach significantly enhances the utility of patient-derived organoids in preclinical research.
- The automated assay will expedite the identification and validation of new treatments for advanced prostate cancer.
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