gms | German Medical Science

24. Jahrestagung der Deutschen Gesellschaft für Arterioskleroseforschung

Deutsche Gesellschaft für Arterioskleroseforschung

18.03. - 20.03.2010, Blaubeuren

Role of GDF-15 (growth differentiation factor 15) in the progression of atherosclerosis in apoE deficient mice

Meeting Contribution

  • corresponding author S. Zügel - Anatomy and Cell Biology, Philipps-University Marburg, Germany
  • G. A. Bonaterra - Anatomy and Cell Biology, Philipps-University Marburg, Germany
  • S. Vorwald - Anatomy and Cell Biology III, Ruprecht-Karls-University Heidelberg, Germany
  • G. Bendner - Anatomy and Cell Biology III, Ruprecht-Karls-University Heidelberg, Germany
  • J. Strelau - Anatomy and Cell Biology III, Ruprecht-Karls-University Heidelberg, Germany
  • R. Kinscherf - Anatomy and Cell Biology, Philipps-University Marburg, Germany

Deutsche Gesellschaft für Arterioskleroseforschung e.V.. 24. Jahrestagung der Deutschen Gesellschaft für Arterioskleroseforschung. Blaubeuren, 18.-20.03.2010. Düsseldorf: German Medical Science GMS Publishing House; 2011. Doc10dgaf12

DOI: 10.3205/10dgaf12, URN: urn:nbn:de:0183-10dgaf120

Veröffentlicht: 23. März 2011

© 2011 Zügel et al.
Dieser Artikel ist ein Open Access-Artikel und steht unter den Creative Commons Lizenzbedingungen (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.de). Er darf vervielfältigt, verbreitet und öffentlich zugänglich gemacht werden, vorausgesetzt dass Autor und Quelle genannt werden.


Abstract

Growth differentiation factor-15 (GDF-15) is a member of the TGF-β superfamily as well as an important regulator of proliferation, differentiation and inflammatory processes and is found in macrophages of human atherosclerotic plaque. Hence the goal of the study was to investigate the effect of GDF-15 in the development and progression of atherosclerosis in apoE deficient (apoE-/-) mice.


Introduction

Growth differentiation factor-15 (GDF-15), also known as macrophage inhibitory cytokine-1 (MIC-1), is a member of the transforming growth factor (TGF-β) superfamily. TGF-βs are cytokines that exert prominent functions in tissue homeostasis and adaptation by regulating cell survival, proliferation, differentiation and inflammatory processes. It has been shown that GDF-15 is involved in the regulation of proapoptotic/antiinflammatory processes under contribution of p53 and poly-ADP-polymerase (PARP), interleukin-1 (IL-1), cyclooxygenase-1/2 (COX-1, COX-2) or tumor necrosis factor-α (TNF-α) [1], [2], [3], [4], [5], [6]. Additionally, oxidized low density lipoproteins (oxLDL), which are involved in the development of atherosclerosis, induce the expression of GDF-15 in human macrophages with a simultaneous increase of the apoptosis rate [7]. Moreover, it has recently been shown that GDF-15 and oxLDL are colocalized in human macrophages of atherosclerotic lesions [7]. Thus, the aim of our study was to investigate the effect of GDF-15 in the development and progression of atherosclerosis in apoE deficient (apoE-/-) mice.


Methods

ApoE-/-/GDF-15+/+ and apoE-/-/GDF-15-/- mice were generated by cross-breeding of apoE-/- and GDF-15 LacZ-knockin mice. At an age of 10 weeks, the offspring were fed with a high cholesterol diet (western diet) for a period of 12 weeks. Thereafter, the innominate artery (=brachiocephalic trunk) was removed, shock-frozen in liquid nitrogen-cooled isopentane and 6 µm cryo-sections were generated. The extent of atherosclerotic plaques was measured by computer-assisted morphometry (Figure 1 [Fig. 1]). Detection of BM8 and MOMA-2 (macrophages), Ki67 (proliferation), COX-2 and MIF (inflammation) was quantified via immunohistological staining including computer-assisted morphometry.


Results

In apoE-/-/GDF-15-/- mice we found an inhibition of lumen stenosis of 17% (p=0.05) compared to apoE-/-/GDF-15+/+ mice (Figure 2 [Fig. 2]). Furthermore, the density (cells/mm²) of COX-2 (+91%, p=0.02; Figure 3 [Fig. 3]) and MIF (+202%, p=0.04; Figure 4 [Fig. 4]) positive cells was increased in atherosclerotic lesions of apoE-/-/GDF-15-/- mice compared to apoE-/-/GDF-15+/+ mice. Additionally, in atherosclerotic lesions of apoE-/-/GDF-15-/- the density of Ki67 positive cells was significantly increased (+94%, p=0.05), whereas the percentage of BM8 and MOMA-2 positive cells was similar in atherosclerotic lesions of both groups under test (not shown).


