Tumor Endothelial Cells with Distinct Patterns of TGFβ-Driven Endothelial-to-Mesenchymal Transition

Xiao, Lin; Kim, Dae Joong; Davis, Clayton L.; McCann, James V.; Dunleavey, James M.; Vanderlinden, Alissa K.; Xu, Nuo; Pattenden, Samantha G.; Frye, Stephen V.; Xu, Xia; Onaitis, Mark W.; Monaghan-Benson, Elizabeth; Burridge, Keith; Dudley, Andrew C.

doi:10.1158/0008-5472.can-14-1616

Cited by 63 publications

(67 citation statements)

References 49 publications

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“…3A), with comparable effects at 1 and 10 ng ml −1 TGFβ2. All following experiments were carried out with 10 ng ml −1 TGFβ2, in line with previous in vitro studies on EndMT 31, 32 . Ligand specificity of BMP9 (1 ng ml −1 ) and TGFβ2 (10 ng ml −1 ) was shown by selective induction of their target genes ENG and CNN1 , respectively (Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

Dynamic regulation of canonical TGFβ signalling by endothelial transcription factor ERG protects from liver fibrogenesis

et al. 2017

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The role of the endothelium in protecting from chronic liver disease and TGFβ-mediated fibrosis remains unclear. Here we describe how the endothelial transcription factor ETS-related gene (ERG) promotes liver homoeostasis by controlling canonical TGFβ-SMAD signalling, driving the SMAD1 pathway while repressing SMAD3 activity. Molecular analysis shows that ERG binds to SMAD3, restricting its access to DNA. Ablation of ERG expression results in endothelial-to-mesenchymal transition (EndMT) and spontaneous liver fibrogenesis in EC-specific constitutive hemi-deficient (Erg cEC-Het) and inducible homozygous deficient mice (Erg iEC-KO), in a SMAD3-dependent manner. Acute administration of the TNF-α inhibitor etanercept inhibits carbon tetrachloride (CCL4)-induced fibrogenesis in an ERG-dependent manner in mice. Decreased ERG expression also correlates with EndMT in tissues from patients with end-stage liver fibrosis. These studies identify a pathogenic mechanism where loss of ERG causes endothelial-dependent liver fibrogenesis via regulation of SMAD2/3. Moreover, ERG represents a promising candidate biomarker for assessing EndMT in liver disease.

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Section: Resultsmentioning

confidence: 99%

Dynamic regulation of canonical TGFβ signalling by endothelial transcription factor ERG protects from liver fibrogenesis

et al. 2017

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“…To minimize EndMT, remove contaminating cells that may act as a source TGFβ, keep cultures at high densities, and maintain high concentrations of bFGF in the media at all times. As we have shown recently, bFGF is essential to preserve EC specification as it antagonizes TGFβ-driven expression of smooth muscle actin 34 .…”

Section: Discussionmentioning

confidence: 81%

Isolation and Culture Expansion of Tumor-specific Endothelial Cells

Xiao

McCann

Dudley

2015

JoVE

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Freshly isolated tumor-specific endothelial cells (TEC) can be used to explore molecular mechanisms of tumor angiogenesis and serve as an in vitro model for developing new angiogenesis inhibitors for cancer. However, long-term in vitro expansion of murine endothelial cells is challenging due to phenotypic drift in culture (endothelial-to-mesenchymal transition) and contamination with non-endothelial cells (EC). This is especially true for TEC which are readily out competed by co-purified fibroblasts or tumor cells in culture. Here, a high fidelity isolation method that takes advantage of immunomagnetic enrichment coupled with colony selection and in vitro expansion is described. This approach generates pure endothelial cell fractions that are entirely free of contaminating stromal or tumor cells. It is also shown that lineage-traced Cdh5cre:ZsGreenl/s/l reporter mice, used with the protocol described herein, are a valuable tool to verify cell purity as the isolated EC colonies from these mice show durable and brilliant ZsGreen fluorescence in culture.

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“…; Xiao et al . ). We also found that TNF‐α induces the expression of miR‐31 and enhances EndMT and EndMT‐SP in MS‐1 cells (Fig.…”

Section: Discussionmentioning

confidence: 97%

MicroRNA‐31 is a positive modulator of endothelial–mesenchymal transition and associated secretory phenotype induced by TGF‐β

et al. 2015

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Transforming growth factor‐β (TGF‐β) plays central roles in endothelial–mesenchymal transition (EndMT) involved in development and pathogenesis. Although EndMT and epithelial–mesenchymal transition are similar processes, roles of microRNAs in EndMT are largely unknown. Here, we report that constitutively active microRNA‐31 (miR‐31) is a positive regulator of TGF‐β‐induced EndMT. Although the expression is not induced by TGF‐β, miR‐31 is required for induction of mesenchymal genes including α‐SMA, actin reorganization and MRTF‐A activation during EndMT. We identified VAV3, a regulator of actin remodeling and MRTF‐A activity, as a miR‐31 target. Global transcriptome analysis further showed that miR‐31 positively regulates EndMT‐associated unique secretory phenotype (EndMT‐SP) characterized by induction of multiple inflammatory chemokines and cytokines including CCL17, CX3CL1, CXCL16, IL‐6 and Angptl2. As a mechanism for this phenomenon, TGF‐β and miR‐31 suppress Stk40, a negative regulator of NF‐κB pathway. Interestingly, TGF‐β induces alternative polyadenylation (APA)‐coupled miR‐31‐dependent Stk40 suppression without concomitant miR‐31 induction, and APA‐mediated exclusion of internal poly(A) sequence in Stk40 3′UTR enhances target efficiency of Stk40. Finally, miR‐31 functions as a molecular hub to integrate TGF‐β and TNF‐α signaling to enhance EndMT. These data confirm that constitutively active microRNAs, as well as inducible microRNAs, serve as phenotypic modifiers interconnected with transcriptome dynamics during EndMT.

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Tumor Endothelial Cells with Distinct Patterns of TGFβ-Driven Endothelial-to-Mesenchymal Transition

Cited by 63 publications

References 49 publications

Dynamic regulation of canonical TGFβ signalling by endothelial transcription factor ERG protects from liver fibrogenesis

Dynamic regulation of canonical TGFβ signalling by endothelial transcription factor ERG protects from liver fibrogenesis

Isolation and Culture Expansion of Tumor-specific Endothelial Cells

MicroRNA‐31 is a positive modulator of endothelial–mesenchymal transition and associated secretory phenotype induced by TGF‐β

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