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Lysosomal Cholesterol Accumulation Inhibits Subsequent Hydrolysis of Lipoprotein Cholesteryl Ester

Published online by Cambridge University Press:  03 March 2008

W. Gray Jerome ,
Brian E. Cox ,
Evelyn E. Griffin  and
Jody C. Ullery
W. Gray Jerome
Affiliation:
Department of Pathology, Vanderbilt University School of Medicine, South Nashville, TN 37232-2561, USA Department of Cancer Biology, Vanderbilt University School of Medicine, South Nashville, TN 37232-2561, USA
Brian E. Cox
Affiliation:
Department of Pathology, Vanderbilt University School of Medicine, South Nashville, TN 37232-2561, USA
Evelyn E. Griffin
Affiliation:
Department of Pathology, Vanderbilt University School of Medicine, South Nashville, TN 37232-2561, USA
Jody C. Ullery
Affiliation:
Department of Pathology, Vanderbilt University School of Medicine, South Nashville, TN 37232-2561, USA
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Abstract

Human macrophages incubated for prolonged periods with mildly oxidized LDL (oxLDL) or cholesteryl ester-rich lipid dispersions (DISP) accumulate free and esterified cholesterol within large, swollen lysosomes similar to those in foam cells of atherosclerosis. The cholesteryl ester (CE) accumulation is, in part, the result of inhibition of lysosomal hydrolysis due to increased lysosomal pH mediated by excessive lysosomal free cholesterol (FC). To determine if the inhibition of hydrolysis was long lived and further define the extent of the lysosomal defect, we incubated THP-1 macrophages with oxLDL or DISP to produce lysosome sterol engorgement and then chased with acetylated LDL (acLDL). Unlike oxLDL or DISP, CE from acLDL normally is hydrolyzed rapidly. Three days of incubation with oxLDL or DISP produced an excess of CE in lipid-engorged lysosomes, indicative of inhibition. After prolonged oxLDL or DISP pretreatment, subsequent hydrolysis of acLDL CE was inhibited. Coincident with the inhibition, the lipid-engorged lysosomes failed to maintain an acidic pH during both the initial pretreatment and subsequent acLDL incubation. This indicates that the alterations in lysosomes were general, long lived, and affected subsequent lipoprotein metabolism. This same phenomenon, occurring within atherosclerotic foam cells, could significantly affect lesion progression.

Keywords

macrophageatherosclerosisfoam cellcholesterollysosomelipoprotein metabolism
Type
Research Article
Information
Microscopy and Microanalysis , Volume 14 , Issue 2 , April 2008 , pp. 138 - 149
Copyright
© 2008 Microscopy Society of America

