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In vivo delivery of the caveolin-1 scaffolding domain inhibits nitric oxide synthesis and reduces inflammation

Nature Medicine volume 6, pages 1362–1367 (2000)Cite this article

Abstract

Caveolin-1, the primary coat protein of caveolae, has been implicated as a regulator of signal transduction through binding of its “scaffolding domain” to key signaling molecules. However, the physiological importance of caveolin-1 in regulating signaling has been difficult to distinguish from its traditional functions in caveolae assembly, transcytosis, and cholesterol transport. To directly address the importance of the caveolin scaffolding domain in vivo, we generated a chimeric peptide with a cellular internalization sequence fused to the caveolin-1 scaffolding domain (amino acids 82–101). The chimeric peptide was efficiently taken up into blood vessels and endothelial cells, resulting in selective inhibition of acetylcholine (Ach)-induced vasodilation and nitric oxide (NO) production, respectively. More importantly, systemic administration of the peptide to mice suppressed acute inflammation and vascular leak to the same extent as a glucocorticoid or an endothelial nitric oxide synthase (eNOS) inhibitor. These data imply that the caveolin-1 scaffolding domain can selectively regulate signal transduction to eNOS in endothelial cells and that small-molecule mimicry of this domain may provide a new therapeutic approach.

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Figure 1: Caveolin scaffolding domain peptide inhibits acetylcholine (Ach) -induced relaxation and potentiates phenylephrine (PE)-induced contraction of the isolated mouse aorta.
Figure 2: Evidence that eNOS is a primary biological target of the caveolin scaffolding domain peptide in intact blood vessels.
Figure 3: Biotinylated AP–caveolin peptide inhibits vascular relaxation and localizes to the endothelium and adventitia of mouse aorta.
Figure 4: Cellular uptake and inhibition of NO release by the AP–caveolin scaffolding domain peptide in cultured endothelial cells and eNOS-transfected cells.
Figure 5: AP–caveolin peptide inhibits edema formation and vascular leakage.

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Acknowledgements

The authors wish to thank J.K. Crawford and I. Laroche from the W.M. Keck biotechnology resource center at Yale University School of Medicine. This work is supported by grants from the National Institute of Health (HL57665, HL 61371 and HL 64793 to W.C.S., a grant-in-aid from the American Heart Association (National Grant to W.C.S.). W.C.S. is an established investigator of the American Heart Association. J-P.G. is supported in part by fellowships from the Heart and Stroke Foundation of Canada, Fonds pour la Formation de Chercheurs etl' Aida a la Recherche (FCAR), and from the Canadian Institutes of Health Research.

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  1. Mariarosaria Bucci and Jean-Philippe Gratton: M.B. and J-P.G contributed equally to this work.

Authors and Affiliations

  1. Department of Pharmacology, Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, 06536, Connecticut, USA

    Mariarosaria Bucci, Jean-Philippe Gratton, Radu Daniel Rudic, Lisette Acevedo & William C. Sessa

  2. Dipartimento di Farmacologia Sperimental, 49 via Domenico Montesano, Napoli, 80131, Italy

    Mariarosaria Bucci, Fiorentina Roviezzo & Giuseppe Cirino

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  1. Mariarosaria Bucci
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Correspondence to William C. Sessa.

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Bucci, M., Gratton, JP., Rudic, R. et al. In vivo delivery of the caveolin-1 scaffolding domain inhibits nitric oxide synthesis and reduces inflammation. Nat Med 6, 1362–1367 (2000). https://doi.org/10.1038/82176

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