Breast cancer preferentially metastasizes to the bone, where the five-year relative survival rate falls from 90% to <10%. Although the precise mechanism underlying preferential metastasis is unknown, the bone likely provides a hospitable environment that both attracts breast cancer cells and allows them to colonize and grow. Clinical evidence suggests that breast cancer cells can exist in a dormant state in the bone for up to 25 years, evading immune surveillance and chemotherapeutic treatments which target cycling cells. As a result, breast cancer patients successfully treated for primary breast cancer decades before develop overt macrometastases without prior symptoms. Thus, there is a critical need for elucidating molecular mechanisms facilitating breast cancer dormancy and reactivation. This proposal aims to understand the role of the bone microenvironment in breast cancer cell bone engraftment (Aim 1), dormancy (Aim 2), and reactivation (Aim 3), with the long-term goal of identifying key interactions for therapeutic intervention between the bone microenvironment, bone stromal cells, and breast cancer cells that contribute to breast cancer cell bone dormancy and metastasis. To address these challenges, first, in Aim 1 (mentored), I will determine the importance of bone stromal cells and their secreted factors for metastatic breast cancer cell bone engraftment and colonization. In this aim, I will utilize in-vitro cell culure and novel compartmentalized bioreactor and in-vivo extramedullary bone models that mimic human breast cancer metastasis in-vivo to determine if 1) breast cancer cells form gap junctions with bone stromal cells, 2) alterations in mesenchymal stem cell-derived factors regulate breast cancer cell engraftment, or 3) if formation of cancer-associated fibroblasts is modulated by osteoblast-derived cytokines. In the Second Aim (Independent), I will identify key interactions between bone stromal cells and breast cancer cells that lead to breast cancer cell dormancy in the bone. I will 1) use bone-seeking breast cancer cells and the spontaneous mouse model of breast cancer bone metastasis to identify gap junctions critical to breast cancer cell quiescence. Next, I will 2) determine how crosstalk between osteoblasts and cancer-associated fibroblasts ultimately affects their functions. Finally, I will 3) use cell culture to determine if cancer-associated fibroblasts and osteoblasts express exosomes that impact breast cancer cell dormancy. Lastly, in the Third Aim (Independent), I will uncover mechanisms involving bone stromal cells that contribute towards the reactivation of breast cancer cells and formation of a metastatic niche in bone. Here, I will use 1) short-hairpin RNAs to identify specific osteoblast-derived cytokines that reactivate breast cancer cells utilizing both co-culture and an intratibial mouse model. Additionally, I will use the compartmentalized bioreactor to determine if cancer-associated fibroblasts regulate 2) the differentiation or 3) mediate apoptosis of osteoblasts. Finally, I will 4) determine the importance of cancer-associated fibroblasts in the formation of osteoclasts.
Breast cancer recurrence in the bone after therapy and long periods of remission is frequent, and the exact molecular mechanisms for breast cancer cell dormancy and reactivation are unknown. This proposal underscores the importance of investigating the bone microenvironment as a critical regulator in breast cancer cell dormancy. Results generated from this proposal are ultimately aimed at identifying therapeutic targets to eradicate dormant breast cancer cells, allowing for an improved quality of life and longer time of survival for individuals with breast cancer. The written critiques of individual reviewers are provided in essentially unedited form in this section. Please note that these critiques and criteria scores were prepared prior to the meeting and may not have been revised subsequent to any discussions at the review meeting. The Resume and Summary of Discussion section above summarizes the final opinions of the committee.
| Bussard, Karen M; Mutkus, Lysette; Stumpf, Kristina et al. (2016) Tumor-associated stromal cells as key contributors to the tumor microenvironment. Breast Cancer Res 18:84 |
| Coleman, Kate E; Grant, Gavin D; Haggerty, Rachel A et al. (2015) Sequential replication-coupled destruction at G1/S ensures genome stability. Genes Dev 29:1734-46 |
| Rizzardi, Lindsay F; Coleman, Kate E; Varma, Dileep et al. (2015) CDK1-dependent inhibition of the E3 ubiquitin ligase CRL4CDT2 ensures robust transition from S Phase to Mitosis. J Biol Chem 290:556-67 |