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References

REFERENCES

Antonov, A., Munn, D., Kolodgie, F., Virmani, R. & Gerrity, R.G. (1997). Aortic endothelial cells regulate proliferation of human monoctyes in vitro via a mechanism synergistic with macrophage colony-stimulating factor. J Clin Invest 99, 2867–2876.Google Scholar
Auwerx, J. & Schoonjans, K. (1991). New insights into apolipoprotein B and low density lipoprotein physiology; implications for atherosclerosis. Acta Clin Belg 46, 355–358.Google Scholar
Barakat, H.A. & St. Clair, R.W. (1985). Characterization of plasma lipoproteins of grain- and cholesterol-fed white carneau and show racer pigeons. J Lipid Res 26, 1252–1268.Google Scholar
Basu, S.K., Goldstein, J.L., Anderson, R. & Brown, M.S. (1976). Degradation of cationized low density lipoprotein and regulation of cholesterol metabolism in homozygous hypercholesterolemia fibroblasts. Proc Natl Acad Sci USA 73, 3178–3182.Google Scholar
Bligh, E.G. & Dyer, W.J. (1959). A rapid method for total lipid extraction and purification. Protein measurement with the Folin phenol reagent. Can J Biochem Physiol 37, 911–917.Google Scholar
Brasaemle, D.L., Barber, T., Kimmel, A.R. & Londos, C. (1997). Post-translational regulation of perilipin expression. J Biol Chem 272, 9378–9387.Google Scholar
Brown, A., Mander, E., Gelissen, I., Kritharides, L., Dean, R. & Jessup, W. (2000). Cholesterol and oxysterol metabolism and subcellular distribution in macrophage foam cells: Accumulation of oxidized esters in lysosomes. J Lipid Res 41, 226–236.Google Scholar
Brown, M., Dana, S. & Goldstein, J. (1975). Receptor-dependent hydrolysis of cholesteryl esters contained in plasma low density lipoprotein. Proc Natl Acad Sci USA 72, 2925–2929.Google Scholar
Brown, M., Goldstein, J., Krieger, M., Ho, Y. & Anderson, R. (1979). Reversible accumulation of cholesteryl esters in macrophages incubated with acetylated lipoproteins. J Cell Biol 82, 597–613.Google Scholar
Cox, B., Griffin, E., Ullery, J. & Jerome, W. (2007). Effects of cellular cholesterol loading on macrophage foam cell lysosome acidification. J Lipid Res 48, 1012–1021.Google Scholar
Crider, B. & Xie, X.-S. (2003). Characterization of the functional coupling of bovine brain vacuolar-type H+ translocating ATPase: Effect of divalent cations, phospholipids, and subunit H (SFD). J Biol Chem 278, 44281–44288.Google Scholar
De Duve, C., De Barsy, T., Poole, B., Trouet, A., Tulkens, P. & Van Hoof, F. (1974). Lysosomotropic agents. Biochem Pharmacol 23, 2495–2531.Google Scholar
Dhaliwal, B. & Steinbrecher, U. (2000). Cholesterol delivered to macrophages by oxidized low density lipoprotein is sequestered in lysosomes and fails to efflux normally. J Lipid Res 41, 1658–1665.Google Scholar
Diwu, Z., Chen, C.-S., Zhang, C., Klaubert, D. & Haugland, R.P. (1999). A novel acidotropic pH indicator and its potential application in labeling acidic organelles of live cells. Chem Biol 6, 411–418.Google Scholar
Esterbauer, H., Rotheneder, M., Striegel, G., Waeg, G., Ashy, A., Sattler, W. & Jurgens, G. (1989). Vitamin E and other lipophilic antioxidants protect LDL against oxidation. Fat Sci Technol 8, 316–324.Google Scholar
Frens, G. (1973). Controlled nucleation for the regulation of particle size in monodisperse gold suspensions. Nat Phys Sci 241, 20–22.Google Scholar
Grabe, M. & Oster, G. (2001). Regulation of organelle acidity. J Gen Physiol 117, 329–343.Google Scholar
Greenspan, P. & Fowler, S. (1985). Spectrofluorometric studies of the lipid probe, nile red. J Lipid Res 26, 781–789.Google Scholar
Greenspan, P., Mayer, E. & Fowler, S. (1985). Nile red: A selective fluorescent stain for intracellular lipid droplets. J Cell Biol 100, 965–973.Google Scholar
Griffin, E., Ullery, J., Cox, B. & Jerome, W.G. (2005). Aggregated LDL and lipid dispersions induce lysosomal cholesteryl ester accumulation in macrophage foam cells. J Lipid Res 46, 2052–2060.Google Scholar
Guyton, J. & Klemp, K. (1994). Development of the atherosclerotic core region. Chemical and ultrastructural analysis of microdissected atherosclerotic lesions from human aorta. Arterioscler Thromb 14, 1305–1314.Google Scholar
Handley, D.A., Arbeeny, C.M., Witte, L.D. & Chien, S. (1981). Colloidal gold-low density lipoprotein conjugates as membrane receptor probes. Proc Natl Acad Sci USA 78, 368–371.Google Scholar
Hoff, H. & Morton, R. (1985). Lipoproteins containing apo B extracted from human aortas: Structure and function. Ann NY Acad Sci 454, 183–194.Google Scholar
Hoppe, G., O'Neil, J. & Hoff, H.F. (1994). Inactivation of lysosomal proteases by oxidized low density lipoprotein is partially responsible for its poor degradation by mouse peritoneal macrophages. J Clin Invest 94, 1506–1512.Google Scholar
Jerome, W., Cash, C., Webber, R., Horton, R. & Yancey, P. (1998). Lysosomal lipid accumulation from oxidized low density lipoprotein is correlated with hypertrophy of the Golgi apparatus and trans-Golgi network. J Lipid Res 39, 1362–1371.Google Scholar
Jerome, W.G. (2006). Advanced atherosclerotic foam cell formation has features of an acquired lysosomal storage disorder. Rejuvenation Res 9, 245–255.Google Scholar
Jerome, W.G. & Lewis, J.C. (1985). Early atherogenesis in White Carneau pigeons, II: Ultrastructural and cytochemical observations. Am J Pathol 119, 210–222.Google Scholar
