Abstract 520: Therapeutic antibodies against parathyroid hormone-related peptide (PTHrP) inhibit the growth of established skeletal metastasis in mice transplanted with human triple negative breast cancer cell lines

Abstract

Abstract Parathyroid Hormone-related Peptide (PTHrP)is a potent regulator of bone turnover and is thought to play a major role in the progression of skeletal metastasis through its activation of growth factors in the bone microenvironment. Our previous studies using genetically engineered mouse models in which PTHrP was ablated in the mammary epithelium have demonstrated that PTHrP also plays a direct role on tumor progression by enhancing tumor initiation, growth and metastasis outside the skeleton. We therefore proposed a novel mechanism of PTHrP driven bone metastasis by which PTHrP controls both the seed (tumor cells) and the soil (the bone microenvironment). We tested our hypothesis in triple negative breast cancer cell line (TNBC)models which are not responsive to either tamoxifen or Herceptin. Monoclonal antibodies against PTHrP were generated with strong in vitro anti-proliferative and anti-invasive potency in two human PTHrP expressing cell lines: the widely used TNBC MDAMB231 cell line and a patient derived TNBC cell line. Ablation of PTHrP or treatment with anti-PTHrP monoclonal antibodies induced major changes in the phenotype of these cells as determined by flow cytometry with stem cells and EMT markers, invasion assays and mammosphere assays indicating a reversal of their metastatic/invasive phenotype. Next we transplanted these cell lines into 8 weeks old female athymic nude mice to examine the efficacy of the therapeutic monoclonal antibodies on the progression of established bone metastasis.104 tumor cells were injected intra-tibially and animals examined at timed intervals(14, 21, 28, 25 and 42 days) with X-rays and high resolution CTs.At 14 days animals were administered intra-peritoneally with 200 micrograms of therapeutic anti-PTHrP antibodies or non-immune IgG(10 animals per group) and every 3 days thereafter til sacrifice(42 days). At sacrifice PET scan was performed to measure tumor volume and activity and bone were collected for histomorphometry analysis of tumor size and bone turnover activity. A significant reduction in bone lesions (P<0.05) was observed over time by imaging in animals treated with the therapeutic antibodies. At sacrifice PET scan indicates a strong reduction in tumor activity in treated animals (P<0.05). Histomorphometry analysis showed greater than 50% reduction in tumor volume associated with a significant inhibition of bone turnover in treated animals (P<0.05). In summary our data demonstrate that, in two TNBC models, PTHrP blockade can reprogram cancer cells to a reduced tumorigenicity and block the progression of established bone metastasis by targeting both the tumor cell and the bone microenvironment. Our data lend support to the “seed and soil” hypothesis proposed by Paget and the rational of anti-PTHrP therapeutic strategies alone or in combination with bone targeting agents in order to eradicate the development of skeletal metastasis. Citation Format: Jiarong Li, Anne Camirand, Mahvash Zakikhani, Karine Sellin, Richard Kremer. Therapeutic antibodies against parathyroid hormone-related peptide (PTHrP) inhibit the growth of established skeletal metastasis in mice transplanted with human triple negative breast cancer cell lines [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 520.