Title:
Advancing Drug Discovery through 3D Cell Models: Unveiling Novel Hits and Mechanisms
Autor :
Nicolas Ancellin*, Olivier Duchamp, Kenny Herry, Nicolas Legrand, Christophe Parsy
Oncodesign Services – 20 rue Jean Mazen 21000 Dijon
Abstract :
Traditional 2D cell cultures, while a standard in research, struggle to replicate in vivo complexity. The integration of 3D cell models not only offers ethical and practical advantages but also expedites drug development by reducing the need for animal experimentation. With enhanced experimental material and the ability for genetic manipulation, these models enable exploration of molecular mechanisms, leading to novel therapeutic insights
In this study, we present an innovative approach to hit identification and confirmation using spheroids and patient-derived xenograft organoids (PDXOs). Spheroids, in contrast to 2D cultures, more faithfully mimic cell-cell interactions and gradients of nutrients and oxygen found in tumors. Leveraging a focused chemical library, a phenotypic screen was conducted on two triple-negative breast cancer cell lines. The results not only revealed new hit identification but also unraveled a novel potential kinase target, showcasing the power of 3D cell models in drug screening.
Advancing our research, we employed patient-derived xenograft organoids (PDXOs), a convergence of PDX models and organoid technology. PDXOs retain the genetic diversity and complexity of tumors, providing a translational platform for medium-throughput drug testing. The multifaceted applications of PDXOs in phenotypic screens emerged as a transformative approach, not only aiding in identifying potential drug leads but also serving as a critical step in refining chemistry for lead optimization and unraveling new targets.
To validate the utility of PDXOs, we generated pancreatic ductal adenocarcinoma (PDAC) PDXOs from four patients and subjected them to a proliferation assay using gold standard treatment. Correlation with in vivo treatment using Irinotecan was established, validating the predictive power of the 3D model. Additionally, a small exclusive Benzoxaboroles compound library was screened. Benzoxaboroles, a class of organoboron compounds, exhibit a unique structure that enables them to engage in various interactions with biological molecules, such as coordination to Oxygen and Nitrogen and engaging in reversible covalent and non-covalent binding interactions with protein amino acids, particularly serine. Results discover hits that responded differently based on the KRAS mutational status of the PDXOs.
An additional advantage of a 3D cell system, which closely mimics the patient, is the ability to perform target validation — experimental investigations confirming and assessing the biological relevance and potential therapeutic impact of a specific molecular target for drug development. We exemplified this process by implementing CRISPR-Cas9 knockdown technology in PDAC PDXO, targeting a specific kinase to gain insights into its role in proliferation.
In conclusion, our findings highlight the importance of 3D cell models in drug discovery. These approaches enhance understanding of individual patient responses, facilitate in vivo studies, and bridge the gap between laboratory discoveries and clinical applications. Spheroids and PDX-Organoids offer a realistic microenvironment, enabling accurate phenotypic screens, reliable target validation, and a foundation for hit identification and confirmation.
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