Abstract
Increased matrix metalloproteinase-12 (MMP-12) has been implicated in atherosclerosis and many other inflammatory processes. To define MMP-12 functions in vivo, we generated transgenic rabbits that expressed human (h) MMP-12 gene under the control of a macrophage-specific promoter, the human scavenger receptor promoter. Two transgenic founder rabbits were found to have hMMP-12 transgene integration by Southern blot analysis. hMMP-12 mRNA was expressed in peritoneal and alveolar macrophages, and in tissues enriched in macrophages in transgenic rabbits. High levels of hMMP-12 protein were detected in the conditioned media of cultured peritoneal and alveolar macrophages from transgenic rabbits. Zymography showed that hMMP-12 secreted from macrophages possessed enzymatic activity toward β-casein. To evaluate the expression of hMMP-12 in inflammatory sites, we used carrageenan-induced granulomas as an in vivo model for tissue macrophages and foam cells. Granuloma size in transgenic rabbits was significantly increased compared to that in control rabbits, and histological examination revealed that granulomas of transgenic rabbits were enriched in macrophages associated with increased hMMP-12 expression. We believe that this transgenic rabbit model with increased expression of hMMP-12 may become a useful model for further mechanistic studies of MMP-12 in inflammatory diseases and cancer invasion; it is also an ideal model for testing the in vivo action of MMP-12 inhibitors.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Banda MJ and Werb Z (1981) Mouse macrophage elastase. Purification and characterization as a metalloproteinase. Biochem J 193: 589–605.
Bell FP and Schaub RG (1989) Comparison of lipid accumulation and metabolism in carrageenan-induced granulomas to aorta and blood monocytes of normal and cholesterol-fed rabbits. Exp Mol Pathol 50: 327–336.
Bosze Z, Hiripi L, Carnwath JW and Niemann H (2003) The transgenic rabbit as model for human diseases and as a source of biologically active recombinant proteins. Transgenic Res 12: 541–553.
Curci JA, Liao S, Huffman MD, Shapiro SD and Thompson RW (1998) Expression and localization of macrophage elastase (matrix metalloproteinase-12) in abdominal aortic aneurysms. J Clin Invest 102: 1900–1910.
Escary JL, Choy HA, Reue K, Wang XP, Castellani LW, Glass CK, et al. (1999) Paradoxical effect on atherosclerosis of hormone-sensitive lipase overexpression in macrophages. J Lipid Res 40: 397–404.
Fan J, Yamada T, Tokunaga O and Watanabe T (1991) Alterations in the functional characteristics of macrophages induced by hypercholesterolemia. Virchows Arch B Cell Pathol Mol Pathol 61: 19–27.
Fan J, Challah M and Watanabe T (1999) Transgenic rabbit models for biomedical research: current status, basic methods and future perspectives. Pathol Int 49: 583–594.
Fan J and Watanabe T (2003) Transgenic rabbits as therapeutic protein bioreactors and human disease models. Pharmacol Ther 99: 261–282.
Feinberg MW, Jain MK, Werner F, Sibinga NE, Wiesel P, Wang H et al. (2000) Transforming growth factor-beta 1 inhibits cytokine-mediated induction of human metalloelastase in macrophages. J Biol Chem 275: 25766–25773.
Gronski TJ, Jr Martin RL, Kobayashi DK, Walsh BC, Holman MC, Huber M et al. (1997) Hydrolysis of a broad spectrum of extracellular matrix proteins by human macrophage elastase. J Biol Chem 272: 12189–12194.
Horvai A, Palinski W, Wu H, Moulton KS, Kalla K and Glass CK (1995) Scavenger receptor A gene regulatory elements target gene expression to macrophages and to foam cells of atherosclerotic lesions. Proc Natl Acad Sci USA 92: 5391–5395.
Ichikawa T, Unoki H, Sun H, Shimoyamada H, Marcovina S, Shikama H, et al. (2002) Lipoprotein(a) promotes smooth muscle cell proliferation and dedifferentiation in atherosclerotic lesions of human apo(a) transgenic rabbits. Am J Pathol 160: 227–236.
