FIGURE 3 from Neoadjuvant CD40 Agonism Remodels the Tumor Immune Microenvironment in Locally Advanced Esophageal/Gastroesophageal Junction Cancer

• DOI: 10.1158/2767-9764.25061895.v1
• Publication Date: 2024-01-25T14:26:10Z
• Authors: Maira Soto; Erin L. Filbert; Hai Yang; Stephanie Starzinski; Alec Starzinski; Marissa Gin; Brandon Chen; Phi Le; Tony Li; Brandon Bol; Alexander Cheung; Li Zhang; Frank J. Hsu; Andrew Ko; Lawrence Fong; Bridget P. Keenan

<p>Sotigalimab induces an activated T cell infiltrate and reduces Tregs in tumors. <b>A,</b> Characterization of T cell subsets in the TME was done by MIBI analysis using markers for DNA, keratin, CD4, CD8, CD45RO, Foxp3, and Granzyme B (GZMB; representative image: patient 04). <b>B,</b> Percentage of each T cell subsets are shown for samples pre- and post-treatment, out of total intratumoral T cells, as analyzed by MIBI (pre <i>n</i> = 5, post <i>n</i> = 4). <b>C,</b> Ki67 expression in all CD8⁺ T cell subsets was quantified using MIBI analysis (pre <i>n</i> = 5, post <i>n</i> = 4). <b>D,</b> scRNAseq analysis was used to identify the top pathways either up- or downregulated in T cells (pre <i>n</i> = 3, post <i>n</i> = 4). The text inside boxes of the heat map is the proportion of genes enriched in the pathway; gray color indicates no enriched genes. <b>E,</b> Individual cells were scored for genes related to oxidative phosphorylation (<i>y</i>-axis) and T cell effector function (<i>x</i>-axis) for CD8⁺ and non-Treg CD4⁺ T cells (pre <i>n</i> = 3, post <i>n</i> = 4). <b>F,</b> Quantification of T cell clones as preexisting, newly induced, or persistent for each T cell type in the tumor (pre <i>n</i> = 3, post <i>n</i> = 4). <b>G</b> and <b>H,</b> More granular subtypes of T cells plotted in UMAP space (G) and labeled by subtype, and quantified to show distribution of preexisting, induced, or persistent clones (H). <b>I,</b> Network plot of T cell clones (paired alpha and beta chain) with cluster details (newly induced, persistent, or pre-existing clones; blood or tumor compartment; T cell type) or patient identity overlaid (tumor: pre <i>n</i> = 3, post <i>n</i> = 4; blood: pre <i>n</i> = 6, post <i>n</i> = 6). All statistical changes for MIBI data were measured by <i>t</i> test using Prism GraphPad. *, <i>P</i> ≤ 0.05; **, <i>P</i> ≤ 0.01; ***, <i>P</i> ≤ 0.001. For scRNAseq data, comparisons denoted as significant have adjusted <i>P</i> < 0.05.</p>