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High-throughput dynamic BH3 profiling may quickly and accurately predict effective therapies in solid tumors
Patrick D Bhola1,2, Eman Ahmed1, Jennifer L Guerriero1
1Dana-Farber Cancer Institute, Boston, MA 02215, USA.
Abstract:
Despite decades of effort, the sensitivity of patient tumors to individual drugs is often not predictable on the basis of molecular markers alone. Therefore, unbiased, high-throughput approaches to match patient tumors to effective drugs, without requiring a priori molecular hypotheses, are critically needed. Here, we improved upon a method that we previously reported and developed called high-throughput dynamic BH3 profiling (HT-DBP). HT-DBP is a microscopy-based, single-cell resolution assay that enables chemical screens of hundreds to thousands of candidate drugs on freshly isolated tumor cells. The method identifies chemical inducers of mitochondrial apoptotic signaling, a mechanism of cell death. HT-DBP requires only 24 hours of ex vivo culture, which enables a more immediate study of fresh primary tumor cells and minimizes adaptive changes that occur with prolonged ex vivo culture. Effective compounds identified by HT-DBP induced tumor regression in genetically engineered and patient-derived xenograft (PDX) models of breast cancer. We additionally found that chemical vulnerabilities changed as cancer cells expanded ex vivo. Furthermore, using PDX models of colon cancer and resected tumors from colon cancer patients, our data demonstrated that HT-DBP could be used to generate personalized pharmacotypes. Thus, HT-DBP appears to be an ex vivo functional method with sufficient scale to simultaneously function as a companion diagnostic, therapeutic personalization, and discovery tool.
Insights
High-throughput dynamic BH3 profiling (HT-DBP) screens drugs on patient tumors ex vivo. This functional assay predicts effective cancer treatments and personalizes therapy, overcoming limitations of molecular markers alone.
Area of Science:
- Oncology
- Biotechnology
- Pharmacology
Background:
- Predicting patient tumor sensitivity to drugs solely via molecular markers remains challenging.
- There is a critical need for unbiased, high-throughput methods to match tumors with effective drugs without prior molecular assumptions.
Purpose of the Study:
- To present an improved high-throughput dynamic BH3 profiling (HT-DBP) method for drug screening on patient tumors.
- To demonstrate HT-DBP's utility as a companion diagnostic, therapeutic personalization, and drug discovery tool.
Main Methods:
- HT-DBP is a microscopy-based, single-cell assay for screening hundreds to thousands of drugs on fresh tumor cells.
- The assay identifies drugs that induce mitochondrial apoptotic signaling, a cell death pathway.
- It requires only 24 hours of ex vivo culture to minimize adaptive changes in tumor cells.
Main Results:
- HT-DBP identified effective compounds that induced tumor regression in breast cancer models (genetically engineered and patient-derived xenografts).
- Drug sensitivities of cancer cells were observed to change during ex vivo culture.
- Personalized pharmacotypes were generated using HT-DBP in colon cancer models and patient samples.
Conclusions:
- HT-DBP is a scalable, ex vivo functional assay for immediate drug sensitivity testing on fresh tumor cells.
- The method can simultaneously serve as a companion diagnostic, therapeutic personalization tool, and drug discovery platform.
- HT-DBP addresses the need for predictive biomarkers by directly assessing drug response in patient tumors.
