Abstract Introduction: MYC and BCL2 co-expression defines double-expressor lymphoma (DEL), a subtype of diffuse large B-cell lymphoma (DLBCL) associated with poor prognosis. However, its clinical utility is limited by inconsistent scoring thresholds and a possible protective role of BCL6. Using multiplexed fluorescent IHC (mfIHC), we previously showed (Hoppe et al. , Cancer Discovery 2023) that survival is better predicted by the fraction of tumor cells co-expressing MYC and BCL2 in the absence of BCL6 (M+2+6−), refining DEL classification. Methods and Results: In this follow-up study, we assessed the spatial distribution of M+2+6− cells across four independent DLBCL cohorts (n = 449). Geyer’s point process analysis revealed two spatial phenotypes—Clustered and Dispersed. Dispersed M+2+6− cases consistently showed inferior survival (P 0. 05 in 4/4 cohorts), representing the first evidence that spatial architecture of oncogenic subpopulations impacts prognosis. A transcriptional signature derived from dispersed tumors was evaluated in six independent gene expression profiling datasets (n = 4, 594), where dispersed-like profiles also correlated with poor survival (P 0. 05 in 4/6 cohorts). We further examined the tumor microenvironment in 152 cases using hyperplex IHC. Dispersed tumors displayed an immune-cold milieu with enriched regulatory and exhausted T cells, higher expression of IDO1 and LAG3, and strong association with the aggressive activated B-cell (ABC) subtype—suggesting that spatial patterning contributes to immune escape. To enable clinical translation, we developed the scDEL Calculator, a machine-learning–based web tool that estimates 2-year relapse risk using routine IHC scores for MYC, BCL2, and BCL6, and calculates the probability of oncogenic co-expression. Conclusion: This is the first report, to our knowledge, that the spatial pattern of a tumor subpopulation influences survival and immune microenvironment in cancer. The scDEL Calculator (https: //scdel-calc. streamlit. app/) offers a step toward integrating these insights into risk stratification in DLBCL. Citation Format: Shruti Sridhar, Michal Marek Hoppe, Min Liu, Gayatri Kumar, Siddham Jasoria, Aditi Singh, Kanav Gupta, Ziwei Meng, Chartsiam Tipgomut, Patrick Jaynes, Yanfen Peng, Limei Poon, Yen Lin Chee, Susan Swee-Shan Hue, Siok-Bian Ng, Chandramouli Nagarajan, Ong Choon Kiat, Lim Soon Thye, David W. Scott, Xiaofei Ye, Qiang Pan-Hammarström, Vaibhav Rajan, Kasthuri Kannan, Claudio Tripodo, Anand D. Jeyasekharan. The Spatial Organization of Lymphoma Cells with MYC-BCL2 Co-Expression Shapes the Immune Microenvironment and Determines Survival After Chemoimmunotherapy abstract. In: Proceedings of Frontiers in Cancer Science 2025; 2025 Nov 5-7; Singapore. Philadelphia (PA): AACR; Cancer Res 2026;86 (13Suppl): Abstract nr P73.
Sridhar et al. (Thu,) studied this question.