title: Mimicking the tumor microenvironment to regulate macrophage phenotype and assessing chemotherapeutic efficacy in embedded cancer cell/macrophage spheroid models.
abstract: Tumor associated macrophages (TAMs) are critical stromal components intimately involved with the progression, invasion, and metastasis of cancer cells. To address the need for an in vitro system that mimics the clinical observations of TAM localizations and subsequent functional performance, a cancer cell/macrophage spheroid model is described. The central component of the model is a triple negative breast cancer spheroid embedded in a three-dimensional collagen gel. Macrophages are incorporated in two different ways. The first is a heterospheroid, a spheroid containing both tumor cells and macrophages. The heterospheroid mimics the population of TAMs infiltrated into the tumor mass, thus being exposed to hypoxia and metabolic gradients. In the second model, macrophages are diffusely seeded in the collagen surrounding the spheroid, thus modeling TAMs in the cancer stroma. The inclusion of macrophages as a heterospheroid changes the metabolic profile, indicative of synergistic growth. In contrast, macrophages diffusely seeded in the collagen bear the same profile regardless of the presence of a tumor cell spheroid. The macrophages in the heterospheroid secrete EGF, a cytokine critical to tumor/macrophage co-migration, and an EGF inhibitor decreases the metabolic activity of the heterospheroid, which is not observed in the other systems. The increased secretion of IL-10 indicates that the heterospheroid macrophages follow an M2/TAM differentiation pathway. Lastly, the heterospheroid exhibits resistance to paclitaxel. In summary, the collagen embedded heterospheroid model promotes TAM-like characteristics, and will be of utility in cancer biology and drug discovery.
  
  
  
  Two in vitro collagen-embedded multicellular spheroid models are described that mimic the clinical observations of macrophage localization within a tumor. Incorporation of macrophages within a breast cancer spheroid emphasizes cell-cell interactions with subsequent differentiation toward a tumor-promoting TAM phenotype. In contrast, macrophages seeded around the tumor spheroid display decreased interaction with cancer cells and no indication of a TAM phenotype. Finally, the presence of macrophages in the heterospheroid increases resistance to paclitaxel. This study demonstrates that cell-cell interactions and 3D collagen matrix direct macrophage activity, and, thus, highlights the important role the local environment itself plays in macrophage behavior.
authors.affiliation: Department of Biomedical Engineering, Cummington Street, Boston University, Boston, MA 02215, United States.
authors.forename: Kristie M
authors.initials: KM
authors.lastname: Tevis
authors.affiliation: Department of Biomedical Engineering, Cummington Street, Boston University, Boston, MA 02215, United States.
authors.forename: Ryan J
authors.initials: RJ
authors.lastname: Cecchi
authors.affiliation: Division of Thoracic Surgery, Department of Surgery, Brigham and Women's Hospital, Boston, MA 02215, United States. Electronic address: Yolonda_Colson@dfci.harvard.edu.
authors.forename: Yolonda L
authors.initials: YL
authors.lastname: Colson
authors.affiliation: Department of Biomedical Engineering, Cummington Street, Boston University, Boston, MA 02215, United States; Department of Chemistry, Metcalf Center for Science and Engineering, Boston University, Boston, MA 02215, United States; Department of Medicine, 715 Albany Street, Boston University, Boston, MA 02118, United States. Electronic address: mgrin@bu.edu.
authors.forename: Mark W
authors.initials: MW
authors.lastname: Grinstaff
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002450:Cell Communication / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D045744:Cell Line, Tumor; D003094:Collagen / Q000494:pharmacology; D016207:Cytokines / Q000378:metabolism; D019008:Drug Resistance, Neoplasm / Q000187:drug effects; D005260:Female; D006801:Humans; D008264:Macrophages / Q000473:pathology*; D051379:Mice; D008954:Models, Biological; D010641:Phenotype; D000067996:RAW 264.7 Cells; D018874:Spheroids, Cellular / Q000187:drug effects / Q000473:pathology*; D059016:Tumor Microenvironment / Q000187:drug effects
chemical_list: D000970:Antineoplastic Agents; D016207:Cytokines; D003094:Collagen
keywords: Breast cancer; Heterospheroid; Paclitaxel; Spheroid; Tumor associated macrophages