Abstract P6-17-07: Gene signatures and subtype changes during HER2 dual blockade in PAM50 HER2-enriched HER2-positive breast cancer
Authors: Brasó-Maristany F , Griguolo G , Llombart-Cussac A , 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 , Cortés J , Prat A 
Published: May 2019

Abstract
Background: HER2-positive (HER2+) breast cancer (BC) is composed of 4 molecular subtypes: Luminal A and B, HER2-enriched (HER2-E) and Basal-like. Among them, the HER2-E is highly sensitive to anti-HER2 treatment. However, ˜60% of HER2-E tumors do not achieve a pathological complete response (pCR) following neoadjuvant dual HER2 blockade without chemotherapy. Here, we aimed to better understand the molecular changes of the HER2-E subtype during anti-HER2 treatment.
Methods: Gene expression was evaluated in 101 patients with HER2-E tumors from the PAMELA neoadjuvant phase II trial (Lancet Oncol 2017). Briefly, women with HER2+ BC were treated with lapatinib and trastuzumab (and hormonal therapy if hormone receptor [HR]-positive) for 18 weeks. The median time between the last dose of treatment and surgery was 35 days (range=213; interquartile range=16). Expression of the PAM50 genes and 6 PAM50 signatures (Luminal A, Luminal B, HER2-E, Basal-like, normal-like and the PAM50 proliferation score) were determined using the nCounter platform at baseline (n=101), after 2 weeks of treatment (n=96) and in residual tumors (non-pCR) at surgery (n=57). Same analyses were done in 2 HER2+/HER2-E cell line models (BT474 [HR+] and SKBR3 [HR-]) following in vitro treatment with trastuzumab in combination with lapatinib. Biological changes between 2 time-points were determined by paired t-tests with a false discovery rate (FDR) &lt;5%.
Results: After 2 weeks of treatment, 85.7% and 94.6% of the 56 genes/signatures were found differentially expressed (FDR&lt;5%) in HER2-E/HR+ (n=35) and HER2-E/HR- (n=61) tumors, respectively. The two gene lists were highly correlated (correlation coefficient=0.93). Overall, a significant relative increase in Luminal A and normal-like signature scores, and a relative decrease in proliferation, HER2-E and Luminal B signature scores, were observed between baseline and week 2. Interestingly, a PAM50 subtype switch to Luminal A was observed in 31.6% and 4.8% of HER2-E/HR+ and HER2-E/HR- tumors. In BT474 and SKBR3, all genes/signatures were also found differentially expressed (FDR&lt;5%) following 72h of dual HER2 blockade. The in vitro findings recapitulated the in vivo findings in 80-86% of the genes/signatures. Similar to tumors, a switch to a Luminal A subtype following dual HER2 blockade was observed in BT474 but not in SKBR3. Finally, 92.9% of the 56 genes/signatures were found differentially expressed (FDR&lt;5%) in residual tumors at surgery compared to week 2. Contrary to the findings in the first 2 weeks of treatment, a general rebound effect in gene expression was observed between week 2 and surgery. Similarly, a rebound effect was observed in 60% of the genes/signatures in BT474 after removing anti-HER2 therapy for 72h, leading to a subtype switch from Luminal A back to HER2-E.
Conclusions: Dual HER2 blockade in the HER2-E subtype induces large biological changes that lead to a more low-proliferative Luminal A phenotype both in tumors and in vitro models, especially in HER2-E/HR+ disease. These phenotypic changes are reversible upon stopping anti-HER2 treatment. This finding supports the use of maintenance anti-HER2 treatment +/- endocrine therapy (if HR+) in advanced HER2+ BC.
Citation Format: Brasó-Maristany F, Griguolo G, Llombart-Cussac A, 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, Cortés J, Prat A. Gene signatures and subtype changes during HER2 dual blockade in PAM50 HER2-enriched HER2-positive breast cancer [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 P6-17-07.