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Activated protein C blocks p53-mediated apoptosis in ischemic human brain endothelium and is neuroprotective

Nature Medicine volume 9, pages 338–342 (2003)Cite this article

Abstract

Activated protein C (APC) is a systemic anti-coagulant and anti-inflammatory factor1,2,3. It reduces organ damage in animal models of sepsis, ischemic injury and stroke1,4,5 and substantially reduces mortality in patients with severe sepsis6. It was not known whether APC acts as a direct cell survival factor or whether its neuroprotective effect5,7 is secondary to its anti-coagulant and anti-inflammatory effects1,2,3. We report that APC directly prevents apoptosis in hypoxic human brain endothelium through transcriptionally dependent inhibition of tumor suppressor protein p53, normalization of the pro-apoptotic Bax/Bcl-2 ratio and reduction of caspase-3 signaling. These mechanisms are distinct from those involving upregulation of the genes encoding the anti-apoptotic Bcl-2 homolog A1 and inhibitor of apoptosis protein-1 (IAP-1) by APC in umbilical vein endothelial cells8,9. Cytoprotection of brain endothelium by APC in vitro required endothelial protein C receptor (EPCR) and protease-activated receptor-1 (PAR-1), as did its in vivo neuroprotective activity in a stroke model of mice with a severe deficiency of EPCR10. This is consistent with work showing the direct effects of APC on cultured cells via EPCR and PAR-1 (ref. 9). Moreover, the in vivo neuroprotective effects of low-dose mouse APC seemed to be independent of its anti-coagulant activity. Thus, APC protects the brain from ischemic injury by acting directly on brain cells.

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Figure 1: APC has anti-apoptotic activity in hypoxic human BECs.
Figure 2: APC blocks p53-dependent apoptosis in hypoxic human BECs.
Figure 3: In vivo neuroprotective effects of recombinant mouse APC during focal ischemic stroke in mice requires EPCR and PAR-1.

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Acknowledgements

This work was supported by US National Institutes of Health grant HL63290 and HL52246.

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Author notes

  1. Tong Cheng and Dong Liu: T.C. and D.L. contributed equally to this study.

Authors and Affiliations

  1. Department of Neurosurgery and Center for Aging and Developmental Biology, Division of Neurovascular Biology, Frank P. Smith Neurosurgical Research Laboratory, University of Rochester Medical Center, Rochester, New York, USA

    Tong Cheng, Dong Liu & Berislav V. Zlokovic

  2. Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California, USA

    John H. Griffin & José A. Fernández

  3. W.M. Keck Center for Transgene Research and Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana, USA

    Francis Castellino & Elliot D. Rosen

  4. Department of Immunology, Saga Medical School, Saga, Japan

    Kenji Fukudome

  5. Socratech Laboratories, Rochester, New York, USA

    Berislav V. Zlokovic

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Correspondence to Berislav V. Zlokovic.

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Competing interests

B.V.Z. has a financial relationship with Socratech, LLC, relative to his service in the capacity of Scientific Program Director. He receives consultation compensation, stock ownership and other equity interest. The relationship includes patent applications and may include trademarks, copyrights and/or licensing agreements. J.H.G. has a financial relationship with Socratech, LLC, as a member of their Scientific Advisory Board, with similar compensations and inclusions.

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Cheng, T., Liu, D., Griffin, J. et al. Activated protein C blocks p53-mediated apoptosis in ischemic human brain endothelium and is neuroprotective. Nat Med 9, 338–342 (2003). https://doi.org/10.1038/nm826

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