Abstract 6391: Spatial proteomic profiling unveils pre-existing anti-tumor immunity as a hallmark of exceptional benefit from immunotherapy in NSCLC
Authors: Hernandez Sharia , Bach Rafael , Giner Mario , Lu Wei , Kostousov Larisa , Barnes Sean , Khan Khaja , Masfarré Laura , Villanueva Xavier , Sanchéz Ignacio , Navarro Nil , Taus Álvaro , Galindo Miguel , Hardy Max , Rey-Vergara Raúl Del , Iñañez Albert , Sanchez-Espiridion Beatriz , Moliner Laura , Clavé Sergi , Bellosillo Beatriz , Rovira Ana , Perera-Bel Júlia , Wistuba Ignacio , Arriola Edurne , Solis Luisa M. , Rocha Pedro 
Published: March 2024

Abstract
Background: Immunotherapy is firmly established as a treatment regimen in various solid tumors due to its exceptional benefits observed in a select group of patients. Despite widespread use of immune checkpoint blockade (ICB) across diverse solid tumors, including non-small cell lung cancer (NSCLC), the quest for a clinically informative biomarker for long-term benefit remains unmet. Here we investigate the potential utility of clinicopathological and spatial proteomic profiling of tumor specimens from NSCLC patients to identify predictive biomarkers of long-term benefit to ICB.
Methods: Forty-nine patients diagnosed with advanced NSCLC who received ICB at Hospital del Mar, Barcelona between 2017-2021 were included. Long-term responders (LTR, n=21) were defined as patients achieving a maintained radiologic response for more than 2 years, and short-term responders (STR, n=28) as patients presenting disease progression within the first six months of ICB initiation. Clinicopathological information, incidence of immune related adverse (irAES) and PD-L1 tumor proportion score assessed by IHC was available for all of them. DSP GeoMx was performed in subset of patients (LTR n=14 and STR n=14), to assess 49 immune biomarkers. Pancytokeratin (panCK; epithelial), CD3, CD20 and SYTO 13 (nuclear) were utilized as morphology biomarkers. Regions of interest were placed in tissue areas containing tumor and segmented in Tumor (PanCK+) and tumor microenvironment (TME) (PanCK-) that was used on downstream analysis. Digital counts were normalized using the protein expression of housekeeper proteins. Statistical analysis was performed using linear mixed models. A P value less than 0.05 was considered significant.
Results: Our analysis revealed specific characteristics of LTR patients compared with STR, namely LTR population was enriched for PD-L1 positive in tumors (p=0.005) and a higher incidence of immune-related adverse events (irAEs) (p=0.001). Within the Tumor compartment, LTR patients displayed significantly higher levels of IDO1, CD8, PD-L1, CD45, HLA-DR and STING, while STR patients exhibit higher levels of B7-H3, CD56 and OX40L (p&lt;0.05 for all proteins). Comparison analysis of the TME compartment between LTR and STR revealed augmented levels of IDO1, CD8, CD45, CD11c, CD27 and CD3 in LTR patients, and lower levels of B7H3 and OX40L compared with STR patients (p&lt;0.05 for all proteins).
Conclusions: Our comprehensive analysis of metastatic NSCLC patients treated with ICB has unveiled distinct clinicopathological and immunological features associated with long-term benefit of ICB, highlighting the presence of pre-existing antitumor immunity as a stronger predictor of long-term benefit. These findings offer insights into potential biomarkers and therapeutic strategies for enhancing ICB outcomes in metastatic NSCLC.
Citation Format: Sharia Hernandez, Rafael Bach, Mario Giner, Wei Lu, Larisa Kostousov, Sean Barnes, Khaja Khan, Laura Masfarré, Xavier Villanueva, Ignacio Sanchéz, Nil Navarro, Álvaro Taus, Miguel Galindo, Max Hardy, Raúl Del Rey-Vergara, Albert Iñañez, Beatriz Sanchez-Espiridion, Laura Moliner, Sergi Clavé, Beatriz Bellosillo, Ana Rovira, Júlia Perera-Bel, Ignacio Wistuba, Edurne Arriola, Luisa M. Solis, Pedro Rocha. Spatial proteomic profiling unveils pre-existing anti-tumor immunity as a hallmark of exceptional benefit from immunotherapy in NSCLC [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 6391.