title: Tumor Endothelial Cells with Distinct Patterns of TGFβ-Driven Endothelial-to-Mesenchymal Transition.
abstract: Endothelial-to-mesenchymal transition (EndMT) occurs during development and underlies the pathophysiology of multiple diseases. In tumors, unscheduled EndMT generates cancer-associated myofibroblasts that fuel inflammation and fibrosis, and may contribute to vascular dysfunction that promotes tumor progression. We report that freshly isolated subpopulations of tumor-specific endothelial cells (TEC) from a spontaneous mammary tumor model undergo distinct forms of EndMT in response to TGFβ stimulation. Although some TECs strikingly upregulate α smooth muscle actin (SMA), a principal marker of EndMT and activated myofibroblasts, counterpart normal mammary gland endothelial cells (NEC) showed little change in SMA expression after TGFβ treatment. Compared with NECs, SMA(+) TECs were 40% less motile in wound-healing assays and formed more stable vascular-like networks in vitro when challenged with TGFβ. Lineage tracing using ZsGreen(Cdh5-Cre) reporter mice confirmed that only a fraction of vessels in breast tumors contain SMA(+) TECs, suggesting that not all endothelial cells (EC) respond identically to TGFβ in vivo. Indeed, examination of 84 TGFβ-regulated target genes revealed entirely different genetic signatures in TGFβ-stimulated NEC and TEC cultures. Finally, we found that basic FGF (bFGF) exerts potent inhibitory effects on many TGFβ-regulated genes but operates in tandem with TGFβ to upregulate others. ECs challenged with TGFβ secrete bFGF, which blocks SMA expression in secondary cultures, suggesting a cell-autonomous or lateral-inhibitory mechanism for impeding mesenchymal differentiation. Together, our results suggest that TGFβ-driven EndMT produces a spectrum of EC phenotypes with different functions that could underlie the plasticity and heterogeneity of the tumor vasculature.
authors: Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.
authors: Lin
authors: L
authors: Xiao
authors: Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.
authors: Dae Joong
authors: DJ
authors: Kim
authors: Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.
authors: Clayton L
authors: CL
authors: Davis
authors: Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.
authors: James V
authors: JV
authors: McCann
authors: Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.
authors: James M
authors: JM
authors: Dunleavey
authors: Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.
authors: Alissa K
authors: AK
authors: Vanderlinden
authors: Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.
authors: Nuo
authors: N
authors: Xu
authors: Center for Integrative Chemical Biology and Drug Discovery, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.
authors: Samantha G
authors: SG
authors: Pattenden
authors: Center for Integrative Chemical Biology and Drug Discovery, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.
authors: Stephen V
authors: SV
authors: Frye
authors: Department of Surgery, Duke University, Durham, North Carolina.
authors: Xia
authors: X
authors: Xu
authors: Department of Surgery, Duke University, Durham, North Carolina.
authors: Mark
authors: M
authors: Onaitis
authors: Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.
authors: Elizabeth
authors: E
authors: Monaghan-Benson
authors: Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina. Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina. McAllister Heart Institute, Chapel Hill, North Carolina.
authors: Keith
authors: K
authors: Burridge
authors: Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina. Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina. McAllister Heart Institute, Chapel Hill, North Carolina. acdudley@med.unc.edu.
authors: Andrew C
authors: AC
authors: Dudley
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002465:Cell Movement; D002469:Cell Separation; D054338:Cell Transdifferentiation; D042783:Endothelial Cells / Q000502:physiology*; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000502:physiology; D008325:Mammary Neoplasms, Experimental / Q000473:pathology; D008822:Mice, Transgenic; D009368:Neoplasm Transplantation; D016212:Transforming Growth Factor beta / Q000502:physiology*
chemical_list: D016212:Transforming Growth Factor beta; D016222:Fibroblast Growth Factor 2
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