Kelley JL, Suenram CA, Valente AJ, Sprague EA, Rozek MM and Schwartz CJ (1985) Evolution of foam cells in subcutaneous rabbit carrageenan granulomas. II. Tissue and macrophage lipid composition. Am J Pathol 120: 391–401.
Lemaitre V, O'Byrne TK, Borczuk AC, Okada Y, Tall AR and D'Armiento J (2001) ApoE knockout mice expressing human matrix metalloproteinase-1 in macrophages have less advanced atherosclerosis. J Clin Invest 107: 1227–1234.
Major AS, Dove DE, Ishiguro H, Su YR, Brown AM, Liu L et al. (2001) Increased cholesterol efflux in apolipoprotein AI (ApoAI)-producing macrophages as a mechanism for reduced atherosclerosis in ApoAI(-/-) mice. Arterioscler Thromb Vasc Biol 21: 1790–1795.
Matsumoto S, Kobayashi T, Katoh M, Saito S, Ikeda Y, Kobori M et al. (1998) Expression and localization of matrix metalloproteinase-12 in the aorta of cholesterol-fed rabbits: relationship to lesion development. Am J Pathol 153: 109–119.
Schaub RG, Bree MP, Hayes LL, Rudd MA, Rabbani L, Loscalzo J et al. (1994). Recombinant human macrophage colony-stimulating factor reduces plasma cholesterol and carrageenan granuloma foam cell formation in Watanabe heritable hyperlipidemic rabbits. Arterioscler Thromb 14: 70–76.
Schwartz CJ, Ghidoni JJ, Kelley JL, Sprague EA, Valente AJ and Suenram CA (1985) Evolution of foam cells in subcutaneous rabbit carrageenan granulomas: I. Light-microscopic and ultrastructural study. Am J Pathol 118: 134–150.
Shapiro SD (1999) Diverse roles of macrophage matrix metalloproteinases in tissue destruction and tumor growth. Thromb Haemost 82: 846–849.
Shiomi M, Ito T, Yamada S, Kawashima S and Fan J (2003) Development of an animal model for spontaneous myocardial infarction (WHHLMI rabbits). Arterioscler Thromb Vasc Biol 23: 1239–1244.
Sun H, Unoki H, Wang X, Liang J, Ichikawa, T, Arai, Y et al. (2002) Lipoprotein(a) enhances advanced atherosclerosis and vascular calcification in WHHL transgenic rabbits expressing human apolipoprotein(a). J Biol Chem 277: 47486–47492.
Unoki H, Fan J and Watanabe T (1999) Low-density lipoproteins modulate endothelial cells to secrete endothelin-1 in a polarized pattern: a study using a culture model system simulating arterial intima. Cell Tissue Res 295: 89–99.
Vaalamo M, Kariniemi AL, Shapiro SD and Saarialho-Kere U (1999) Enhanced expression of human metalloelastase (MMP-12) in cutaneous granulomas and macrophage migration. J Invest Dermatol 112: 499–505.
Warner RL, Lewis CS, Beltran L, Younkin EM, Varani J and Johnson KJ (2001) The role of metalloelastase in immune complex-induced acute lung injury. Am J Pathol 158: 2139–2144.
Westermarck J and Kahari VM (1999) Regulation of matrix metalloproteinase expression in tumor invasion. FASEB J 13: 781–792.
Wu L, Fan J, Matsumoto S and Watanabe T (2000) Induction and regulation of matrix metalloproteinase-12 by cytokines and CD40 signaling in monocyte/macrophages. Biochem Biophys Res Commun 269: 808–815.
Wu L, Tanimoto A, Murata Y, Fan J, Sasaguri Y and Watanabe T (2001) Induction of human matrix metalloproteinase-12 gene transcriptional activity by GM-CSF requires the AP-1 binding site in human U937 monocytic cells. Biochem Biophys Res Commun 285: 300–307.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Fan, J., Wang, X., Wu, L. et al. Macrophage-Specific Overexpression of Human Matrix Metalloproteinase-12 in Transgenic Rabbits. Transgenic Res 13, 261–269 (2004). https://doi.org/10.1023/B:TRAG.0000034717.70729.61
Issue Date:
DOI: https://doi.org/10.1023/B:TRAG.0000034717.70729.61