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Updated: Jun 25, 2025
Multiparametric Tumor Organoid Drug Screening Using Widefield Live-Cell Imaging for Bulk and Single-Organoid Analysis
Published on: December 23, 2022
OrganoIDNet: a deep learning tool for identification of therapeutic effects in PDAC organoid-PBMC co-cultures from
Nathalia Ferreira1, Ajinkya Kulkarni1, David Agorku2,3
1Translational Molecular Imaging, Max-Planck-Institute for Multidisciplinary Sciences, Göttingen, Germany.
Purpose:
Pancreatic Ductal Adenocarcinoma (PDAC) remains a challenging disease due to its complex biology and aggressive behavior with an urgent need for efficient therapeutic strategies. To assess therapy response, pre-clinical PDAC organoid-based models in combination with accurate real-time monitoring are required.
Methods:
We established stable live-imaging organoid/peripheral blood mononuclear cells (PBMCs) co-cultures and introduced OrganoIDNet, a deep-learning-based algorithm, capable of analyzing bright-field images of murine and human patient-derived PDAC organoids acquired with live-cell imaging. We investigated the response to the chemotherapy gemcitabine in PDAC organoids and the PD-L1 inhibitor Atezolizumab, cultured with or without HLA-matched PBMCs over time. Results obtained with OrganoIDNet were validated with the endpoint proliferation assay CellTiter-Glo.
Results:
Live cell imaging in combination with OrganoIDNet accurately detected size-specific drug responses of organoids to gemcitabine over time, showing that large organoids were more prone to cytotoxic effects. This approach also allowed distinguishing between healthy and unhealthy status and measuring eccentricity as organoids' reaction to therapy. Furthermore, imaging of a new organoids/PBMCs sandwich-based co-culture enabled longitudinal analysis of organoid responses to Atezolizumab, showing an increased potency of PBMCs tumor-killing in an organoid-individual manner when Atezolizumab was added.
Conclusion:
Optimized PDAC organoid imaging analyzed by OrganoIDNet represents a platform capable of accurately detecting organoid responses to standard PDAC chemotherapy over time. Moreover, organoid/immune cell co-cultures allow monitoring of organoid responses to immunotherapy, offering dynamic insights into treatment behavior within a co-culture setting with PBMCs. This setup holds promise for real-time assessment of immunotherapeutic effects in individual patient-derived PDAC organoids.
Insights
This study introduces OrganoIDNet, a deep-learning tool for real-time monitoring of pancreatic cancer organoid responses to chemotherapy and immunotherapy. It enables personalized treatment assessment for pancreatic ductal adenocarcinoma (PDAC).
Area of Science:
- Oncology
- Biotechnology
- Artificial Intelligence
Background:
- Pancreatic Ductal Adenocarcinoma (PDAC) is a challenging cancer with limited therapeutic options.
- Efficient therapeutic strategies and accurate response assessment are crucial for PDAC.
- Pre-clinical organoid models are essential for studying PDAC and evaluating new treatments.
Purpose of the Study:
- To develop and validate a real-time monitoring system for PDAC organoid response to therapies.
- To assess the efficacy of chemotherapy (gemcitabine) and immunotherapy (Atezolizumab) using organoid models.
- To investigate the role of immune cells (PBMCs) in PDAC organoid response to immunotherapy.
Main Methods:
- Established live-imaging co-cultures of PDAC organoids and peripheral blood mononuclear cells (PBMCs).
- Developed OrganoIDNet, a deep-learning algorithm for analyzing bright-field images of organoids.
- Investigated responses to gemcitabine and Atezolizumab, with and without PBMCs, using live-cell imaging and validated with CellTiter-Glo.
Main Results:
- OrganoIDNet accurately detected size-specific gemcitabine responses in PDAC organoids over time.
- The system distinguished organoid health status and measured therapy-induced eccentricity.
- Organoid/PBMC co-cultures revealed enhanced Atezolizumab potency in PBMC-mediated tumor killing.
Conclusions:
- Optimized PDAC organoid imaging with OrganoIDNet accurately detects chemotherapy responses.
- Organoid/immune cell co-cultures enable real-time monitoring of immunotherapy effects.
- This platform offers dynamic insights for personalized assessment of PDAC treatment.
