Abstract CT154: B-cell infiltration in lung cancer predicts response to neoadjuvant pembrolizumab
Authors: Bar Jair , Kamer Iris , Zadok Oranit , Urban Damien , Perelman Marina , Redinsky Ilanit , Ackerstein Aliza , Daher Sameh , Ofek Efrat , Onn Amir , Zeitlin Nona , Ben-Nun Alon , Kremer Ran , Daniel Inbal , Glantzspiegel Yossef , Gat-Viks Irit 
Published: June 2022

Abstract
Background Neoadjuvant immune checkpoint inhibitor treatment is a promising approach for resectable cancer, including non-small cell lung cancer (NSCLC). The characteristics of potential responders to such treatments and the molecular underlying events are not known. Methods We have conducted a phase I, investigator-initiated single-center study (MK3457-223), to examine the safety of neoadjuvant pembrolizumab for stage I-II resectable NSCLC and to determine the recommended phase II dose/schedule (RP2D/S). FFPE biopsies and surgical specimens were subjected to correlative studies. NanoString’s GeoMx Digital Spatial Profiler (DSP) analysis was conducted on pre-treatment samples and post-treatment responder samples. Protein (72 proteins) and mRNA expression data (73 genes) analysis was conducted on regions of interest (ROIs), defined as mostly CD8 positive or mostly pan-cytokeratin positive (presumed cancer cells). Pathology assessment was done on the surgical specimen to identify major pathologic response (MPR; ≤10% remaining viable cells). Statistical analysis was done to compare responders (MPR+) to non-responders (MPR-) by Mann Whitney with false discovery rate correction. Immunohistochemistry (IHC) was conducted on post-treatment samples. Results Twenty-six patients initiated treatment on the study. Two patients (8%, 95% C.I 0-18%) had adverse events that precluded surgery, 1 patient refused surgery after treatment. 7 patients (27%, 95% C.I 10-44%) achieved a major pathologic response (MPR; responders), 3 patients (12%, 95% C.I 0-24%) achieved complete pathologic response. Responders had a longer interval from treatment to surgery (43 days vs. 36 days, univariate analysis, p-value 0.043). RP2D/S was determined as 2 treatments of 200mg pembrolizumab at 3 week interval, followed by surgery at least 2 weeks later. The expression of several proteins and genes differed between responders and non-responders. Pre-treatment, CD20 protein was the most differentially expressed protein both in in CD8+ (4.7 fold, p=0.002) and in cancer cells (4.8 fold, p=0.001) ROIs, in both cases higher in the responders compared to the non-responders. Comparing pre to post-treatment expression in responding tumors, the protein found to be upregulated to the highest extent following pembrolizumab treatment was CD20 protein (6.2-fold, p=0.001), as was its encoding gene, MS4A1 (2.4-fold, p=0.006). CD20 IHC of post-treatment samples demonstrated tertiary lymphoid structures (TLS) to be more prevalent in responders compared to non-responders (3.2-fold, p&lt;0.05). Conclusions Longer interval from treatment to surgery was associated with higher rate of MPR. Presence of tumor-infiltrating B-cells and evolvement of TLSs was strongly correlated with pathologic response to neoadjuvant pembrolizumab in early stage NSCLC.
Citation Format: Jair Bar, Iris Kamer, Oranit Zadok, Damien Urban, Marina Perelman, Ilanit Redinsky, Aliza Ackerstein, Sameh Daher, Efrat Ofek, Amir Onn, Nona Zeitlin, Alon Ben-Nun, Ran Kremer, Inbal Daniel, Yossef Glantzspiegel, Irit Gat-Viks. B-cell infiltration in lung cancer predicts response to neoadjuvant pembrolizumab [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 CT154.