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Capillary morphogenesis gene 2 (CMG2) mediates growth factor-induced angiogenesis by regulating endothelial cell chemotaxis

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Abstract

Anthrax protective antigen (PA) is a potent inhibitor of pathological angiogenesis with an unknown mechanism. In anthrax intoxication, PA interacts with capillary morphogenesis gene 2 (CMG2) and tumor endothelial marker 8 (TEM8). Here, we show that CMG2 mediates the antiangiogenic effects of PA and is required for growth-factor-induced chemotaxis. Using specific inhibitors of CMG2 and TEM8 interaction with natural ligand, as well as mice with the CMG2 or TEM8 transmembrane and intracellular domains disrupted, we demonstrate that inhibiting CMG2, but not TEM8 reduces growth-factor-induced angiogenesis in the cornea. Furthermore, the antiangiogenic effect of PA was abolished when the CMG2, but not the TEM8, gene was disrupted. Binding experiments demonstrated a broad ligand specificity for CMG2 among extracellular matrix (ECM) proteins. Ex vivo experiments demonstrated that CMG2 (but not TEM8) is required for PA activity in human dermal microvascular endothelial cell (HMVEC-d) network formation assays. Remarkably, blocking CMG2-ligand binding with PA or CRISPR knockout abolishes endothelial cell chemotaxis but not chemokinesis in microfluidic migration assays. These effects are phenocopied by Rho inhibition. Because CMG2 mediates the chemotactic response of endothelial cells to peptide growth factors in an ECM-dependent fashion, CMG2 is well-placed to integrate growth factor and ECM signals. Thus, CMG2 targeting is a novel way to inhibit angiogenesis.

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The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

The CMG2-Fc and TEM8-Fc fusion proteins were a kind gift of Planet Biotechnology. The SR-8F7/C7 anti-CMG2 antibody and associated data were a kind gift of Gisou van der Goot and Laurence Abrami. The L2 anti-TEM8 antibody was a kind gift of Brad St. Croix. Funding was provided by the NEI (R01EY018829), NINDS (R21NS059411), and DOD CDMRP (W81XWH-08-1-0710).

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    Authors and Affiliations

    1. Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT, 84602, USA

      Tsz-Ming Tsang, Sai Lun Lee, Samuel Garrard, Cody Roberts, Jessie Payne, Andrew Jensen, P. Christine Ackroyd & Kenneth A. Christensen

    2. Vascular Biology Program, Department of Surgery, Boston Children’s Hospital, Harvard Medical School, 11.211 Karp Family Research Bldg., 300 Longwood Ave., Boston, MA, 02115, USA

      Lorna M. Cryan, Jessica Stiles, Lauren Bazinet, Samuel Garrard, Erika Madrian & Michael S. Rogers

    3. Department of Medicine, West Palm Beach VA Medical Center, 7305 N Military Trail, West Palm Beach, FL, 33410, USA

      Arthur E. Frankel

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    Cryan, L.M., Tsang, TM., Stiles, J. et al. Capillary morphogenesis gene 2 (CMG2) mediates growth factor-induced angiogenesis by regulating endothelial cell chemotaxis. Angiogenesis 25, 397–410 (2022). https://doi.org/10.1007/s10456-022-09833-w

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