Related Experiment Video
Updated: Dec 29, 2025
Profiling Sensitivity to Targeted Therapies in EGFR-Mutant NSCLC Patient-Derived Organoids
Published on: November 22, 2021
Use of Ex Vivo Patient-Derived Tumor Organotypic Spheroids to Identify Combination Therapies for HER2 Mutant
Elena Ivanova1,2, Mari Kuraguchi1,2, Man Xu1,2
1Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, Massachusetts.
Purpose:
Evaluating drug responses using primary patient-derived cells ex vivo represents a potentially rapid and efficient approach to screening for new treatment approaches. Here, we sought to identify neratinib combinations in HER2 mutant non-small cell lung cancer (NSCLC) patient xenograft-derived organotypic spheroids (XDOTS) using a short-term ex vivo system.
Experimental Design:
We generated two HER2-mutant NSCLC PDX models [DFCI359 (HER2 exon19 755_757LREdelinsRP) and DFCI315 (HER2 exon20 V777_G778insGSP)] and used the PDX tumors to generate XDOTS. Tumor spheroids were grown in a microfluidic device and treated ex vivo with neratinib-based drug combinations. Live/dead quantification was performed by dual-labeling deconvolution fluorescence microscopy. The most efficacious ex vivo combination was subsequently validated in vivo using the DFCI359 and DFCI315 PDXs and a HER2 YVMA genetically engineered mouse model.
Results:
Both neratinib and afatinib, but not gefitinib, induced cell death in DFCI359 XDOTS. The combinations of neratinib/trastuzumab and neratinib/temsirolimus enhanced the therapeutic benefit of neratinib alone in DFCI315 and DFCI359. The combination of neratinib and trastuzumab in vivo was more effective compared with single-agent neratinib or trastuzumab and was associated with more robust inhibition of HER2 and downstream signaling.
Conclusions:
The XDOTS platform can be used to evaluate therapies and therapeutic combinations ex vivo using PDX tumors. This approach may accelerate the identification and clinical development of therapies for targets with no or few existing models and/or therapies.
Insights
This study evaluated neratinib drug combinations in HER2-mutant non-small cell lung cancer (NSCLC) using patient-derived xenograft-derived organotypic spheroids (XDOTS). Neratinib combined with trastuzumab or temsirolimus showed promise for treating HER2-mutant NSCLC.
Area of Science:
- Oncology
- Pharmacology
- Biotechnology
Background:
- Primary patient-derived cells offer a rapid ex vivo approach for drug response evaluation.
- HER2-mutant non-small cell lung cancer (NSCLC) presents a challenge for targeted therapy development.
Purpose of the Study:
- To identify effective neratinib combinations for HER2-mutant NSCLC using a novel ex vivo system.
- To evaluate the efficacy of drug combinations in patient-derived xenograft-derived organotypic spheroids (XDOTS).
Main Methods:
- Generated two HER2-mutant NSCLC patient-derived xenograft (PDX) models.
- Established XDOTS from PDX tumors and treated them ex vivo with neratinib-based combinations.
- Validated the most effective ex vivo combination in vivo using PDX models and a genetically engineered mouse model.
Main Results:
- Neratinib and afatinib induced cell death in one HER2-mutant NSCLC model, while gefitinib did not.
- Combinations of neratinib/trastuzumab and neratinib/temsirolimus enhanced neratinib's therapeutic benefit.
- The neratinib/trastuzumab combination demonstrated superior in vivo efficacy and HER2 signaling inhibition compared to single agents.
Conclusions:
- The XDOTS platform is effective for evaluating ex vivo therapies and drug combinations using PDX tumors.
- This approach can accelerate the development of therapies for targets with limited existing models.
- XDOTS facilitates rapid screening of therapeutic strategies for HER2-mutant NSCLC.
