Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Apolipoprotein A-I and lecithin:cholesterol acyltransferase transfer induce cholesterol unloading in complex atherosclerotic lesions

Gene Therapy volume 16, pages 757–765 (2009)Cite this article

Abstract

Plasma levels of high-density lipoprotein (HDL) cholesterol and its major apolipoprotein (apo), apo A-I, are inversely correlated with the incidence of ischemic cardiovascular diseases. Reverse cholesterol transport is likely the main mechanism underlying the atheroprotective effects of HDL. Here, we investigated whether increased HDL cholesterol following hepatocyte-directed adenoviral rabbit apo A-I (AdrA-I) or rabbit lecithin-cholesterol acyltransferase (LCAT) (AdrLCAT) transfer may induce cholesterol unloading in complex atherosclerotic lesions in heterozygous low-density lipoprotein receptor-deficient rabbits fed a 0.15% cholesterol diet for 420 days before and for 120 days after transfer. HDL cholesterol levels increased 2.0-fold (P<0.001) and 1.9-fold (P<0.001) in the 120 days after transfer with AdrA-I and AdrLCAT, respectively, compared to levels just before transfer whereas non-HDL cholesterol remained unchanged. Increased HDL cholesterol following AdrA-I and AdrLCAT transfer resulted in a 31% (P<0.05) reduction of the intima/media ratio in comparison with the control progression group. Compared to the baseline group killed after 420 days of cholesterol diet, AdrA-I and AdrLCAT transfer reduced the percentage of Oil Red O area 1.6-fold (P<0.001) and 1.4-fold (P<0.001), respectively. In conclusion, increased HDL cholesterol after AdrA-I and AdrLCAT transfer inhibits progression of atherosclerosis and induces cholesterol unloading in complex lesions in rabbits.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to the full article PDF.

USD 39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

Abbreviations

Apo:

apolipoprotein

LCAT:

lecithin:cholesterol acyltransferase

LDL:

low-density lipoproteins

HDL:

high-density lipoproteins

LDLr:

low-density lipoprotein receptor.

References

  1. Gordon DJ, Rifkind BM . High-density lipoprotein—the clinical implications of recent studies. N Eng J Med 1989; 321: 1311–1316.

    Article  CAS  Google Scholar 

  2. Rubin EM, Krauss RM, Spangler EA, Verstuyft JG, Clift SM . Inhibition of early atherogenesis in transgenic mice by human apolipoprotein AI. Nature 1991; 353: 265–267.

    Article  CAS  Google Scholar 

  3. Duverger N, Kruth H, Emmanuel F, Caillaud JM, Viglietta C, Castro G et al. Inhibition of atherosclerosis development in cholesterol-fed human apolipoprotein A-I-transgenic rabbits. Circulation 1996; 94: 713–717.

    Article  CAS  Google Scholar 

  4. Paszty C, Maeda N, Verstuyft J, Rubin EM . Apolipoprotein AI transgene corrects apolipoprotein E deficiency-induced atherosclerosis in mice. J Clin Invest 1994; 94: 899–903.

    Article  CAS  Google Scholar 

  5. Plump AS, Scott CJ, Breslow JL . Human apolipoprotein A-I gene expression increases high density lipoprotein and suppresses atherosclerosis in the apolipoprotein E-deficient mouse. Proc Natl Acad Sci USA 1994; 91: 9607–9611.

    Article  CAS  Google Scholar 

  6. Badimon JJ, Badimon L, Fuster V . Regression of atherosclerotic lesions by high density lipoprotein plasma fraction in the cholesterol-fed rabbit. J Clin Invest 1990; 85: 1234–1241.

    Article  CAS  Google Scholar 

  7. Tangirala RK, Tsukamoto K, Chun SH, Usher D, Pure E, Rader DJ . Regression of atherosclerosis induced by liver-directed gene transfer of apolipoprotein A-I in mice. Circulation 1999; 100: 1816–1822.

    Article  CAS  Google Scholar 

  8. Kosek AB, Durbin D, Jonas A . Binding affinity and reactivity of lecithin cholesterol acyltransferase with native lipoproteins. Biochem Biophys Res Commun 1999; 258: 548–551.

    Article  CAS  Google Scholar 

  9. Greeve J, Altkemper I, Dieterich JH, Greten H, Windler E . Apolipoprotein B mRNA editing in 12 different mammalian species: hepatic expression is reflected in low concentrations of apoB-containing plasma lipoproteins. J Lipid Res 1993; 34: 1367–1383.

