Abstract P5-05-06: Development of advanced pre-clinical in vivo models of metastatic breast cancer
Authors: Okano M , Kawaguchi T , Okano I , Katsuta E , Takabe K 
Published: February 2018

Abstract
Backgrounds: The fact that we continue to lose 40,000 women with breast cancer every year in the US despite the recent advance in basic research clearly demonstrate disconnect in translation of basic research findings to clinic. This is largely due to lack of appropriate animal model that mimic clinical conditions for preclinical studies that result in high failure rate of clinical trials. To date, we had established many syngeneic mouse models, which are not free from limitations; 1) few clinically relevant animal models with bone metastasis have been established, 2) syngeneic mouse model cannot address human cancer genomics and tumor heterogeneity. Patient-Derived Xenograft (PDX) model has emerged as pre-clinical model to address these issues, however, it suffers low tumor take rate of around 20-40%, and lack metastatic model. Here, we describe development of orthotopic implantation, and bone and liver metastatic breast cancer mouse models to overcome these limitations.
Methods: 1) 4T1.2-luc3 cells that has metastatic potential to the bone were orthotropically inoculated as a syngeneic mouse model, imaged with IVIS and MRI. 2) Patient tumor tissues of 1mm(3) were implanted surgically into dorsal subcutaneous space (SQ), or orthotropically into mammary fat pat #2 and #4 (MFP).
Results: 1) We established a syngeneic breast cancer bone metastasis model. Primary tumors were surgically resected days after 4T1.2-luc3 cells were orthotopically implanted under direct vision. Removal of primary tumor allowed bioluminescent visualization and quantification of bone metastasis by IVIS. We found that MRI was effective in evaluating bone metastasis and bone related events in these mice. MRI allows differentiation of bone metastasis from metastasis to the surrounding organs with bone destruction image, whereas conventional bioluminescence imaging shows only existence of cancer cells. 2) The overall tumor take rate of the tumor in PDX model was 46.0% (74/161 implantation site). Take rate from triple-negative breast cancer tumors was 56.1% (74/132), on the other hand, that from ER positive tumors was 0% (0/39). Tumor take rate was significantly better in MFP implantation than SQ (39.5%, 30/76 vs 51.2%, 44/85, p&lt;0.01). Tumor weight were significantly heavier in MFP compared to SQ (0.072g vs 0.328g, p&lt;0.00001). With more passage, the difference in tumor weight between SQ and MFP was significantly increased(p&lt;0.0001). Finally, we developed a PDX breast cancer liver metastasis model by surgically implanting tissue fragment into liver using direct vision technique. We found MRI to be very useful as a living imaging modality to evaluate cancer progression in the deeply located metastatic sites of PDX models.
Conclusions: We have established orthotropic syngeneic breast cancer bone metastasis model as well as improved breast cancer PDX model with synchronous liver metastasis utilizing MRI. Our novel models could be powerful tools for preclinical studies.
Citation Format: Okano M, Kawaguchi T, Okano I, Katsuta E, Takabe K. Development of advanced pre-clinical in vivo models of metastatic breast cancer [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P5-05-06.