Interrogating the Metabolic Interactions Between Adipocytes and Breast Cancer in a Patient-Derived Tissue Engineered Platform--Implications for the safety of autologous fat transfer in the setting of breast ductal carcinoma
Yoshiko Toyoda, BA, Karel-Bart Celie, BA, Justin Buro, BA, Alexandra Lin, BA, Jonathan Xu, BA, Andrew Miller, BA, John Morgan, PhD, Kristy A. Brown, PhD, Jason A. Spector, MD.
Weill Cornell Medical College, New York, NY, USA.
PURPOSE: Obesity is a known risk factor for the development of breast cancer (BC) and is understood to have a negative impact on prognosis. Obesity is associated with less response to BC therapy and more aggressive disease. Adipocytes have been identified as a source of exogenous lipids in many cancer cell types, and are thought to provide energy to fuel malignant survival and growth in BC. This relationship is of particular relevance in plastic surgery as the oncologic safety of autologous fat transfer, an increasingly ubiquitous adjunctive procedure for breast reconstruction after mastectomy, is largely unknown. Although clinical studies to examine this question are underway, an in vitro system is critical for elucidating the complex interplay between the cells that normally reside at the surgical recipient site. We have developed a 3D, patient-derived tissue engineered platform to directly assess the metabolic interactions among cells within the BC tumor microenvironment.
METHODS: Breast adipose tissue was acquired from patients undergoing breast reduction surgery. The tissue was enzymatically digested and sorted by differential centrifugation to retrieve adipocytes and ASCs. Polydimethylsiloxane wells were filled with type I 0.3% (w/v) collagen and seeded with varying concentrations of adipocytes labeled with the fluorescent lipid dye boron-dipyrromethene (BODIPY) and ASCs in the bulk, and fluorescently-labeled MDA-MB-231 BC cells on the surface. Cultures of BC cells in non-adipocyte containing collagen matrices served as controls. Lipid transfer and BC cell invasion into the collagen-adipocyte/ASC bulk matrices were analyzed using laser scanning confocal microscopy and image analysis.
RESULTS: As the BODIPY lipid stain was added to the adipocytes prior to seeding of the BC on the surface, any BODIPY staining seen within the BC cells must have originated from within the adipocytes. After 24 hrs of co-culture, the 3D collagen culture platform demonstrated BODIPY-stained mature adipocytes surrounded by stromal cells, akin to the native architecture in human breast tissue. At the interface of the cancer cells with the stroma, lipid transfer was observed from adipocytes to BC cells as demonstrated by the change in the morphology of BC cells in proximity to the lipid-filled adipocytes from a spindle-like shape to more round appearance, filled centrally with green fluorescent lipid droplets and the cytosol pushed to the periphery.
CONCLUSION: We have established a novel 3D platform to study BC microenvironment, including metabolic interactions between primary human breast adipocytes and BC cells. Transfer of fluorescently-labeled lipids directly from adipocytes to BC cells may induce aberrant metabolism to fuel malignant growth and adaptive survival, while the presence or absence of ASCs and adipocytes enables analysis of their effect on metastatic progression. Our novel, 3D platform can untangle the complex interplay within the entire breast cancer tumor microenvironment for high-throughput analysis and can help elucidate the safety of adipose tissue transfer, with and without ASC enrichment, in breast reconstruction in post-oncologic breast reconstruction.
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