{
  "abstract": "Background Double-expressor lymphomas (DEL) overexpressing MYC and BCL2 show poor outcomes in diffuse large B-cell lymphoma (DLBCL). We previously demonstrated that survival correlates with malignant cells co-expressing MYC+BCL2+ while lacking BCL6 (M+2+6- cells). Here, we investigate whether spatial organization of M+2+6- cells influences clinical outcomes and tumor microenvironment composition.Methods Using Geyer’s point process analysis on multiplexed immunohistochemistry data, we classified M+2+6- cell spatial patterns as ”clustered” or ”dispersed” across 449 patients in four independent cohorts. Immune microenvironments were characterized using de novo DLBCL single-cell RNA-seq (n=17), WTA Digital Spatial Profiling (n=96), and a 58-marker hyperplex PhenoCycler analysis (n=152). Lymphoma Microenvironment Archetype Profiles (LymphoMAPs) were analyzed in a single cell RNASeq cohort of 92 r/r DLBCL samples.Results Dispersed M+2+6- cell distribution significantly predicted shorter survival across all cohorts (P<0.05), representing the first demonstration that spatial tumor cell organization impacts clinical outcomes in lymphoma. Dispersed cases showed enhanced M+2+6- T cell interactions by scRNA-seq and were enriched for regulatory T cell and exhausted T cell signatures by DSP. Hyperplex analysis confirmed higher FOXP3+ Treg proportions (p=0.027) and closer spatial proximity between dispersed M+2+6- cells and Tregs. PhenoCycler analysis revealed elevated IDO1 and LAG3 expression (p<0.05) with enrichment of IDO1+ dendritic cells and LAG3+ CD8 T cells in dispersed cases. LymphoMAP analysis showed 44% of dispersed cases belonged to the immunosuppressive TEX (T cell exclusion) archetype versus minimal LN representation.Conclusions Spatial organization of M+2+6- cells is a novel facet in DLBCL. Dispersed patterns create immunosuppressive microenvironments promoting IDO1+ dendritic cells, LAG3+ exhausted T cells, and Treg infiltration. This could enhance risk stratification and identify patients who may benefit from CAR T therapies.",
  "authors": [
    {
      "affiliations": [
        "National University of Singapore, Singapore, Singapore"
      ],
      "name": "Shruti Sridhar"
    },
    {
      "affiliations": [
        "National University of Singapore, Singapore, Singapore"
      ],
      "name": "Charmaine Ong"
    },
    {
      "affiliations": [
        "National University of Singapore, Singapore, Singapore"
      ],
      "name": "Irene Biju"
    },
    {
      "affiliations": [
        "The University of Texas MD Anderson Cancer Center, Houston, TX, USA"
      ],
      "name": "Xubin Li"
    },
    {
      "affiliations": [
        "National University of Singapore, Cancer Science Institute of Singapore, Singapore, Singapore"
      ],
      "name": "Michal M Hoppe"
    },
    {
      "affiliations": [
        "National University of Singapore, Department of Pathology, Yong Loo Lin School of Medicine, Singapore, Singapore"
      ],
      "name": "Shuangyi Fan"
    },
    {
      "affiliations": [
        "National University of Singapore, Department of Haematology-Oncology, Singapore, Singapore"
      ],
      "name": "Sanjay de Mel"
    },
    {
      "affiliations": [
        "National University of Singapore, Singapore, Singapore"
      ],
      "name": "Limei Poon"
    },
    {
      "affiliations": [
        "National University of Singapore, Singapore, Singapore"
      ],
      "name": "Kiat Ong"
    },
    {
      "affiliations": [
        "National Cancer Centre, Singapore, Singapore"
      ],
      "name": "Soon Thye Lim"
    },
    {
      "affiliations": [
        "National Cancer Centre, Singapore, Singapore"
      ],
      "name": "Chandramouli Nagarajan"
    },
    {
      "affiliations": [
        "National Cancer Centre, Singapore, Singapore"
      ],
      "name": "Nicholas Grigoropoulos"
    },
    {
      "affiliations": [
        "BC Cancer Research Centre, Vancouver, BC, Canada"
      ],
      "name": "David Scott"
    },
    {
      "affiliations": [
        "National University of Singapore, Singapore, Singapore"
      ],
      "name": "Wee Joo Chng"
    },
    {
      "affiliations": [
        "National University of Singapore, Department of Pathology, Yong Loo Lin School of Medicine, Singapore, Singapore"
      ],
      "name": "Siok Bian Bian Ng"
    },
    {
      "affiliations": [
        "The University of Texas MD Anderson Cancer Center, Houston, TX, USA"
      ],
      "name": "Michael Green"
    },
    {
      "affiliations": [
        "University of Palermo, Tumor Immunology Unit, Palermo, PA, Italy"
      ],
      "name": "Claudio Tripodo"
    },
    {
      "affiliations": [
        "National University of Singapore, Singapore, Singapore"
      ],
      "name": "Anand Jeyesekharan"
    }
  ],
  "title": "1285 Spatial organization of MYC+BCL2+BCL6- cells predicts survival and defines immunosuppressive microenvironments in diffuse large B-cell lymphoma",
  "uid": "feb0d0a7-5391-5231-a28d-7579b3fa7d9f"
}