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Utilizing Functional Genomics Screening to Identify Potentially Novel Drug Targets in Cancer Cell Spheroid Cultures
Published on: December 26, 2016
Utilizing Functional Genomics Screening to Identify Potentially Novel Drug Targets in Cancer Cell Spheroid Cultures
Eamonn Morrison1, Patty Wai1, Andri Leonidou1
1The Breast Cancer Now Toby Robins Research Centre, Division of Breast Cancer, The Institute of Cancer Research; Division of Molecular Pathology, The Institute of Cancer Research.
Abstract:
The identification of functional driver events in cancer is central to furthering our understanding of cancer biology and indispensable for the discovery of the next generation of novel drug targets. It is becoming apparent that more complex models of cancer are required to fully appreciate the contributing factors that drive tumorigenesis in vivo and increase the efficacy of novel therapies that make the transition from pre-clinical models to clinical trials. Here we present a methodology for generating uniform and reproducible tumor spheroids that can be subjected to siRNA functional screening. These spheroids display many characteristics that are found in solid tumors that are not present in traditional two-dimension culture. We show that several commonly used breast cancer cell lines are amenable to this protocol. Furthermore, we provide proof-of-principle data utilizing the breast cancer cell line BT474, confirming their dependency on amplification of the epidermal growth factor receptor HER2 and mutation of phosphatidylinositol-4,5-biphosphate 3-kinase (PIK3CA) when grown as tumor spheroids. Finally, we are able to further investigate and confirm the spatial impact of these dependencies using immunohistochemistry.
Insights
This study introduces a reproducible method for creating tumor spheroids for functional screening. These 3D models better mimic solid tumors, confirming key cancer gene dependencies like HER2 and PIK3CA in breast cancer cells.
Area of Science:
- Oncology
- Cancer Biology
- Drug Discovery
Background:
- Identifying functional driver events is crucial for understanding cancer biology and developing new drug targets.
- In vivo tumorigenesis and therapeutic efficacy require more complex cancer models than traditional 2D cultures.
- Advanced models are needed to bridge pre-clinical findings to clinical trials.
Purpose of the Study:
- To develop a reproducible methodology for generating uniform tumor spheroids.
- To enable functional screening using siRNA in these 3D tumor models.
- To validate the utility of tumor spheroids in studying cancer driver events.
Main Methods:
- Generation of uniform and reproducible tumor spheroids.
- siRNA functional screening assays performed on tumor spheroids.
- Immunohistochemistry to analyze spatial impact of genetic dependencies.
Main Results:
- The developed protocol is applicable to common breast cancer cell lines.
- Tumor spheroids exhibit characteristics of solid tumors, unlike 2D cultures.
- BT474 breast cancer cells showed dependency on epidermal growth factor receptor (HER2) amplification and phosphatidylinositol-4,5-biphosphate 3-kinase (PIK3CA) mutation in spheroids.
Conclusions:
- Tumor spheroids provide a more relevant model for studying cancer driver events.
- This methodology facilitates functional screening and validation of drug targets.
- The study confirms the spatial impact of HER2 and PIK3CA in breast cancer models.
