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bioRxiv · 10.1101/2025.09.04.674374

Systematic Growth Factor Profiling Platform for 3D Tumor Models Reveals Estradiol-Responsive Cellular Mechanisms of Immunotherapy Resistance

Lee, J.-K.·Lee, K.·Kim, M.·Lim, S. O.·Shin, D.-J.·Shin, Y.·Choi, J.-J.·Lee, M.·Kang, H. J.·Lee, J.-W.
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Abstract

Current organoid culture systems face critical limitations: standardized growth factor formulations fail to capture patient-specific signaling requirements, and single-cell-type approaches overlook tumor-stromal interactions essential for understanding immunotherapy resistance. To address these challenges, we developed an innovative biofabrication platform that systematically integrates patient-derived three-dimensional (3D) cultures with comprehensive growth factor profiling across 128 combinations. Through systematic screening of 23 ovarian cancer patient samples and single-cell RNA sequencing, we identified two estradiol-responsive cellular populations that coordinate immunosuppression: a malignant cell fraction (MAL.PDCD5) and a cancer-associated fibroblast (CAF) fraction (FB.TNFSF10). MAL.PDCD5 cells suppress immune infiltration by downregulating antigen presentation, while FB.TNFSF10 cells promote immunosuppression through enhanced TGF-{beta} signaling. Spatial transcriptomic analysis revealed striking mutual exclusivity between FB.TNFSF10 cells and T/NK cells, providing evidence of active immune cell exclusion. Most significantly, FB.TNFSF10 abundance emerged as a robust predictor of immune checkpoint inhibitor therapy resistance across multiple cancer cohorts, independent of conventional biomarkers. This biofabrication platform provides a scalable framework for drug screening and biomarker discovery, with immediate applications in precision medicine for patient stratification and combination therapy development.

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Lee, J.-K., Lee, K., Kim, M., Lim, S. O., Shin, D.-J., Shin, Y., Choi, J.-J., Lee, M., Kang, H. J., Lee, J.-W.. 2025-09-09. Systematic Growth Factor Profiling Platform for 3D Tumor Models Reveals Estradiol-Responsive Cellular Mechanisms of Immunotherapy Resistance. https://doi.org/10.1101/2025.09.04.674374

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