THE EFFECTS OF CHANGES IN ENERGY BALANCE ON IMMUNE REGULATION AND TUMOR PROGRESSION IN THE 4T1.2 MAMMARY TUMOR MODEL
Open Access
- Author:
- Turbitt, William James
- Graduate Program:
- Integrative and Biomedical Physiology
- Degree:
- Doctor of Philosophy
- Document Type:
- Dissertation
- Date of Defense:
- June 15, 2017
- Committee Members:
- Connie Jo Rogers, Dissertation Advisor/Co-Advisor
Connie Jo Rogers, Committee Chair/Co-Chair
Matam Vijay Kumar, Committee Member
Kumble Sandeep Prabhu, Committee Member
Andrea Marie Mastro, Outside Member - Keywords:
- Cancer immunology
exercise
energy balance
4T1.2 model
metastasis
mammary tumor model - Abstract:
- One significant challenge in the field of breast cancer (BC) research is to determine how to reduce and/or eliminate the mortality associated with metastatic BC. Novel therapies, especially non-pharmacological, lifestyle-based interventions that prevent or slow metastatic disease with less severe side effects are greatly needed. Numerous lifestyle factors (including dietary components, body weight, and physical activity patterns) significantly impact BC risk and survival. Emerging population data suggests an inverse relation between physical activity and BC incidence, as well as an important role for exercise in the prevention of cancer recurrence and mortality. The observational nature of these studies limit the ability to determine biological mechanisms and the extent to which exercise, as opposed to changes in body weight, drive beneficial effects. Additionally, very little is known about the mechanisms contributing to the relation between physical activity and survival. Given the importance of metastases in the mortality of women with BC, understanding the role of exercise on metastatic burden may reveal important new targets for secondary and tertiary cancer prevention. The aim of study one was to control for weight and examine the effects of exercise, mild dietary restriction, or the combination of diet and exercise on the inflammation-immune axis and tumor progression in a preclinical metastatic BC model to determine the extent to which exercise or body weight contribute to cancer prevention. Dietary energy restriction-induced weight control (i.e., SED+ER mice) was effective at altering host splenic immunity and the expression of key genes in the tumor microenvironment (TME) related to immunosuppression and metastatic progression; however, this intervention failed to induce changes in primary tumor growth or spontaneous metastases. Moderate exercise in weight stable mice (EX+ER) resulted in a similar reduction in immunosuppressive and metastatic genes in the TME compared with the SED+ER mice; however, in addition, EX+ER mice had the greatest reduction in splenic immunosuppressive cells and plasma insulin-like growth factor-1 (IGF-1). The effects of moderate exercise in weight stable mice culminated in a significant delay in primary tumor growth and spontaneous metastases, suggesting that exercise-induced alterations in metabolic drivers of tumorigenesis, not simply a change in body weight, underlie the protective effects in the dual intervention group. Interestingly the exercise-induced protective effect on the emergence of immunosuppressive factors and reduced tumor burden was lost when mice continued to gain weight over the course of the study, suggesting that weight gain-induced disturbances on hormonal, inflammatory, and/or immunological function can override the exercise-induced benefits. Collectively, study one provided a deeper understanding of the extent to which exercise, and changes in body weight, underlies cancer protection. Few researchers have examined the effect of energy balance interventions on the efficacy of immunotherapeutic strategies. Two subsequent studies were designed to investigate the response to emerging cancer therapeutics in mice randomized to an energy balance paradigm (i.e., sedentary, ad libitum, weight gain [WG] group vs. exercising, mild dietary restriction, weight maintenance [WM] group) to identify potential mechanisms and provide translational support. Study two aimed to determine if there were any additive effects of moderate exercise in weight stable mice and the therapeutic administration of a broad-based, allogeneic, whole tumor cell cancer vaccine (VAX). There was a significant effect of both WM and VAX alone on primary tumor growth; and an additive effect of WM+VAX on primary tumor growth, lung and heart spontaneous metastases, splenocyte count at sacrifice, the number of total splenic myeloid-derived suppressor cells (MDSCs) and granulocytic subset of MDSCs, and plasma levels of IGF-1. Splenic interferon gamma (IFN) secretion in response to re-stimulation with tumor antigens was significantly elevated in response to VAX and WM; however, there was no additive effect of WM+VAX. These results suggested that our whole tumor cell cancer vaccine augmented the weight maintenance (via diet and exercise) effects on primary tumor growth and spontaneous metastasis; and suggested that vaccination may provide an immune stimulus to further promote the protective effects of moderate exercise alone in the metastatic 4T1.2 mammary tumor model. Study three aimed to determine if there were any additive effects of moderate exercise in weight stable mice and the dual therapeutic administration of a whole tumor cell cancer vaccine and programmed cell death protein-1 (PD-1) checkpoint blockade. We observed a cancer prevention effect of PD-1 checkpoint blockade in WG mice on primary tumor growth and spontaneous lung metastasis. However, moderate activity in weight stable mice, independent of PD-1 checkpoint blockade, was effective in reducing primary tumor growth and metastatic burden. The WM+PD-1 group displayed the lowest number of splenic MDSCs and granulocytic MDSCs and maintained its splenic lymphoid populations. Neither the number of tumor-infiltrating immune cells, the effector or activation status of tumor-infiltrating CD4+ helper and CD8+ cytotoxic T cells, nor functional outcomes were significantly different between groups. PD-1 checkpoint blockade in WG mice, moderate exercise in weight stable mice, and PD-1 checkpoint blockade in moderately exercising, weight stable mice showed comparable, albeit subtle differences, in tumor-immune crosstalk gene expression markers that drive the expansion of immunosuppressive cell types and impact metastatic progression. The lack of responsiveness to VAX+PD-1 checkpoint blockade in WM mice suggests that moderate exercise in weight stable mice may be enhancing antitumor immunity and/or reducing protumorigenic factors (i.e., similar mechanisms mediated by VAX+PD-1 checkpoint blockade). Results from the current studies provided insight into the extent to which exercise in weight stable mice underlie cancer protection. Also, results provided insight into potential mechanisms by which exercise can act via the inflammation-immune axis to attenuate the generation of a protumorigenic and immunosuppressive TME. These data demonstrated that preventing weight gain through diet and exercise may be an important recommendation to maintain prolonged antitumor effector responses and improve clinical outcomes. Results from the current studies provided insight into potential mechanisms by which physical activity exerts primary and secondary cancer prevention effects and provides a biological rationale for randomized clinical trials to investigate physical activity strategies to prevent metastatic progression in BC survivors and ultimately improve survival outcomes.