Abstract 2814: Systemic and local bone metastasis models for immuno-oncology drug development
Authors: Kähkönen Tiina E. , Suominen Mari I. , Mäki-Jouppila Jenni H. , Halleen Jussi M. , Scholz Arne , Bernoulli Jenni 
Published: August 2019

Abstract
Bone metastases are common and count 30-70% of metastases in breast, lung, and bladder cancer, and 80% of multiple myeloma patients have bone disease. Despite recent progress in cancer treatment, bone metastases remain incurable. Novel immunotherapies have the potential to cure also bone metastatic disease. Here we have established and validated syngeneic models with a focus on bone metastasis that could be used in preclinical efficacy studies. Syngeneic models were established for breast (4T1-GFP), multiple myeloma (5TGM1), bladder (MBT-2) and lung cancer (KLN-205). The cells were inoculated into systemic circulation (4T1 intracardially and 5TGM1 into tail vein) or bone marrow (MBT-2 and KLN-205). In the 4T1 model, tumor burden was assessed by ex vivo GFP imaging and in the 5TGM1 model by measuring serum IgG2b paraprotein levels during the study. Tumor-induced bone changes were followed by X-ray imaging in all models. Hind limbs were analyzed by histology and immunohistochemistry for tumor-infiltrating lymphocytes (TILs). The effects of standard-of-care compounds were assessed in the 4T1 (cyclophosphamide, 100 mg/kg or zoledronic acid, 0.1 mg/kg) and 5TGM1 (bortezomib, 1 mg/kg) models. The effect of anti-PD-1 treatment (200 µg/dose) was evaluated in the MBT-2 model. In the 4T1 model, osteolytic bone lesions formed within 13 days. In addition to bone metastases, about 50% of the mice had metastases in lungs, ovaries, kidneys and adrenal glands based on GFP imaging. Cyclophosphamide decreased the tumor burden and the area of osteolytic bone lesions. Zoledronic acid decreased the osteolytic lesion area but had no effect on tumor burden. Low number or no TILs were observed. In the 5TGM1 model, osteolytic lesions were observed and the study was ended at day 35. Metastases in ovaries, kidneys and adrenal glands were observed in about 30% of the mice. Bortezomib decreased serum paraprotein compared to vehicle treated mice. In both models, cachexia and paraplegia were occasionally observed. Moderate number of CD3+ TILs were observed in the tumors. In the intratibial MBT-2 and KLN-205 models, large osteolytic lesions were observed within 25 days, and in the KLN-205 model also lung metastases were observed. Anti-PD-1 treatment decreased osteolytic tumor area in the MBT-2 model. Moderate number of CD3+ and low number of CD4+ and CD8+ TILs were observed in the tumors growing in bone and also in the lung metastases. A high incidence of bone metastases was observed in all models. The use of systemic models allows studying the effects of test compounds in prevention or treatment of metastases. Intratibial models can be used when the primary interest is in tumor growth in bone microenvironment. Mimicking the clinical situation, none of the SOC compounds could prevent tumor growth completely, and therefore combination therapies are warranted for better overall efficacy.
Citation Format: Tiina E. Kähkönen, Mari I. Suominen, Jenni H. Mäki-Jouppila, Jussi M. Halleen, Arne Scholz, Jenni Bernoulli. Systemic and local bone metastasis models for immuno-oncology drug development [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2814.