Thrombin Induces Surface and Intracellular Secretion of Amyloid Precursor Protein from Human Endothelial Cells

John R. Ciallella; Helmer Figueiredo; Virginia Smith-Swintosky; Joseph P. McGillis

doi:10.1055/s-0037-1614537

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1999; 81(04): 630-637
DOI: 10.1055/s-0037-1614537

Rapid Communication

Schattauer GmbH

Thrombin Induces Surface and Intracellular Secretion of Amyloid Precursor Protein from Human Endothelial Cells

Authors

John R. Ciallella

¹From the Department of Microbiology and Immunology, KY, USA

²Sanders Brown Research Center on Aging, KY, USA
Helmer Figueiredo

³Department of Toxicology, University of Kentucky College of Medicine, Lexington, KY, USA
Virginia Smith-Swintosky^*

²Sanders Brown Research Center on Aging, KY, USA
Joseph P. McGillis

¹From the Department of Microbiology and Immunology, KY, USA

²Sanders Brown Research Center on Aging, KY, USA

Abstract

Summary

Thrombin, a major coagulant and inflammatory mediator, was shown to regulate amyloid precursor protein (APP) secretion. APP is the protein from which the amyloid beta peptide (Aß) is derived. Aß forms the core of vascular and cerebral plaques in Alzheimer’s disease (AD). In this study, human umbilical vein endothelial cells (HUVEC) were used to examine the effects of thrombin on APP expression. Cell supernatants from thrombin-treated HUVEC were immunoblotted to measure secreted APP. Thrombin-induced secretion of APP peaks at approximately 30 min post-treatment. Immunohistochemical analysis found that APP is not colocalized with or secreted through the same pathway as coagulation factor VIII. The secretion of APP is thrombin receptor-mediated, since it is inhibited by the thrombin antagonist N-Acetyl-D-Phe-Pro1Amido-4-Guanidino-Butyl-1-Boronic Acid. It also is induced by treatment with a calcium ionophore. Moreover, APP secretion is protein kinase C (PKC)-dependent because it is blocked by the PKC inhibitor bisindolylmaleimide. APP secretion also occurs from the cell surface, possibly through direct cleavage by thrombin. Immunoreactivity on the surface of HUVEC decreased after thrombin treatment but not after treatment with a non-proteolytic thrombin receptor activator. These data suggest that thrombin induces APP secretion through a PKC-dependent mechanism, as well as from the cell surface. Our results are consistent with thrombin playing a role in AD pathology.

