Abstract P2-05-21: Predictive value of quantitative HER2 and HER3 levels combined with downstream signaling markers in HER2-positive advanced breast cancer patients treated with lapatinib
Authors: Duchnowska R , Sperinde J , Czartoryska-Arlukowicz B , Mysliwiec P , Winslow J , Radecka B , Petropoulos C , Demlova R , Orlikowska M , Kowalczyk A , Lang I , Ziólkowska B , Debska-Szmich S , Merdalska M , Grela-Wojewoda A , Zawrocki A , Biernat W , Huang W , Jassem J 
Published: March 2017

Abstract
Background: Clinical correlates of lapatinib resistance have not been well defined. Previous studies implicated genes regulated by the estrogen receptor (ER) and activation or mutation of proteins downstream from HER family receptors. In the current study, HER2 and HER3 expression levels were quantitatively measured using a VeraTag® fluorescence-based assay, in addition to seven downstream signaling proteins determined by IHC. All biomarkers were correlated with overall survival (OS) in patients treated with lapatinib.
Methods: Formalin-fixed, paraffin-embedded samples were obtained from the primary tumor of 191 patients treated with lapatinib plus capecitabine following progression on trastuzumab. The HERmark® Breast Cancer Assay (Monogram Biosciences, South San Francisco) was used to quantify HER2 protein expression levels. HER3 protein expression was quantified using the VeraTag® technology (Monogram Biosciences). Expression of ER, PTEN, Cyclin E, HIF-2alpha, p-p70S6K, p-AMPK and p-MAPK were determined by IHC (Duchnowska et al., Oncotarget 2016; 7:550). OS analyses of HER2 and HER3 were stratified by key clinical variables, including stage and presence of a brain metastasis prior to lapatinib-based therapy.
Results: Among the downstream signaling molecules, HIF-2alpha (r = -0.23; p = 0.047) and ER (r = -0.27; p = 0.005) were negatively correlated with HER2 expression after adjustment for multiple testing. PTEN appeared to correlate with HER3, but was not significant after adjustment for multiple testing. OS was significantly shorter for both those below the cut-off level of positivity by the HERmark assay (HR = 1.8; p = 0.029), and those with above median HER2 levels (HR = 1.7; p = 0.009), as compared to cases with in between levels. The relationship between HER2 and OS is also captured by a U-shaped, parabolic function in HER2 (p = 0.005). Elevated HER3 showed a trend toward a correlation with longer OS (HR = 0.66/log; p = 0.16), somewhat stronger in the ER-negative subset (HR = 0.55/log; p = 0.085) and in the subset with above-median HER2 (0.48/log; p = 0.10), where inhibiting HER2 activation of HER3 may be more important. In multivariate Cox models, HER2 (parabola, intermediate HER2 best, p = 0.001), presence of brain metastases (HR = 2; p &lt; 0.001), ER (HR = 0.60; p = 0.009) and either p-p70S6K (HR = 0.66; p = 0.018) or p-AMPK (HR = 0.67; p = 0.022) were significantly associated with OS (p-p70S6K and p-AMPK were mutually correlated).
Conclusions: Patients with moderately increased HER2 levels may have best outcomes while receiving lapatinib following progression on trastuzumab. This supports recent findings of a less benefit from lapatinib in patients with high HER2 expression (Nunciforo et al., SABCS 2015, P3-07-08). HER3 levels do not seem to substantially impact the prognosis. Further studies are warranted to explore the predictive utility of quantitative HER2 and HER3 in guiding HER2-directed therapies.
Citation Format: Duchnowska R, Sperinde J, Czartoryska-Arlukowicz B, Mysliwiec P, Winslow J, Radecka B, Petropoulos C, Demlova R, Orlikowska M, Kowalczyk A, Lang I, Ziólkowska B, Debska-Szmich S, Merdalska M, Grela-Wojewoda A, Zawrocki A, Biernat W, Huang W, Jassem J. Predictive value of quantitative HER2 and HER3 levels combined with downstream signaling markers in HER2-positive advanced breast cancer patients treated with lapatinib [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P2-05-21.