title: Pancreatic cancer-associated stellate cells promote differentiation of myeloid-derived suppressor cells in a STAT3-dependent manner.
abstract: Pancreatic stellate cells (PSC) are a subset of pancreatic cancer-associated fibroblasts. These cells provide prosurvival signals to tumors; however, little is known regarding their interactions with immune cells within the tumor microenvironment. We hypothesized that factors produced by human PSC could enhance myeloid-derived suppressor cell (MDSC) differentiation and function, which promotes an immunosuppressive microenvironment. Primary PSC cell lines (n = 7) were generated from human specimens and phenotypically confirmed via expression of vimentin, α-smooth muscle actin (α-SMA), and glial fibrillary acidic protein (GFAP). Luminex analysis indicated that PSC but not human fetal primary pancreatic fibroblast cells (HPF; negative controls) produced MDSC-promoting cytokines [interleukin (IL-6), VEGF, macrophage colony-stimulating factor (M-CSF) ] and chemokines (SDF-1, MCP-1). Culture of peripheral blood mononuclear cells [peripheral blood mononuclear cell (PBMC), n = 3 donors] with PSC supernatants or IL-6/granulocyte macrophage colony-stimulating factor (GM-CSF; positive control) for 7 days promoted PBMC differentiation into an MDSC (CD11b+CD33+) phenotype and a subpopulation of polymorphonuclear CD11b+CD33+CD15+ cells. The resulting CD11b+CD33+ cells functionally suppressed autologous T-lymphocyte proliferation. In contrast, supernatants from HPF did not induce an MDSC phenotype in PBMCs. Culture of normal PBMCs with PSC supernatants led to STAT3 but not STAT1 or STAT5 phosphorylation. IL-6 was an important mediator as its neutralization inhibited PSC supernatant-mediated STAT3 phosphorylation and MDSC differentiation. Finally, the FLLL32 STAT3 inhibitor abrogated PSC supernatant-mediated MDSC differentiation, PSC viability, and reduced autocrine IL-6 production indicating these processes are STAT3 dependent. These results identify a novel role for PSC in driving immune escape in pancreatic cancer and extend the evidence that STAT3 acts as a driver of stromal immunosuppression to enhance its interest as a therapeutic target.
authors.affiliation: Division of Medical Oncology, Department of Internal Medicine, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Mace
authors.forename: Zeenath
authors.initials: Z
authors.lastname: Ameen
authors.forename: Amy
authors.initials: A
authors.lastname: Collins
authors.forename: Sylwia
authors.initials: S
authors.lastname: Wojcik
authors.forename: Markus
authors.initials: M
authors.lastname: Mair
authors.forename: Gregory S
authors.initials: GS
authors.lastname: Young
authors.forename: James R
authors.initials: JR
authors.lastname: Fuchs
authors.forename: Tim D
authors.initials: TD
authors.lastname: Eubank
authors.forename: Wendy L
authors.initials: WL
authors.lastname: Frankel
authors.forename: Tanios
authors.initials: T
authors.lastname: Bekaii-Saab
authors.forename: Mark
authors.initials: M
authors.lastname: Bloomston
authors.forename: Gregory B
authors.initials: GB
authors.lastname: Lesinski
mesh_terms: D002454:Cell Differentiation; D045744:Cell Line, Tumor; D006801:Humans; D015850:Interleukin-6 / Q000502:physiology; D008213:Lymphocyte Activation; D022423:Myeloid Cells / Q000166:cytology / Q000276:immunology*; D010190:Pancreatic Neoplasms / Q000276:immunology* / Q000473:pathology; D058954:Pancreatic Stellate Cells / Q000473:pathology / Q000502:physiology*; D050796:STAT3 Transcription Factor / Q000502:physiology*; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000276:immunology; D019139:Tumor Escape
chemical_list: D015850:Interleukin-6; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human