Discussion

Previous data have shown that GDF-15 immunoreactivity is localized in macrophages of human atherosclerotic lesions [7]. Thus, it could be hypothesized that GDF-15 plays a role during development and progression of atherosclerosis. Indeed, we show here that GDF-15 deficiency significantly inhibits the progression of atherosclerotic plaques in the innominate artery of cholesterol-fed apoE-/- mice. Furthermore, atherosclerosis is an inflammatory disease [8] and proinflammatory as well as proapoptotic cytokines have been shown to be co-localized in cells of human atherosclerotic lesions [9], [10]. According to previous data in humans, we found in atherosclerotic plaques of apoE-/- mice MIF and COX-2 immunoreactive cells. Because GDF-15 deficiency increased the expression of MIF and COX-2, it is tempting to speculate that GDF-15 is involved in expression of these pro-inflammatory cytokines. Moreover, an increased density of MIF and COX-2 expressing cells seems to result in an enhanced pro-inflammatory status of the plaque area. MIF has been shown to be produced by macrophages in all types of atherosclerotic lesions, where it has been suggested to play a significant role by maintaining the activation-mediated induction of inflammatory processes [10], [11] and to play a role in the destabilization of human atherosclerotic plaque [12]. However, we found that lumen stenosis is reduced in apoE-/-/GDF-15-/- mice, which may indicate that GDF-15 is associated with plaque stability/instability status. In macrophages of human atherosclerotic lesions a co-localization of GDF-15 with oxLDL as well as with inflammatory and/or apoptotic markers were likewise found [7], so that it can be hypothesized that GDF-15 is an early mediator of damage, which could adjust inflammation [9] survival, proliferation or apoptosis during illness processes [13]. Finally, according to our results, we assume that GDF-15 supports the progression of atherosclerosis, however independent from induction of inflammatory reactions and proliferative processes.


References

1.
Fairlie WD, Moore AG, Bauskin AR, Russell PK, Zhang HP, Breit SN. MIC-1 is a novel TGF-beta superfamily cytokine associated with macrophage activation. J Leukoc Biol. 1999;65:2-5.
2.
Wilson LC, Baek SJ, Call A, Eling TE. Nonsteroidal anti-inflammatory drug-activated gene (NAG-1) is induced by genistein through the expression of p53 in colorectal cancer cells. Int J Cancer. 2003;105(6):747-53. DOI: 10.1002/ijc.11173 Externer Link
3.
Kim JS, Baek SJ, Sali T, Eling TE. The conventional nonsteroidal anti-inflammatory drug sulindac sulfide arrests ovarian cancer cell growth via the expression of NAG-1/MIC-1/GDF-15. Mol Cancer Ther. 2005;4(3):487-93.
4.
Kim KS, Yoon JH, Kim JK, Baek SJ, Eling TE, Lee WJ, Ryu JH, Lee JG, Lee JH, Yoo JB. Cyclooxygenase inhibitors induce apoptosis in oral cavity cancer cells by increased expression of nonsteroidal anti-inflammatory drug-activated gene. Biochem Biophys Res Commun. 2004;325(4):1298-303. DOI: 10.1016/j.bbrc.2004.10.176 Externer Link
5.
Baek SJ, Kim JS, Moore SM, Lee SH, Martinez J, Eling TE. Cyclooxygenase inhibitors induce the expression of the tumor suppressor gene EGR-1, which results in the up-regulation of NAG-1, an antitumorigenic protein. Mol Pharmacol. 2005; 67(2):356-64. DOI: 10.1124/mol.104.005108 Externer Link
6.
Kadara H, Schroeder CP, Lotan D, Pisano C, Lotan R. Induction of GDF-15/NAG-1/MIC-1 in human lung carcinoma cells by retinoid-related molecules and assessment of its role in apoptosis. Cancer Biol Ther. 2006; 5(5):518-22. DOI: 10.4161/cbt.5.5.2602 Externer Link
7.
Schlittenhardt D, Schober A, Strelau J, Bonaterra GA, Schmiedt W, Unsicker K, Metz J, Kinscherf R. Involvement of growth differentiation factor-15/ macrophage inhibitory cytokine-1 (GDF-15/MIC-1) in oxLDL-induced apoptosis of human macrophages in vitro and in arteriosclerotic lesions. Cell Tissue Res. 2004;318(2): 325-33. DOI: 10.1007/s00441-004-0986-3 Externer Link
8.
Ross R. Atherosclerosis-an inflammatory disease. N Engl J Med. 1999;340:115- 26. DOI: 10.1056/NEJM199901143400207 Externer Link
9.
Kinscherf R, Wagner M, Kamencic H, Bonaterra GA, Hou D, Schiele RA, Deigner HP, Metz J. Characterization of apoptotic macrophages in atheromatous tissue of humans and heritable hyperlipidemic rabbits. Atherosclerosis. 1999;144(1):33-9. DOI: 10.1016/S0021-9150(99)00037-4 Externer Link
10.
Burger-Kentischer A, Goebel H, Seiler R, Fraedrich G, Schaefer HE, Dimmeler S, Kleemann R, Bernhagen J, Ihling C. Expression of macrophage migration inhibitory factor in different stages of human atherosclerosis. Circulation. 2002;105:1561-6. DOI: 10.1161/01.CIR.0000012942.49244.82 Externer Link
11.
Zernecke A, Weber C. Inflammatory mediators in atherosclerotic vascular disease. Basic Res Cardiol. 2005;100:93-101. DOI: 10.1007/s00395-005-0511-6 Externer Link
12.
Kong YZ, Huang XR, Ouyang X, Tan JJ, Fingerle-Rowson G, Bacher M, Mu W, Scher LA, Leng L, Bucala R, Lan HY. Evidence for vascular macrophage migration inhibitory factor in destabilization of human atherosclerotic plaques. Cardiovasc Res. 2005;65:272-82. DOI: 10.1016/j.cardiores.2004.09.020 Externer Link
13.
Hansson G. Inflammation, atherosclerosis, and coronary artery disease. N Engl J Med. 2005;352:1685-95. DOI: 10.1056/NEJMra043430 Externer Link