Abstract 2548: The central nervous system immune cell interactome is a function of cancer lineage, tumor microenvironment and STAT3 expression
Authors: Najem Hinda , Ott Martina , Kassab Cynthia , Rao Arvind , Rao Ganesh , Marisetty Anantha , Sonabend Adam M. , Horbinski Craig , Verhaak Roel , Shakar Anand , Krishnan Santhoshi , Varn Frederick S. , Arietta Victor A. , Gupta Pravesh , Ferguson Sherise D. , Huse Jason , Fuller Gregory N. , Long James , Winskowski Dan , Freiberg Ben , James C. David , Platanias Leonidas C. , Lesniak Maciej S. , Burks Jared K. , Heimberger Amy B. 
Published: June 2022

Abstract
Introduction: Deconstructive immune cell profiling of central nervous system (CNS) tumors has focused on the tumor, excluding interrogation of the tumor microenvironment (TME). Integrated spatial analysis can ascertain the cell interactome and may be a key biomarker for effective anti-tumor immune responses.
Methods: En bloc resections of glioma (n=10) and lung metastasis (n=10) to preserve the tissue architecture, underwent tissue segmentation and high dimension opal 7-color multiplex imaging. Bioinformatic analysis of scRNA was used to infer immune cell functionality.
Results: CD3+ T cell frequency was equivalent between CNS cancer lineages. Within gliomas T cells were confined to the perivascular space and the infiltrating edge. In lung metastasis, T cells are confined to the tumor stroma. CD163+ macrophages predominate in brain metastasis throughout the TME (p&lt;0.05), while CD68+ monocytes (CD68+, CD11c+CD68+, and CD11+CD68+CD163+) are more common in gliomas (p&lt;0.05). T cell dyad and cluster immune interactions were more common in the absence of nuclear STAT3 expression. T cells usually interact with CD163+ macrophages as dyads in metastasis at the brain interface (p=0.031) and within tumor (p=0.0009); in clusters throughout the TME (interface: p=0.024; tumor: p=0.01; necrosis: p=0.045), and as STAT3+ dyads and cluster interactions in the tumor (p&lt;0.05). Immune suppressed CD11c+CD163+ dendritic cells (tumor: p=0.036; and necrosis p=0.020) predominate in metastasis. In contrast, gliomas typically lacked dyad and cluster interactions except for T cell and CD68+ dyads in the tumor (p=0.023). Bioinformatic analysis of CD45+ scRNA seq data revealed that the majority of innate immune populations express both pro-inflammatory and immune suppressive genes and that subsets of CD68+ and CD11c+CD68+ cells expressed markers such as TMEM119, P2YR13 and CX3CR1 that identify microglia.
Conclusion: Current therapies are targeted to cell populations and singular pathways. Immunosuppressive macrophages dominate within the TME and targeting this population may create an environment that favors T cell activation and effective immune responses. Furthermore, the immune interactome, an important event for anti-tumor immune response, is a function of cancer lineage, TME, and STAT3 expression, which will gain relevance for future therapeutics directed to modulating these interactions.
Citation Format: Hinda Najem, Martina Ott, Cynthia Kassab, Arvind Rao, Ganesh Rao, Anantha Marisetty, Adam M. Sonabend, Craig Horbinski, Roel Verhaak, Anand Shakar, Santhoshi Krishnan, Frederick S. Varn, Victor A. Arietta, Pravesh Gupta, Sherise D. Ferguson, Jason Huse, Gregory N. Fuller, James Long, Dan Winskowski, Ben Freiberg, C. David James, Leonidas C. Platanias, Maciej S. Lesniak, Jared K. Burks, Amy B. Heimberger. The central nervous system immune cell interactome is a function of cancer lineage, tumor microenvironment and STAT3 expression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2548.