    CAS  PubMed  Google Scholar 

  10. Nagashima M, McLean JW, Lawn RM . Cloning and mRNA tissue distribution of rabbit cholesteryl ester transfer protein. J Lipid Res 1988; 29: 1643–1649.

    CAS  PubMed  Google Scholar 

  11. Tall AR . Plasma cholesteryl ester transfer protein. J Lipid Res 1993; 34: 1255–1274.

    CAS  PubMed  Google Scholar 

  12. Barter P, Rye KA . Cholesteryl ester transfer protein: its role in plasma lipid transport. Clin Exp Pharmacol Physiol 1994; 21: 663–672.

    Article  CAS  Google Scholar 

  13. Tall A . Plasma lipid transfer proteins. Annu Rev Biochem 1995; 64: 235–257.

    Article  CAS  Google Scholar 

  14. Lievens J, Snoeys J, Vekemans K, Van Linthout S, de Zanger R, Collen D et al. The size of sinusoidal fenestrae is a critical determinant of hepatocyte transduction after adenoviral gene transfer. Gene Therapy 2004; 11: 1523–1531.

    Article  CAS  Google Scholar 

  15. Snoeys J, Lievens J, Wisse E, Jacobs F, Duimel H, Collen D et al. Species differences in transgene DNA uptake in hepatocytes after adenoviral transfer correlate with the size of endothelial fenestrae. Gene Therapy 2007; 14: 604–612.

    Article  CAS  Google Scholar 

  16. Wisse E, Jacobs F, Topal B, Frederik P, De Geest B . The size of endothelial fenestrae in human liver sinusoids: implications for hepatocyte-directed gene transfer. Gene Therapy 2008; 15: 1193–1199.

    Article  CAS  Google Scholar 

  17. De Geest B, Van Linthout S, Collen D . Sustained expression of human apo A-I following adenoviral gene transfer in mice. Gene Therapy 2001; 8: 121–127.

    Article  CAS  Google Scholar 

  18. De Geest BR, Van Linthout SA, Collen D . Humoral immune response in mice against a circulating antigen induced by adenoviral transfer is strictly dependent on expression in antigen-presenting cells. Blood 2003; 101: 2551–2556.

    Article  CAS  Google Scholar 

  19. Mingozzi F, Liu YL, Dobrzynski E, Kaufhold A, Liu JH, Wang Y et al. Induction of immune tolerance to coagulation factor IX antigen by in vivo hepatic gene transfer. J Clin Invest 2003; 111: 1347–1356.

    Article  CAS  Google Scholar 

  20. Van Linthout S, Collen D, De Geest B . Effect of promoters and enhancers on expression, transgene DNA persistence, and hepatotoxicity after adenoviral gene transfer of human apolipoprotein A-I. Hum Gene Ther 2002; 13: 829–840.

    Article  CAS  Google Scholar 

  21. Van Linthout S, Lusky M, Collen D, De Geest B . Persistent hepatic expression of human apo A-I after transfer with a helper-virus independent adenoviral vector. Gene Therapy 2002; 9: 1520–1528.

    Article  CAS  Google Scholar 

  22. Belalcazar LM, Merched A, Carr B, Oka K, Chen KH, Pastore L et al. Long-term stable expression of human apolipoprotein A-I mediated by helper-dependent adenovirus gene transfer inhibits atherosclerosis progression and remodels atherosclerotic plaques in a mouse model of familial hypercholesterolemia. Circulation 2003; 107: 2726–2732.

    Article  CAS  Google Scholar 

  23. Brousseau ME, Kauffman RD, Herderick EE, Demosky Jr SJ, Evans W, Marcovina S et al. LCAT modulates atherogenic plasma lipoproteins and the extent of atherosclerosis only in the presence of normal LDL receptors in transgenic rabbits. Arterioscler Thromb Vasc Biol 2000; 20: 450–458.

    Article  CAS  Google Scholar 

  24. Shore VG, Shore B, Hart RG . Changes in apolipoproteins and properties of rabbit very low density lipoproteins on induction of cholesteremia. Biochemistry 1974; 13: 1579–1585.

    Article  CAS  Google Scholar 

  25. Roth RI, Gaubatz JW, Gotto Jr AM, Patsch JR . Effect of cholesterol feeding on the distribution of plasma lipoproteins and on the metabolism of apolipoprotein E in the rabbit. J Lipid Res 1983; 24: 1–11.