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References

References
1 Fenton II JW, Ofosu FA, Brezniak DV, Hassouna I H. Understanding thrombin and hemostasis. Hematol Oncol Clin North Am 1993; 6: 1107-19.
2 Webb ML, Taylor DS, Molloy CJ. Effects of thrombin receptor activating peptide on phosphoinositide hydrolysis and protein kinase C activation in cultured rat aortic smooth muscle cells: Evidence for “tethered ligand” activation of smooth muscle cell thrombin receptors. Biochem Pharmacol 1993; 45: 1577-82.
3 Sugama Y, Tiruppathi C, Janakidevi K, Anderson TT, Fenton II JW, Malik AB. Thrombin-induced expression of endothelial P-selectin and intercellular adhesion molecule-1: A mechanism for stabilizing neutrophil adhesion. J Cell Biol 1992; 119: 935-44.
4 Levine J, Harlan JM, Harker LA, Joseph ML, Counts RB. Thrombin-mediated release of factor VIII from human umbilical vein endothelial cells in culture. Blood 1982; 60 (02) 531-4.
5 Igarashi K, Murai H, Asaka J. Proteolytic processing of amyloid b protein precursor (APP) by thrombin. Biochem. Biophys Res Comm 1992; 185: 1000-4.
6 Davis-Salinas J, Saporito-Irwin SM, Donovan FM, Cunningham DD, Van Nostrand WE. Thrombin receptor activation induces secretion and non-amyloidogenic processing of amyloid b-protein precursor. J Biol Chem 1994; 269: 22623-7.
7 Kosik KS. Alzheimer’s Disease: A cell biological perspective. Science 1992; 256: 780-3.
8 Sola C, Mengod G, Probst A, Palacios JM. Differential regional and cellular distribution of b-amyloid precursor protein messenger RNAs containing and lacking the Kunitz protease inhibitor domain in the brain of human, rat and mouse. Neurosci 1993; 53: 267-95.
9 Ciallella JR, Rangnekar VV, McGillis JP. Heat shock alters Alzheimer’s b-amyloid precursor protein expression in human endothelial cells. J Neurosci Res 1994; 37: 769-76.
10 Mattson MP, Cheng B, Culwell AR, Esch FS, lieberburg I, Rydel RE. Evidence for exitoprotective and intraneuronal calcium-regulating roles for secreted forms of the B-amyloid precursor protein. Neuron 1993; 10: 243-54.
11 Smith RP, Higuchi DA, Broze GJ. Platelet coagulation factor XIa-inhibitor, a form of Alzheimer amyloid precursor protein. Science 1990; 248: 1126-8.
12 Carney DH, Mann R, Redin WR, Pernia SD, Berry D, Heggers JP, Hayward PG, Robson MC, Christie J, Annable C, Fenton ll JW, Glenn KC. Enhancement of incisional wound healing and neovascularization in normal rats by thrombin and synthetic thrombin receptor-activating peptides. J Clin Invest 1992; 89: 1469-77.
13 Gimbrone , Jr. MA, Cotran RS, Folkman J. Human vascular endothelial cells in culture, growth and DNA synthesis. J Cell Biol 1974; 60: 673-84.
14 Carlson SL, McGillis JP. Modulation of leukocyte adhesion, migration, and homing by neurotransmitters and neuropeptides. Meth Neurosci 1995; 24: 335-54.
15 Maciag T, Ilsley J, Kelley PR, Forand R. An endothelial cell growth factor from bovine hypothalamus: Identification and partial characterization. Proc Natl Acad Sci USA 1979; 76: 5674-8.
16 Kettner C, Mersinger L, Knabb R. The selective inhibition of thrombin by peptides of boroarginine. J Biol Chem 1990; 265: 18289-97.
17 Lynch JJ, Ferro TJ, Blumenstock FA, Brockenauer AM, Malik AB. Increased endothelial albumin permeability mediated by protein kinase C activation. J Clin Invest 1990; 85: 1991-8.
18 Toullec D, Pianetti P, Coste H, Bellevergue P, Grand-Perret T, Ajakane M, Baudet V, Boissin P, Boursier E, Loriolle F, Duhamel L, Charon D, Kirilovsky J. The bisindolylmaleimide GF 109203X is a potent and selective inhibitor of protein kinase c. J Biol Chem 1991; 266: 15771-81.
19 Ledoux S, Rebai N, Dagenais A, Shaw IT, Nalbantoglu J, Sekaly RP, Cash-man NR. Amyloid precursor protein in peripheral mononuclear cells is up-regulated with cell activation. J Immunol 1993; 150: 5566-75.
20 Malone DG, Pierce JH, Falko JH, Metcalfe DD. Production of granulocyte-macrophage colony-stimulating factor by primary cultures of unstimulated rat microvascular endothelial cells. Blood 1988; 71: 684-9.
21 Springer TA. Adhesion receptors of the immune system. Nature 1990; 346: 425-33.
22 Haass C, Koo EH, Mellon A, Hung AY, Selkoe DJ. Targeting of cell-surface b-amyloid precursor protein to lysosomes: Alternative processing into amyloid-bearing fragments. Nature 1992; 357: 500-3.
23 Yamaguchi H, Yamazaki T, Lemere CA, Frosch MP, Selkoe DJ. Beta Amyloid is focally deposited within the outer basement membrane in the amyloid angiopathy of Alzheimer’s Disease. Am J Pathol 1992; 141: 249-59.
24 Buxbaum JD, Oishi M, Chen I H, Pinkas-Kramarski R, Jaffe EA, Gandy SE, Greengard P. Cholinergic agonists and interleukin 1 regulate processing and secretion of the Alzheimer b/A4 amyloid protein precursor. Proc Natl Acad Sci USA 1992; 89: 10075-8.
25 Goldgaber D, Harris HW, Hla T, Maciag T, Donnelly RJ, Jacobsen JS, Vitek MP, Gajdusek C. Interleukin-1 regulates synthesis of amyloid b-protein precursor mRNA in human endothelial cells. Proc Nat. Acad Sci 1989; 86: 7606-10.
26 Kinouchi T, Sorimachi H, Maruyama K, Mizuno K, Ohno S, Ishiura S, Suzuki K. Conventional protein kinase C (PKC)-a and a novel PKCÎ, but not-d, increase the secretion of an N-terminal fragment of Alzheimer’s disease amyloid precursor protein from PKC cDNA transfected 3Y1 fibroblasts. FEBS Lett 1995; 364: 203-6.
27 Miyazaki K, Hasegawa M, Funahashi K, Umeda M. A metalloproteinase inhibitor domain in Alzheimer amyloid protein precursor. Nature 1993; 362: 839-41.
28 Scott GK. Proteinases and proteinase inhibitors as modulators of animal cell growth. Comp Biochem Pysiol 1992; 103b: 785-93.
29 Akiyama H, Ikeda K, Kondo H, McGeer PL. Thrombin accumulation in brains of patients with Alzheimer’s disease. Neurosci Lett 1992; 146: 152-4.
30 Davies TA, Fine RE, Johnson RJ, Levesque CA, Rathbun WH, Seetoo KF, Smith SJ, Strohmeier G, Volicer L, Delva L, Simons ER. Non-age related differences in thrombin responses by platelets from male patients with advanced Alzheimer’s disease. Bioc Biophys Res Comm 1993; 194: 537-43.
31 McKinney M, Snider RM, Richelson E. Thrombin binding to human brain and spinal cord. Mayo Clin Proc 1983; 58: 829-31.
32 Suidan HS, Stone SR, Hemmings BR, Monard D. Thrombin causes neurite retraction in neuronal cells through activation of cell surface receptors. Neuron 1992; 8: 363-75.