Abstract 2023: Multiplex immunofluorescence profiling of the tumor microenvironment and CD73: Activity of neoadjuvant oleclumab in patients with non-small-cell lung cancer in NeoCOAST
Authors: Bisha Ina , Tan Tze Heng , Weitkunat Manuela , Andoni Alma , Dino Iris , Martin Philip , Saraiya Megha , DaCosta Karma , Sontakke Pallavi , Schick Markus , Surace Michael , Blando Jorge , Grenga Italia , Kumar Rakesh , McGrath Lara 
Published: March 2024

Abstract
Background: The NeoCOAST study (NCT03794544) investigated the efficacy of neoadjuvant immuno-oncology combinations in patients with resectable non-small-cell lung cancer (NSCLC), using major pathological response (MPR) as the primary endpoint (Cascone T, et al. Cancer Discov 2023;13:2394-411). Patients received a single cycle of the following treatments: durvalumab monotherapy (durva; anti-PD-L1), durva + oleclumab (ole; anti-CD73), or durva + monalizumab (mona; anti-NKG2A). Here we analyze the tumor microenvironment to investigate the mechanism of action of ole.
Methods: A cohort of 47 patients (17 durva, 13 durva + mona, 17 durva + ole) with tumor tissue samples from pre-treatment (baseline, n=31) and surgery (post-treatment, n=24; n=8 paired pre- and post-treatment samples) were evaluated. Tumor sections were stained with (1) multiplex immunofluorescence (Panel 1: CD8-PD-L1-PD1-CD68-Ki67-panCK, Panel 2: NKp46-CD20), (2) CD73 immunohistochemistry (IHC), and (3) NKG2A IHC. Stains were analyzed using deep learning algorithms, manual pathology, and digital pathology scoring, respectively.
Results: All arms showed similar infiltration of tumors by T cells (CD8), B cells (CD20), NK cells (NKp46), and macrophages (CD68) at the pre-treatment time point (Kruskal-Wallis, p=0.5-0.9 n.s.). Percent CD73+ tumor cells (TCs) at baseline was correlated with increased B cell, T cell, NK cell, and macrophage abundance (Spearman’s Rho=0.4-0.6, p&lt;0.05). Percent CD73+ TC and B cell abundance was higher in patients with MPR vs those without MPR in the durva + ole arm, but not in the durva monotherapy or durva + mona arms. Increased CD8 T cell and NK cell abundance was not associated with MPR in any arm. Moreover, comparison of baseline to post-treatment tumor samples revealed a greater increase in CD8 T cells in the durva + ole arm (FC=2.1, p=0.02), particularly in proliferating activated T cells (CD8+Ki67+PD-1+; FC=8.0, p=0.003), compared to durva alone (FC=1.8, p=0.4 and FC=2.9, p=0.04, respectively).
Conclusion: Increased abundance of CD73+ TCs is correlated with high B cells in the tumor microenvironment and associated with patients who experienced MPR in the durva + ole arm. Treatment with neoadjuvant durva + ole results in a greater increase in proliferating CD8 T cells than durva alone.
Citation Format: Ina Bisha, Tze Heng Tan, Manuela Weitkunat, Alma Andoni, Iris Dino, Philip Martin, Megha Saraiya, Karma DaCosta, Pallavi Sontakke, Markus Schick, Michael Surace, Jorge Blando, Italia Grenga, Rakesh Kumar, Lara McGrath. Multiplex immunofluorescence profiling of the tumor microenvironment and CD73: Activity of neoadjuvant oleclumab in patients with non-small-cell lung cancer in NeoCOAST [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2023.