Summary
A large body of in vitro studies has helped to elucidate intracellular pathways that lead to mitogenic signaling in vascular smooth muscle (VSM) cells. However, a limitation of these studies is that they fail to test the in vivo physiological significance especially because VSM proliferation, in the forms of intimal hyperplasia and restenosis, is an important clinical problem. The recent advent of adenoviral gene transfer technology has made possible to test the in vivo effects of specific molecular modulations of intracellular signal transduction pathways on physiological responses. For example, in VSM, adenoviruses can be delivered to the vessel wall to determine a gene/protein’s role in proliferative responses to vascular injury. This technology, once standardized and rendered safe for human applications, will be the basis of gene therapy and molecular medicine. Several exemplary applications have now been generated in the vascular system, including the use of an adenovirus containing the carboxyl-terminus of the β-adrenergic receptor kinase (βARKct), which binds to the βγ-subunits of activated heterotrimeric G proteins (Gβγ), to study the in vivo role of Gβγ in VSM intimal hyperplasia after vascular injury that leads to restenosis.
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References
Marinissen, M. J. and Gutkind, J. S. (2001) G protein-coupled receptors and signaling networks: emerging paradigms. Trends Pharmacol. Sci. 22, 368–376.
Caron, M. G. and Lefkowitz, R. J. (1993) Catecholamine receptors: structure, function and regulation. Recent Prog. Horm. Res. 48, 277–290.
Clapman, D. E. and Neer, E. J. (1993) New roles for G protein, βγ dimers in transmembrane signaling. Nature 365, 403–406.
Smrcka, A. V. and Scott, J. K. (2002) Discovery of ligands for βγ subunits from phage-displayed peptide libraries. Methods Enzymol. 344, 557–576.
Inglese, J., Koch, W. J., Touhara, K., and Lefkowitz, R. J. (1995) Gβγ interactions with PH domains and Ras-MAPK signaling pathways. Trends Biochem. Sci. 20, 151–156.
Pierce, K. L., Luttrell, L. M., and Lefkowitz, R. J. (2001) New mechanisms in heptahelical receptor signaling to mitogen activated protein kinase cascades. Oncogene 20, 1532–1539.
Koch, W. J., Hawes, B. E., Allen, L. F., and Lefkowitz, R, J. (1994) Direct evidence that Gi-coupled receptor stimulation of mitogen-activaed protein kinase is mediated by Gβγ activation of p21ras. Proc. Natl. Acad. Sci. USA 91, 12,706–12,710.
Crespo, P., Xu, N., Simonds, W. S., and Gutkind, J, S. (1994) Ras-dependent activation of AP kinase pathway mediated by G protein-βγ subunits. Nature 369, 418–420.
Luttrell, L. M., van Biesen, T., Hawes, B. E., Koch, W. J., Touhara, K., and Lefkowitz, R. J. (1995) Gβγ subunits mediate mitogen-activated protein kinase activation by the tyrosinekinase insulin-like growth factor 1 receptor. J. Biol. Chem. 270, 16,495–16,498.
Koch, W. J., Inglese, J., Stone, W. C., and Lefkowitz, R. J. (1993) The binding site for the βγ subunits of heterotrimeric G proteins on the β-adrenergic receptor kinase. J. Biol. Chem. 268, 8256–8260.
Drazner, M. H., Peppel, K. C., Dyer, S., Grant, A. O., Koch, W. J., and Lefkowitz, R. J. (1997) Potentiation of β-adrenergic signaling by adenoviral-mediated gene transfer in adult rabbit ventricular myocytes. J. Clin. Invest. 99, 288–296.
Koch, W. J., Hawes, B. E., Inglese, J., Luttrell, L. M., and Lefkowitz, R. J. (1994) Cellular expression of the carboxyl terminus of a G protein-coupled receptor kinase attenuates Gβγ-mediated signaling. J. Biol. Chem. 269, 6193–6197.
Iaccarino, G., Smithwick, L. A., Lefkowitz, R. J., and Koch, W. J. (1999) Targeting Gβγ signaling in arterial vascular smooth muscle proliferation: A novel strategy to limit restenosis. Proc. Natl. Acad. Sci. USA 96, 3945–3950.
Clowes, A. W., Reidy, M. A., and Clowes, N. M. (1983) Kinetics of cellular proliferation after arterial injury. I. Smooth muscle growth in the absence of endothelium. Lab. Invest. 49, 327–337.
Davies, M. G. and Hagen, P. O. (1994) Pathobiology of intimal hyperplasia. Br. J. Surg. 81, 1254–1260.
Davies, M. G., Huynh, T. T. T., Fulton, G. J., Lefkowitz, R. J., Svendsen, E., Hagen, P. O., et al. (1998) G protein signaling and vein graft intimal hyperplasia: Reduction of intimal hyperplasia in vein grafts by a Gβγ inhibitor suggests a major role of G protein signaling in lesion development. Arterioscler. Thromb. Vasc. Biol. 18, 1275–1280.
Huynh, T. T. T., Iaccarino, G., Davies, M. G., Svendsen, E., Koch, W. J., and Hagen, P. O. (1998) Adenoviral mediated inhibition of Gβγ signaling limits the development of vein graft intimal hyperplasia. Surgery 124, 177–186.
Amalfitano, A., Hauser, M. A. Hu, H., Serra, D., Begy, C. R., and Chamberlain, J. S. (1998) Production and characterization of improved adenovirus vectors with the E1, E2b, and E3 genes deleted. J. Virol. 72, 926–933.
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Iaccarino, G., Koch, W.J. (2004). In Vivo Adenoviral-Mediated Gene Transfer of the βARKct to Study the Role of Gβγ in Arterial Restenosis. In: Smrcka, A.V. (eds) G Protein Signaling. Methods in Molecular Biology™, vol 237. Humana Press. https://doi.org/10.1385/1-59259-430-1:181
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DOI: https://doi.org/10.1385/1-59259-430-1:181
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