Abstract 1116: Spatial distribution of granzyme B to improve the predictive power of response to immunotherapy in operable lung cancer
Authors: Ramos Corinne , Lemieux Ophelie , Ponzoni Adele , Stauber Jonathan 
Published: June 2022

Abstract
Purpose Non-small cell lung cancer (NSCLC) accounts for about 85% of all lung cancers. There is a strong rationale for incorporating immunotherapy into the treatment of early-stage NSCLC, given the breakthrough results with PD-1 checkpoint inhibitors in advanced-stage NSCLC. How immunotherapy should be implemented in patients who are operable is still unclear. Most of the efforts so far to identify clinically useful biomarkers do not preserve spatial information and leave us blind to the critical source of information revealed in the cell-to-cell biology of the tumor microenvironment (TME). In order to overcome these limitations, we used spatial biomarkers assays that preserve this critical information about which cells are influencing treatment response.
Method Frozen sections from retrospectively collected surgically resected NSCLC (adenocarcinoma and squamous cell carcinoma) tumors treated with adjuvant pembrolizumab therapy were used. Patients were classified in two groups according to their Objective Response Rate (ORR): Complete Response (CR) and Progression Disease (PD) for spatial transcriptomic and proteomics assays. Associated to RNA In situ hybridization (ISH) technology, the NanoString GeoMx™ Digital Spatial Profiling (DSP) technology was used to determine immune markers within compartment specific defined by fluorescence localization [tumor (panCK) and leucocytes (CD45)]. Wilcoxon, Linear Mixed Model and Mann-Whitney U statistic tests were used to evaluate differences between ORR groups.
Results Using the spatial analysis, we identified 4 protein markers from immune cell profiling and immune activation status modules independently associated with benefit from single-agent PD-1 checkpoint blockade in spatial context. High expression of CD4, CD40, Granzyme B and HLA-DR were significantly associated with all favorable clinical outcome, whereas upregulation of extracellular matrix (ECM) proteins related to pro-tumoral reprogramming of the stroma were associated with immunotherapy resistance. We also validated the DSP finding that high granzyme level in the stroma compartment was predictive for response to check point inhibitor in the same set of patients using ISH immunofluorescence, strengthening this marker relevance in antitumor immunity and as a predictive biomarker candidate for immunotherapy in NSCLC.
Conclusion This work identifies a number of relevant spatial candidate predictors of immunotherapy outcome in spatial context for operable lung cancer that show promise for future validation in larger independent cohorts.
Citation Format: Corinne Ramos, Ophelie Lemieux, Adele Ponzoni, Jonathan Stauber. Spatial distribution of granzyme B to improve the predictive power of response to immunotherapy in operable lung cancer [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 1116.