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mTORC2 mediates CXCL12-induced angiogenesis

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Abstract

The chemokine CXCL12, through its receptor CXCR4, positively regulates angiogenesis by promoting endothelial cell (EC) migration and tube formation. However, the relevant downstream signaling pathways in EC have not been defined. Similarly, the upstream activators of mTORC2 signaling in EC are also poorly defined. Here, we demonstrate for the first time that CXCL12 regulation of angiogenesis requires mTORC2 but not mTORC1. We find that CXCR4 signaling activates mTORC2 as indicated by phosphorylation of serine 473 on Akt and does so through a G-protein- and PI3K-dependent pathway. Significantly, independent disruption of the mTOR complexes by drugs or multiple independent siRNAs reveals that mTORC2, but not mTORC1, is required for microvascular sprouting in a 3D in vitro angiogenesis model. Importantly, in a mouse model, both tumor angiogenesis and tumor volume are significantly reduced only when mTORC2 is inhibited. Finally, 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3), which is a key regulator of glycolytic flux, is required for microvascular sprouting in vitro, and its expression is reduced in vivo when mTORC2 is targeted. Taken together, these findings identify mTORC2 as a critical signaling nexus downstream of CXCL12/CXCR4 that represents a potential link between mTORC2, metabolic regulation, and angiogenesis.

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Acknowledgments

We thank Nazilya Gasanova, Sarah M. Sukardi, and Kimberly Lim for their help in quantifying the immunohistochemical specimens; Dr. Kehui Wang of the Pathology Research Services Core at UCI for processing the tissue sections for analysis; and Dr. David Fruman from the Department of Molecular Biology and Biochemistry at UCI for his helpful discussions during the course of these experiments. This work was supported by the National Institutes of Health/National Cancer Institute Institutional Training Grant Fellowship T32CA009054 to M.E.Z and RO1 HL60067 to C.C.W.H. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Cancer Institute or the National Institutes of Health. C.C.W.H. receives support from the Chao Family Comprehensive Cancer Center through a National Cancer Institute Center Grant, P30A062203.

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Authors and Affiliations

  1. The Department of Molecular Biology and Biochemistry, University of California Irvine, 3219 McGaugh Hall, Mail Code: 3900, Irvine, CA, 92697, USA

    Mary E. Ziegler, Michaela M. S. Hatch, Nan Wu, Steven A. Muawad & Christopher C. W. Hughes

  2. The Department of Biomedical Engineering, University of California Irvine, Irvine, CA, 92697, USA

    Christopher C. W. Hughes

  3. The Edwards Lifesciences Center for Advanced Cardiovascular Technology, University of California Irvine, Irvine, CA, 92697, USA

    Christopher C. W. Hughes

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  1. Mary E. Ziegler
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  2. Michaela M. S. Hatch
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Correspondence to Christopher C. W. Hughes.

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Ziegler, M.E., Hatch, M.M.S., Wu, N. et al. mTORC2 mediates CXCL12-induced angiogenesis. Angiogenesis 19, 359–371 (2016). https://doi.org/10.1007/s10456-016-9509-6

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