Data from: Effect of pancreatic cancer-derived extracellular vesicles on bone marrow-derived macrophages

Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive
cancers, with a majority of patients presenting with metastatic disease at
diagnosis. Extracellular vesicles (EVs) released by cancer cells have
emerged as key mediators of intracellular
signaling, communication, and immune modulation within the tumor
microenvironment (TME).  Tumor-associated macrophages
(TAMs) are crucial components of TME that influence tumor growth and
metastasis. The present study is focused on determining
the role of PDAC-derived EVs in altering macrophage phenotype, function,
and metabolism, and their impact on tumor cell proliferation and invasion.
EVs were isolated from the conditioned medium that was used to culture one
established pancreatic cancer cell (PANC-1), one PDX cell line (PPCL68),
and a normal (non-tumorigenic) established pancreatic epithelial cell line
(hTERT-HPNE), using size exclusion chromatography. The EVs were
extensively characterized using nanoparticle tracking (NTA),
cryo-electron microscopy, and immunoblot analysis as per
ISEV guidelines. Macrophages were generated from the bone marrow cells of
C57/BL6 mice cultured in vitro using macrophage colony-stimulating factor
(M-CSF). Macrophages were co-cultured with isolated PDAC or normal
cell-derived EVs. Flow cytometry and multi-omic analyses were performed to
determine the effect of EVs on macrophage phenotype, function, and
behavior. Our results show that PDACcell-derived EVs were taken up by the
macrophage in a time-dependent manner that resulted in an
immunosuppressive phenotype characterized by higher expression of CD206
and PD-L1 and higher secretion of immunosuppressive cytokines, including
TGF-beta, IL-10, and GM-CSF.  Functionally, PDAC-EV-treated
macrophages were able to suppress the proliferation of CD8 T cells under
in vitro and in vivo conditions. Metabolically, cancer EV-treated
macrophages showed accumulation of immunosuppressive metabolites.