Abstract 4460: Spatial profiling of immune biomarkers in resected treatment-naïve early stage lung adenocarcinoma
Authors: Hernandez Sharia D. , Lu Wei , Serrano Alejandra G. , Arrechedera Claudio J. , Sanchez-Espiridion Beatriz , Ozirmak Nejla , Molina Max , Kostousov Larisa , Barnes Sean , Khan Khaja , Tang Ximing , Fujimoto Junya , Parra Edwin R. , Raso Gabriela , Schmidt Stephanie T. , Behrens Carmen , Heymach John , Zhang Jianjun , Chen Ken , Sepesi Boris , Cascone Tina , Gibbons Don , Wistuba Ignacio I. , Haymaker Cara , Solis Luisa M. 
Published: April 2023

Abstract
Background: Recently, neoadjuvant immunotherapy plus chemotherapy has been approved for treatment of resectable non-small cell lung carcinoma (NSCLC). Defining the immune landscape of these tumors and its spatial distribution will help to understand lung cancer biology. Here, we analyzed the distribution of immune-related biomarkers in tumor-defined regions and its associations with clinicopathological variables in resected lung adenocarcinomas using high-plex profiling approaches.
Methodology: Thirty-three FFPE tumor tissues from surgically resected treatment-naïve lung adenocarcinoma stage I/II were used to construct a tissue microarray from the MD Anderson ICON cohort. We used three 1-mm core per patient [2 from central tumor (CT), and 1 from invasive margin (IM)] and performed the GeoMx Digital Spatial Profiling protein protocol to assess 49 immune biomarkers. Pancytokeratin (panCK; epithelial), CD45 (immune) and SYTO 13 (nuclear) were utilized as morphology biomarkers. Regions of interests were placed in cores containing tumor, and segmented in ‘”Tumor (Tu)” (PanCK+) and the “tumor microenvironment (TME)” (PanCK-). Digital counts were normalized using background correction. Statistical analysis was performed using linear mixed model. A p value equal or less than 0.05 was considered significant.
Results: We first compared the relative counts of immune biomarkers in the TME in CT and IM. IM had higher CD3, CD8, CD45RO, as well as CD163 and STING (P ranges 0.006 to 0.035), while CT had higher PD1 (P 0.025). Then we analyzed differential biomarker expression by sex and smoking status. Females had higher counts of Immune related biomarkers: CD45, CD3, CD20, immune checkpoints: PD-L1, VISTA, CTLA4, LAG3, and ICOS, and myeloid: CD68, CD11c, CD163, and B2M (P ranges 0.0005 to 0.046). Smokers had higher counts of CD66b (P 0.007) and B2M (P 0.039) while never smokers had higher counts of HLA-DR, CD34, FoxP3, OX40L, Tim-3, and B7-H3 (P ranges 0.001 to 0.049). Finally, we analyzed Tu segments. IM had higher CD66b, VISTA, CD163, OX40L, HLA-DR, GZMB, STING, and CD8 (P ranges 0.007 to 0.045) than CT. Female patients had higher CD45, CD68, CD11c and CD163 (P ranges 0.008 to 0.045), and males had higher SMA (P 0.005). Smokers had higher CD66b (P 0.007), B2-microglobulin (P 0.043), and never smokers had higher HLA-DR, STING, CD34, CD44, FoxP3 and CD25 (P ranges 0.006 to 0.043).
Conclusions: In this study, biomarker analysis of treatment-naïve adenocarcinoma in CT areas and IM indicates a higher immune response in the IM and presence of inhibitory signaling inside the tumor. Our data also showed that tumors from females have higher immune response than tumors from males, which is concordant with previous studies. Distinct profiling by smoking status was also observed. Further analysis of gene expression analysis of this set is ongoing.
Citation Format: Sharia D. Hernandez, Wei Lu, Alejandra G. Serrano, Claudio J. Arrechedera, Beatriz Sanchez-Espiridion, Nejla Ozirmak, Max Molina, Larisa Kostousov, Sean Barnes, Khaja Khan, Ximing Tang, Junya Fujimoto, Edwin R. Parra, Gabriela Raso, Stephanie T. Schmidt, Carmen Behrens, John Heymach, Jianjun Zhang, Ken Chen, Boris Sepesi, Tina Cascone, Don Gibbons, Ignacio I. Wistuba, Cara Haymaker, Luisa M. Solis. Spatial profiling of immune biomarkers in resected treatment-naïve early stage lung adenocarcinoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4460.