    CAS  PubMed  Google Scholar 

  26. Chao YS, Yamin TT, Thompson GM, Kroon PA . Tissue-specific expression of genes encoding apolipoprotein E and apolipoprotein A-I in rabbits. J Biol Chem 1984; 259: 5306–5309.

    CAS  PubMed  Google Scholar 

  27. Sun YP, Lu NC, Parmley WW, Hollenbeck CB . Effects of cholesterol diets on vascular function and atherogenesis in rabbits. Proc Soc Exp Biol Med 2000; 224: 166–171.

    Article  CAS  Google Scholar 

  28. Kritchevsky D, Tepper SA, Vesselinovitch D, Wissler RW . Cholesterol vehicle in experimental atherosclerosis. 11. Peanut oil. Atherosclerosis 1971; 14: 53–64.

    Article  CAS  Google Scholar 

  29. Adams CW, Miller NE, Morgan RS, Rao SN . Lipoprotein levels and tissue lipids in fatty-fibrous atherosclerosis induced in rabbits by two years’ cholesterol feeding at a low level. Atherosclerosis 1982; 44: 1–8.

    Article  CAS  Google Scholar 

  30. Spagnoli LG, Orlandi A, Mauriello A, Santeusanio G, de Angelis C, Lucreziotti R et al. Aging and atherosclerosis in the rabbit. 1. Distribution, prevalence and morphology of atherosclerotic lesions. Atherosclerosis 1991; 89: 11–24.

    Article  CAS  Google Scholar 

  31. Bocan TM, Mueller SB, Uhlendorf PD, Ferguson E, Newton RS . Dietary and mechanically induced rabbit iliac-femoral atherosclerotic lesions: a chemical and morphologic evaluation. Exp Mol Pathol 1991; 54: 201–277.

    Article  CAS  Google Scholar 

  32. Lupu F, Danaricu I, Simionescu N . Development of intracellular lipid deposits in the lipid-laden cells of atherosclerotic lesions. A cytochemical and ultrastructural study. Atherosclerosis 1987; 67: 127–142.

    Article  CAS  Google Scholar 

  33. Daley SJ, Klemp KF, Guyton JR, Rogers KA . Cholesterol-fed and casein-fed rabbit models of atherosclerosis. Part 2: differing morphological severity of atherogenesis despite matched plasma cholesterol levels. Arterioscler Thromb 1994; 14: 105–141.

    Article  CAS  Google Scholar 

  34. Kolodgie FD, Katocs Jr AS, Largis EE, Wrenn SM, Cornhill JF, Herderick EE et al. Hypercholesterolemia in the rabbit induced by feeding graded amounts of low-level cholesterol. Methodological considerations regarding individual variability in response to dietary cholesterol and development of lesion type. Arterioscler Thromb Vasc Biol 1996; 16: 1454–1464.

    Article  CAS  Google Scholar 

  35. Atkinson JB, Hoover RL, Berry KK, Swift LL . Cholesterol-fed heterozygous Watanabe heritable hyperlipidemic rabbits: a new model for atherosclerosis. Atherosclerosis 1989; 78: 123–136.

    Article  CAS  Google Scholar 

  36. Hoeg JM, Santamarina-Fojo S, Berard AM, Cornhill JF, Herderick EE, Feldman SH et al. Overexpression of lecithin:cholesterol acyltransferase in transgenic rabbits prevents diet-induced atherosclerosis. Proc Natl Acad Sci USA 1996; 93: 11448–11453.

    Article  CAS  Google Scholar 

  37. Berard AM, Foger B, Remaley A, Shamburek R, Vaisman BL, Talley G et al. High plasma HDL concentrations associated with enhanced atherosclerosis in transgenic mice overexpressing lecithin-cholesteryl acyltransferase. Nat Med 1997; 3: 744–749.

    Article  CAS  Google Scholar 

  38. Mehlum A, Muri M, Hagve TA, Solberg LA, Prydz H . Mice overexpressing human lecithin:cholesterol acyltransferase are not protected against diet-induced atherosclerosis. APMIS 1997; 105: 861–868.

    Article  CAS  Google Scholar 

  39. Mehlum A, Gjernes E, Solberg LA, Hagve TA, Prydz H . Overexpression of human lecithin:cholesterol acyltransferase in mice offers no protection against diet-induced atherosclerosis. APMIS 2000; 108: 336–342.

