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bioRxiv · 10.1101/2025.02.11.637707

CD47 prevents Rac-mediated phagocytosis through Vav1 dephosphorylation

Miller, W. D.·Mishra, A. K.·Sheedy, C. J.·Bond, A.·Gardner, B. M.·Montell, D. J.·Morrissey, M. A.
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Abstract

CD47 is expressed by viable cells to protect against phagocytosis. CD47 is recognized by SIRP, an inhibitory receptor expressed by macrophages and other myeloid cells. Activated SIRP recruits SHP-1 and SHP-2 phosphatases but the inhibitory signaling cascade downstream of these phosphatases is not clear. In this study, we used time lapse imaging to measure how CD47 impacts the kinetics of phagocytosis. We found that targets with IgG antibodies were primarily phagocytosed through a Rac-based reaching mechanism. Targets also containing CD47 were only phagocytosed through a less frequent Rho-based sinking mechanism. Hyperactivating Rac2 eliminated the suppressive effect of CD47, suggesting that CD47 prevents activation of Rac and reaching phagocytosis. During IgG-mediated phagocytosis, the tyrosine kinase Syk phosphorylates the GEF Vav, which then activates the GTPase Rac to drive F-actin rearrangement and target internalization. CD47 inhibited Vav1 phosphorylation without impacting Vav1 recruitment to the phagocytic synapse or Syk phosphorylation. Macrophages expressing a hyperactive Vav1 were no longer sensitive to CD47. Together this data suggests that Vav1 is a key target of the CD47 signaling pathway.

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Miller, W. D., Mishra, A. K., Sheedy, C. J., Bond, A., Gardner, B. M., Montell, D. J., Morrissey, M. A.. 2025-02-12. CD47 prevents Rac-mediated phagocytosis through Vav1 dephosphorylation. https://doi.org/10.1101/2025.02.11.637707

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