Prospective neoadjuvant analysis of PET imaging and mechanisms of resistance to Trastuzumab shows role of HIF1 and autophagy

Background: Although Trastuzumab has improved survival of HER2+ breast cancer patients, resistance to the agent pre-exists or develops through the course of therapy. Here we show that a specific metabolism and autophagy-related cancer cell phenotype relates to resistance of HER2+ breast cancer to Trastuzumab and chemotherapy. Methods: Twenty-eight patients with locally advanced primary breast cancer were prospectively scheduled to received one cycle of Trastuzumab followed by a new biopsy on day 21, followed by taxol/Trastuzumab chemotherapy for four cycles before surgery. FDG PET/CT scan was used to monitor tumour response. Tissue samples were immunohistochemically analysed for metabolism and autophagy markers. Results: In pre-Trastuzumab biopsies, the LC3A+/HER2+ cell population was correlated with HIF1 alpha expression (P=0.01), while GLUT1 and LC3B expression were correlated with Ki67 proliferation index (P=0.01 and P=0.01, respectively). FDG PET tumour dimensions before therapy were correlated with LC3B expression (P=0.005). Administration of Trastuzumab significantly reduced clinical and PET-detected tumour dimensions (P < 0.01). An inverse association of tumour response with the percentage of cells expressing HIF1a at baseline was documented (P=0.01). Administration of Trastuzumab resulted in a decrease of the proliferation index (P=0.004), GLUT1 (P=0.04) and HER2 (P=0.01) expression. In contrast, the percentage of LC3A + /HER2 + cells was increased (P=0.01). High baseline HIF1a expression was the only parameter associated with poorer pathological response to preoperative chemotherapy (P=0.001). Conclusions: As the HER2 + /LC3A + phenotype, which often overexpresses HIF1 alpha, is a major subpopulation increasing after therapy with Trastuzumab, LC3A- and HIF1 alpha-targeting therapies should be investigated for the augmentation of anti-HER2 therapy efficacy.