    Article  CAS  Google Scholar 

  40. Furbee Jr JW, Parks JS . Transgenic overexpression of human lecithin:cholesterol acyltransferase (LCAT) in mice does not increase aortic cholesterol deposition. Atherosclerosis 2002; 165: 89–100.

    Article  CAS  Google Scholar 

  41. Kondo T, Watanabe Y . [A heritable hyperlipemic rabbit (author's translation)]. Jikken Dobutsu 1975; 24: 89–94.

    CAS  PubMed  Google Scholar 

  42. Watanabe Y . Serial inbreeding of rabbits with hereditary hyperlipidemia (WHHL- rabbit). Atherosclerosis 1980; 36: 261–268.

    Article  CAS  Google Scholar 

  43. Sun H, Usui S, Shiomi M, Watanabe T, Fan J . A rapid PCR method of genotyping LDL receptor mutations in WHHL rabbits. J Atheroscler Thromb 2002; 9: 145–148.

    Article  CAS  Google Scholar 

  44. Murata Y, Maeda E, Yoshino G, Kasuga M . Cloning of rabbit LCAT cDNA: increase in LCAT mRNA abundance in the liver of cholesterol-fed rabbits. J Lipid Res 1996; 37: 1616–1622.

    CAS  PubMed  Google Scholar 

  45. Pan TC, Hao QL, Yamin TT, Dai PH, Chen BS, Chen SL et al. Rabbit apolipoprotein A-I mRNA and gene. Evidence that rabbit apolipoprotein A-I is synthesized in the intestine but not in the liver. Eur J Biochem 1987; 170: 99–104.

    Article  CAS  Google Scholar 

  46. Feng Y, Jacobs F, Van Craeyveld E, Brunaud C, Snoeys J, Tjwa M et al. Human ApoA-I transfer attenuates transplant arteriosclerosis via enhanced incorporation of bone marrow-derived endothelial progenitor cells. Arterioscler Thromb Vasc Biol 2008; 28: 278–283.

    Article  CAS  Google Scholar 

  47. Chapman MJ, Goldstein S, Lagrange D, Laplaud PM . A density gradient ultracentrifugal procedure for the isolation of the major lipoprotein classes from human serum. J Lipid Res 1981; 22: 339–358.

    CAS  PubMed  Google Scholar 

  48. Jacobs F, Van Craeyveld E, Feng Y, Snoeys J, De Geest B . Adenoviral low density lipoprotein receptor attenuates progression of atherosclerosis and decreases tissue cholesterol levels in a murine model of familial hypercholesterolemia. Atherosclerosis 2008; 201: 289–297.

    Article  CAS  Google Scholar 

  49. Junqueira LC, Bignolas G, Brentani RR . Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections. Histochem J 1979; 11: 447–455.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by grants G.0564.05 and G.0533.08 of the Fonds voor Wetenschappelijk Onderzoek-Vlaanderen. The Center for Molecular and Vascular Biology is supported by the Excellentiefinanciering KU Leuven (EF/05/013). Eline Van Craeyveld is a research assistant of the Fonds voor Wetenschappelijk Onderzoek-Vlaanderen. Frank Jacobs is a research assistant of the Instituut voor de Aanmoediging van Innovatie door Wetenschap en Technologie in Vlaanderen.

Author information

Author notes

  1. E Van Craeyveld and J Lievens: These authors contributed equally to this work.

Authors and Affiliations

  1. Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium

    E Van Craeyveld, J Lievens, F Jacobs, Y Feng, J Snoeys & B De Geest

Authors
  1. E Van Craeyveld
    View author publications

    Search author on:PubMed Google Scholar

  2. J Lievens
    View author publications

    Search author on:PubMed Google Scholar

  3. F Jacobs
    View author publications

    Search author on:PubMed Google Scholar

  4. Y Feng
    View author publications

    Search author on:PubMed Google Scholar

  5. J Snoeys
    View author publications

    Search author on:PubMed Google Scholar

  6. B De Geest
    View author publications

    Search author on:PubMed Google Scholar

Corresponding author

Correspondence to B De Geest.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Van Craeyveld, E., Lievens, J., Jacobs, F. et al. Apolipoprotein A-I and lecithin:cholesterol acyltransferase transfer induce cholesterol unloading in complex atherosclerotic lesions. Gene Ther 16, 757–765 (2009). https://doi.org/10.1038/gt.2009.8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue date:

  • DOI: https://doi.org/10.1038/gt.2009.8

Keywords

This article is cited by

Search

Advanced search

Quick links