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Patient-derived organoids (PDOs) can model personalized therapy responses; however, current screening technologies cannot reveal drug response mechanisms or how tumor microenvironment cells alter therapeutic performance. To address this, we developed a highly multiplexed mass cytometry platform to measure post- translational modification (PTM) signaling, DNA damage, cell-cycle activity, and apoptosis in >2,500 colorectal cancer (CRC) PDOs and cancer-associated fibroblasts (CAFs) in response to clinical therapies at single-cell resolution. To compare patient- and microenvironment-specific drug responses in thousands of single-cell da- tasets, we developed ‘‘Trellis’’—a highly scalable, tree-based treatment effect analysis method. Trellis single- cell screening revealed that on-target cell-cycle blockage and DNA-damage drug effects are common, even in chemorefractory PDOs. However, drug-induced apoptosis is rarer, patient-specific, and aligns with cancer cell PTM signaling. We find that CAFs can regulate PDO plasticity—shifting proliferative colonic stem cells (proCSCs) to slow-cycling revival colonic stem cells (revCSCs) to protect cancer cells from chemotherapy.
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This content will become publicly available on April 14, 2026
Targeting CAFs-Mediated Stromal Signaling in a Patient-Derived Organotypic Colorectal Tumor Model
Abstract Colorectal cancer, a significant cause of cancer-related mortality, often exhibits drug resistance, highlighting the need for improved tumor models to advance personalized drug testing and precision therapy. We generated organoids from primary colorectal cancer cells cultured through the conditional reprogramming technique, establishing a framework to perform short-term drug testing studies on patient-derived cells. To model interactions with stromal cells in the tumor microenvironment, we combined cancer cell organoids with carcinoma-associated fibroblasts, a cell type implicated in disease progression and drug resistance. Our organotypic models revealed that carcinoma-associated fibroblasts promote cancer cell proliferation and stemness primarily through hepatocyte growth factor–MET paracrine signaling and activation of cyclin-dependent kinases. Disrupting these tumor–stromal interactions reduced organoid size while limiting oncogenic signals and cancer stemness. Leveraging this tumor model, we identified effective drug combinations targeting colorectal cancer cells and their tumorigenic activities. Our study highlights a path to incorporate patient-derived cells and tumor–stromal interactions into a drug testing workflow that could identify effective therapies for individual patients.
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- Award ID(s):
- 2140091
- PAR ID:
- 10642220
- Publisher / Repository:
- PubMed
- Date Published:
- Journal Name:
- Molecular Cancer Therapeutics
- Volume:
- 24
- Issue:
- 8
- ISSN:
- 1535-7163
- Page Range / eLocation ID:
- 1265 to 1276
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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