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RIPK3 modulates growth factor receptor expression in endothelial cells to support angiogenesis

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Abstract

Receptor-interacting protein kinase 3 (RIPK3) is a multifunctional intracellular protein that was first recognized as an important component of the necroptosis programmed cell death pathway. RIPK3 is also highly expressed in non-necroptotic murine embryonic endothelial cells (ECs) during vascular development, indicating its potential contribution to angiogenesis. To test this hypothesis, we generated mice lacking endothelial RIPK3 and found non-lethal embryonic and perinatal angiogenesis defects in multiple vascular beds. Our in vitro data indicate that RIPK3 supports angiogenesis by regulating growth factor receptor degradation in ECs. We found that RIPK3 interacted with the membrane trafficking protein myoferlin to sustain expression of vascular endothelial growth factor receptor 2 (VEGFR2) in cultured ECs following vascular endothelial growth factor A (VEGFA) stimulation. Restoration of myoferlin, which was diminished after RIPK3 knockdown, rescued decreased VEGFR2 expression and vascular sprouting in RIPK3-deficient ECs after VEGFA treatment. In addition, we found that RIPK3 modulated expression of genes involved in endothelial identity by inhibiting ERK signaling independently of growth factor receptor turnover. Altogether, our data reveal unexpected non-necroptotic roles for RIPK3 in ECs and evidence that RIPK3 promotes developmental angiogenesis in vivo.

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Acknowledgements

We thank Jun Xie for assistance with mouse colony maintenance, Kate Wheeler for technical help with this project, and Griffin lab members for helpful discussions. We also thank Steve Hartson (Oklahoma State University Protein Resource Core Facility) for assistance with LC-MS/MS. This work was supported by NIH grants R35HLI44605 (to C.T.G.) and P30GM114731 and by an AHA Predoctoral Fellowship #19PRE34380708 (to S.G.).

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  1. Siqi Gao

    Present address: Department of Medicine and Cardiovascular Institute, University of Pennsylvania, Philadelphia, PA, 19104, USA

Authors and Affiliations

  1. Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, 825 NE 13th Street, Oklahoma City, OK, 73104, USA

    Siqi Gao & Courtney T. Griffin

  2. Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104, USA

    Siqi Gao & Courtney T. Griffin

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Conceptualization, S.G and C.T.G.; Investigation, S.G.; Writing, S.G. and C.T.G.; Funding acquisition, C.T.G. and S.G.

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Correspondence to Courtney T. Griffin.

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All experimental animal protocols were approved by the Institutional Animal Care and Use Committee at the Oklahoma Medical Research Foundation.

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Gao, S., Griffin, C.T. RIPK3 modulates growth factor receptor expression in endothelial cells to support angiogenesis. Angiogenesis 24, 519–531 (2021). https://doi.org/10.1007/s10456-020-09763-5

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