Abstract 2036: Osteoblasts use multiple mechanisms of cellular communication to regulate breast cancer cell proliferation in bone
Authors: Kolb Alexus D. , Shupp Alison B. , Mukhopdhyay Dimpi , Bussard Karen M. 
Published: August 2019

Abstract
Breast cancer (BC) preferentially metastasizes to bone. While the mechanism for preferred metastasis is unknown, bone likely provides a hospitable environment that attracts BC cells (BCCs) and allows them to colonize and grow. Bone metastases perturb the balance of normal bone remodeling. Emerging evidence suggests that OBs may be altered and respond differently to cancer cells during metastatic BC progression. According to one study, OBs suppress cancer cell growth in early metastasis. Our work suggests OBs reduce BCC growth in bone prior to macrometastatic colonization. Thus, our data suggest OBs have tumor-inhibitory properties. We have new evidence to suggest that communication between OBs and BCCs ‘educates’ OBs to produce factors that suppress BCC proliferation in bone. We have in-vitro and in-vivo mouse-model evidence that ‘educated’ osteoblasts (EOs) have a unique secretory protein profile compared to ‘uneducated’ OBs. We also identified EOs as being present in the bone tissue samples of human patients with bone metastatic BC via multi-plex immunofluorescence. When we treated BCCs with EO conditioned media (CM), BCC proliferation was reduced in both triple negative and ER+ metastatic BCCs, while CM from ‘uneducated’ OBs did not affect BCC proliferation. This effect was mediated through alterations in EO production of decorin and NOV. We identified EO CM as a rich source of exosomes (exo) and confirmed the presence of an exo population via iodixanol density gradient and western blotting for specific exo protein markers. We found that EO-derived exo, but not ‘uneducated’ OB-derived exo, decreased proliferation of ER+ and triple negative BCC. Also, exposure to EO-derived exo 1) decreased the number of Ki67 positive metastatic BCC, and 2) decreased the number of metastatic BCC in the S phase of cell cycle as measured by EdU incorporation. Moreover, we labeled EO exo with RFP-conjugated CD63 to visually confirm exo transfer from EO cells to BCCs using confocal microscopy. And, co-culture with EOs increased triple negative and ER+ metastatic BC expression of p21 compared to co-cultures with ‘uneducated’ OBs. Our data suggest that EOs use multiple mechanisms of cellular communication to regulate BCC proliferation in bone. Impact: Our data suggest OBs produce factors that suppress metastatic BCC growth. Much less attention has been given to OB interactions with tumor cells at sites of bone metastasis due to observations that OB populations are reduced at sites of advanced osteolysis. However, we propose that OBs may be valuable endogenous targets to aid in suppression of metastatic BC growth in the niche in concert with therapeutic drugs to kill the cancer cells. Our data suggest there is a population of OBs that demonstrate a functional role in suppressing metastatic BCC growth; a property capable of exploitation. For these reasons, OBs and EOs are strong candidates for therapeutic targeting.
Citation Format: Alexus D. Kolb, Alison B. Shupp, Dimpi Mukhopdhyay, Karen M. Bussard. Osteoblasts use multiple mechanisms of cellular communication to regulate breast cancer cell proliferation in bone [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 2036.