Jerome, W.G. & Lewis, J.C. (1997). Cellular dynamics in early atherosclerotic lesion progression in White Carneau pigeons. Spatial and temporal analysis of monocyte and smooth muscle invasion of the intima. Arterioscler Thromb Vasc Biol 17, 654–664.Google Scholar
Jerome, W.G. & Yancey, P.G. (2003). The role of microscopy in understanding atherosclerotic lysosomal lipid metabolism. Microsc Microanal 9, 54–67.Google Scholar
Kirk, R.E. (1968). Experimental design: Procedures for the behavioral sciences. Belmont, CA: Brooks/Cole Publishing Co.
Klansek, J.J., Yancey, P.G., St. Clair, R.W., Fischer, R.T., Johnson, W.J. & Glick, J.M. (1995). Cholesterol quantitation by GLC: Artifactual formation of short-chain steryl esters. J Lipid Res 36, 2261–2266.Google Scholar
Kritharides, L., Upston, J., Jessup, W. & Dean, R. (1998). Accumulation and metabolism of low density lipoprotein-derived cholesteryl linoleate hydroperoxide and hydroxide by macrophages. J Lipid Res 39, 2394–2405.Google Scholar
Kruth, H. (1984). Localization of unesterified cholesterol in human atherosclerotic lesions. Demonstration of filipin-positive, oil-red-O-negative particles. Am J Pathol 114, 201–208.Google Scholar
Lin, H.-J., Herman, P., Kang, J. & Lakowicz, J. (2001). Fluorescence lifetime characterization of novel low-pH probes. Anal Biochem 294, 118–125.Google Scholar
Lougheed, M., Zhang, H. & Steinbrecher, U. (1991). Oxidized low density lipoprotein is resistant to cathepsins and accumulates within macrophages. J Biol Chem 266, 14519–14525.Google Scholar
Lowry, O.H., Rosebrough, N.J., Farr, A.L. & Randall, R.J. (1951). Protein measurement with the Folin phenol reagent. J Biol Chem 193, 265–275.Google Scholar
Mahlberg, F., Glick, J.M., Jerome, W.G. & Rothblat, G.H. (1990). Metabolism of cholesteryl ester lipid droplets in a J774 macrophage foam cell model. Biochim Biophys Acta 1045, 291–298.Google Scholar
Maor, I. & Aviram, M. (1994). Oxidized low density lipoprotein leads to macrophage accumulation of unesterified cholesterol as a result of lysosomal trapping of the lipoprotein hydrolyzed cholesteryl ester. J Lipid Res 35, 803–819.Google Scholar
McGookey, D. & Anderson, R. (1983). Morphological characterization of the cholesteryl ester cycle in cultured mouse macrophage foam cells. J Cell Biol 97, 1156–1168.Google Scholar
Munn, D. & Armstrong, E. (1993). Cytokine regulation of human monocyte differentiation in vitro: The tumor-cytotoxic phenotype induced by macrophage colony-stimulating factor is developmentally regulated by gamma-interferon. Cancer Res 53, 2603–2613.Google Scholar
O'Neil, J., Hoppe, G. & Hoff, H. (2003). Phospholipids in oxidized low density lipoproteins perturb the ability of macrophages to degrade internalized macromolecules and reduce intracellular cathepsin B activity. Atherosclerosis 169, 215–224.Google Scholar
Peters, T.J. & De Duve, C. (1974). Lysosomes of the arterial wall II. Subcellular fractionation of aortic cells from rabbits with experimental atheroscleroma. Exp Mol Pathol 20, 228–256.Google Scholar
Roma, P., Bernini, F., Fogliatto, R., Bertulli, S.M., Negri, S., Fumagalli, R. & Catapano, A.L. (1992). Defective catabolism of oxidized LDL by J774 murine macrophages. J Lipid Res 33, 819–829.Google Scholar
Rudel, L., Lee, R. & Cockman, T. (2001). Acyl coenzyme A:cholesterol acyltransferase types 1 and 2: Structure and function in atherosclerosis. Curr Opin Lipidol 12, 121–127.Google Scholar
Smith, E., Slater, R. & Chiu, P. (1968). The lipids in raised fatty and fibrous lesions in human aortas. A comparison of changes at different stages of development. J Atheroscler Res 8, 399–419.Google Scholar
Soyombo, A., Tjon-Kon-Sang, S., Rbaibi, Y., Bashllari, E., Bisceglia, J., Muallem, S. & Kiselyov, K. (2006). TRP-ML1 regulates lysosomal pH and acidic lysosomal lipid hydrolytic activity. J Biol Chem 281, 7294–7301.Google Scholar
Weibel, E.R., Kistler, G.S. & Scherle, W.F. (1966). Practical stereological methods for morphometric cytology. J Cell Biol 30, 23–38.Google Scholar
Yancey, P., Rothblat, G., Davidson, W., Kilsdonk, E., Atger, V. & de la Llera Moya, M. (1995). Cholesterol efflux from cells in culture. In Atherosclerosis X, Woodford, F., Davignon, J., Sniderman, A. (Eds.), pp. 481–484. Amsterdam: Elsevier Science.
Yancey, P.G. & Jerome, W.G. (1998). Lysosomal sequestration of free and esterified cholesterol from oxidized low density lipoprotein in macrophages of different species. J Lipid Res 39, 1349–1361.Google Scholar
Yancey, P.G. & Jerome, W.G. (2001). Lysosomal cholesterol derived from mildly oxidized low density lipoprotein is resistant to efflux. J Lipid Res 42, 317–327.Google Scholar
Yancey, P.G., Miles, S., Schwegel, J. & Jerome, W.G. (2002). Uptake and trafficking of mildly oxidized LDL and acetylated LDL in THP-1 cells does not explain the differences in lysosomal metabolism of these two lipoproteins. Microsc Microanal 8, 81–93.Google Scholar
Yancey, P.G., Rodrigueza, W.V., Kilsdonk, E.P.C., Stoudt, G.W., Johnson, W.J., Phillips, M.C. & Rothblat, G.H. (1996). Cellular cholesterol efflux mediated by cyclodextrins. Demonstration of kinetic pools and mechanism of efflux. J Biol Chem 271, 16026–16034.Google Scholar
Yancey, P.G. & St. Clair, R.W. (1992). Cholesterol efflux is defective in macrophages from atherosclerosis-susceptible White Carneau pigeons relative to resistant Show Racer pigeons. Arterioscler Thromb 12, 1291–1304.Google Scholar