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Thrombospondin-4 mediates TGF-β-induced angiogenesis

Oncogene volume 36, pages 5189–5198 (2017)Cite this article

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Abstract

TGF-β is a multifunctional cytokine affecting many cell types and implicated in tissue remodeling processes. Due to its many functions and cell-specific effects, the consequences of TGF-β signaling are process-and stage-dependent, and it is not uncommon that TGF-β exerts distinct and sometimes opposing effects on a disease progression depending on the stage and on the pathological changes associated with the stage. The mechanisms underlying cell- and process-specific effects of TGF-β are poorly understood. We are describing a novel pathway that mediates induction of angiogenesis in response to TGF-β1. We found that in endothelial cells (EC) thrombospondin-4 (TSP-4), a secreted extracellular matrix (ECM) protein, is upregulated in response to TGF-β1 and mediates the effects of TGF-β1 on angiogenesis. Upregulation of TSP-4 does not require the synthesis of new protein, is not caused by decreased secretion of TSP-4, and is mediated by activation of SMAD3. Using Thbs4−/− mice and TSP-4 shRNA, we found that TSP-4 mediated pro-angiogenic functions in cultured EC and angiogenesis in vivo in response to TGF-β1. We observed~3-fold increases in tumor mass and levels of angiogenesis markers in animals injected with TGF-β1, and these effects did not occur in Thbs4−/− animals. Injections of an inhibitor of TGF-β1 signaling SB-431542 also decreased the weights of tumors and cancer angiogenesis. Our results from in vivo angiogenesis models and cultured EC document that TSP-4 mediates upregulation of angiogenesis by TGF-β1. Upregulation of pro-angiogenic TSP-4 and selective effects of TSP-4 on EC may contribute to stimulation of tumor growth by TGF-β despite the inhibition of cancer cell proliferation.

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Abbreviations

bFGF:

basic fibroblasts growth factor

DMSO:

dimethyl sulfoxide

ECM:

extracellular matrix

HDMEC:

human dermal microvascular endothelial cells

HUVEC:

human umbilical vein endothelial cells

IP:

intra-peritoneal

KO:

knock-out

MAEC:

mouse aortic endothelial cells

MLEC:

mouse lung endothelial cells

OCT:

OCT embedding cryoembedding Matrix

shRNA:

small hairpin RNA

VSMC:

vascular smooth muscle cells

vWF:

von Willebrand factor

WT:

wild type.

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Acknowledgements

This work was supported by NIH R01HL117216 (OS-A and EP) and NIH CA177771 (OS-A). Isolation of HUVECs was supported by UL1TR000439. HUVECs were provided by a grant awarded to Clinical and Translational Science Collaborative of Cleveland, a grant from the National Center for Advancing Translational Sciences (UL1TR000439) component of the National Institutes of Health, and National Institutes of Health Roadmap for Medical Research.

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Author notes

  1. S Muppala, R Xiao and I Krukovets: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Cardiology, Cleveland Clinic, Cleveland, OH, USA

    S Muppala, R Xiao, I Krukovets, D Verbovetsky, R Yendamuri, N Habib, E Plow & O Stenina-Adognravi

  2. Department of Integrative Medical Sciences, North Ohio Medical University, Rootstown, OH, USA

    P Raman

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  1. S Muppala
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Correspondence to O Stenina-Adognravi.

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Muppala, S., Xiao, R., Krukovets, I. et al. Thrombospondin-4 mediates TGF-β-induced angiogenesis. Oncogene 36, 5189–5198 (2017). https://doi.org/10.1038/onc.2017.140

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