Abstract P6-17-08: Dynamics of tumor-infiltrating lymphocytes (TILs) during neoadjuvant dual HER2 blockade in HER2-positive (HER2+) breast cancer in the absence of chemotherapy
Authors: Griguolo G , Holgado E , Cortés J , Fasani R , Pascual T , Paré L , Bermejo B , Oliveira M , Morales S , Martinez N , Vidal M , Pernas S , Lopez R , Muñoz M , Galvan P , Garau I , Manso L , Alarcón J , Martínez E , Villagrasa P , LLombart-Cussac A , Prat A , Nuciforo P 
Published: May 2019

Abstract
Background: TILs in HER2+ breast cancer (BC) predict 1) prognosis in early setting, 2) complete pathological response (pCR) following neoadjuvant antiHER2-based therapy and 3) response to trastuzumab and pembrolizumab in the metastatic setting. However, less is known regarding changes in TILs during antiHER2-based treatment.
Methods: Stromal TILs where evaluated centrally using H/E slides in tumor samples from the PAMELA (NCT01973660) neoadjuvant phase II trial. Briefly, 151 women with HER2+ BC were treated with lapatinib and trastuzumab, and hormonal therapy if HR positive, for 18 weeks. TIL levels were determined at baseline (n=148), after 2 weeks of treatment (n=134) and at surgery (n=137). Expression of 560 genes, including immune-related genes (e.g. CD8A, CD4, PD1 and PDL1) was measured at the same timepoints (baseline n=151, 2-weeks n=144, surgery n=144) using the nCounter platform. Intrinsic subtyping at baseline was determined using the PAM50 gene expression predictor. Changes in TILs between 2 time-points were determined by paired t-tests. Correlation of TILs with gene expression was assessed by quantitative SAM analysis using a False Discovery Rate &lt;1%. All statistical tests were two-sided and considered significant when p&lt;0.05. All statistical analyses were carried out using the R software.
Results: Compared to baseline, a significant increase in TILs was observed at week 2 in HR- (p&lt;0.001) and HER2-enriched (HER2-E) tumors (p=0.001), but not in HR+ (p=0.133) and non-HER2-E tumors (p=0.067). Within HR- and HER2-E tumors, increase in TILs at week 2 from baseline was observed regardless of pathological response at surgery (pCR and HR- [p=0.008]; RD and HR- [p=0.037]; pCR and HER2-E [p=0.010]; RD and HER2-E [p=0.056]). Compared to week 2, a significant decrease in TILs at surgery was observed in HR- (p=0.002) and HER2-E (p=0.003) tumors, but not in HR+ (p=0.616) and non-HER2-E tumors (p=0.578). Within HR- and HER2-E tumors, a significant decrease in TILs between week 2 and surgery was observed in tumors achieving pCR (p=0.004 and p=0.005), while, in tumors not achieving pCR, no significant tendency was observed (26.4% and 33.0% of tumors showed an increase and a decrease of TILs between week 2 and surgery). Nonetheless, the vast majority of residual tumors (non-pCR) at surgery had TILs above ≥5%: 34.3% 5-10%, 21.0% 10-20%, 15.2% 20-40% and 11.4% ≥40%. Finally, TILs scoring was found highly enriched (FDR&lt;1%) for immune-related genes tracking activated CD8 T-cells (i.e. CD8A, CD3G, LAG3 and PD1). Expression of these immune genes consistently correlated with TIL levels across the 3 time-points.
Conclusions: In early HER2+ BC, a general increase in TILs is observed following 2 weeks of dual HER2 blockade. This observation is mostly observed in HR- and HER2-E subtype, but regardless of pathological response at surgery. After 2 weeks of treatment, TILs consistently decrease in patients achieving a pCR, whereas two main patterns of TILs expression are observed in patients with residual disease at surgery. Nonetheless, most residual tumors at surgery are inflamed (i.e. TILs ≥5%) and might be good candidates for clinical trials evaluating adjuvant immune checkpoint inhibitors.
Citation Format: Griguolo G, Holgado E, Cortés J, Fasani R, Pascual T, Paré L, Bermejo B, Oliveira M, Morales S, Martinez N, Vidal M, Pernas S, Lopez R, Muñoz M, Galvan P, Garau I, Manso L, Alarcón J, Martínez E, Villagrasa P, LLombart-Cussac A, Prat A, Nuciforo P. Dynamics of tumor-infiltrating lymphocytes (TILs) during neoadjuvant dual HER2 blockade in HER2-positive (HER2+) breast cancer in the absence of chemotherapy [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P6-17-08.