In Vitro-In Vivo Gene Expression Analysis in Atherosclerosis
Atherosclerosis, the pathologic inflammatory response to injury. of the human vessel wall, has been long recognized for its complexity of initiation, progression and ultimate appearance of clinical symptoms . Many proteins and other compounds have been implicated in atherogenesis, and this list is now growing exponentially with the recent advances in high-throughput gene expression profiling [1 2 3 4 5 6 7 8 9].Indeed, a plethora of individual genes show altered expression during atherosclerosis, but the development of intervention strategies based on such individual genes in animal models has been rather challenging. The translation into treatment of atherosclerosis in man has proven even more difficult. A clear gene-environment interaction, most notably Western-type diet and life-style, lies at the basis of disease development. This indicates that disturbed patterns of gene-expression rather than single culprit genes form the basis for the widespread penetrance of the disease in the elderly Western population. We are applying functional Genomics to the study of atherosclerosis, with the goal of characterizing healthy and diseased gene expression profiles. While our immediate objective is to characterize those genes that are differentially expressed during atherogenesis, our long-term goal is to determine how a healthy gene expression profile can be induced in the cells of the vascular wall. This implies not only to identify differentially expressed genes but also to determine their function and, most importantly, to analyze the integrated pathways and mechanisms through which their expression is regulated. In this report we will describe the use of differential display RT-PCR and cDNA microarray expression analysis to determine changes in gene expression profiles in cultured vascular endothelial cells in response to pro-and antiatherogenic stimuli. We will briefly explore the computational analysis of such gene expression profiles as detected by a custom cardiovascular microarray. Finally, we show that insights that were gained in vitro can be extended to the in vivo (atherosclerotic) vascular wall.
KeywordsCholesterol Foam Prostaglandin Thrombin Triglyceride
Unable to display preview. Download preview PDF.
- 2.Horrevoets AJG, Fontijn RD, van Zonneveld A, de Vries CJM, ten Cate JW, and Pannekoek H. Vascular endothelial genes that are responsive to tumor necrosis factor-alpha in vitro are expressed in atherosclerotic lesions, including Inhibitor of Apoptosis Protein-1, stannin and two novel genes. BLOOD 1999;93:3418–31.PubMedGoogle Scholar
- 3.de Vries CJM, van Achterberg TAE, Horrevoets AJG, ten Cate JW, and Pannekoek H. Differential display identification of 40 genes with altered expression in activated human smooth muscle cells: local expression in atherosclerotic lesions of smags, smooth muscle activation-specific genes. J. Biol. Chem. 2000;275:23939–47.PubMedCrossRefGoogle Scholar
- 7.Shiffman D, Mikita T, Tai JT, Wade DP, Porter JG, Seilhamer JJ, Somogyi R, Liang S, Lawn RM.Google Scholar
- 8.Large-scale gene expression analysis of cholesterol-loaded macrophages. J Biol Chem. 2000;275:37324–32.Google Scholar
- 12.Monajemi H, Fontijn RD, Pannekoek H, and Horrevoets AJG. The Apolipoprotein L gene cluster has emerged recently in evolution and is expressed in human vascular tissue. Submitted.Google Scholar
- 13.Asakura T, Karino, T. Flow patterns and spatial distribution of atherosclerotic lesions in human coronary arteries Circ Res 1990;66:1045–66.Google Scholar
- 19.Dekker RJ, van Soest S, Pannekoek H, and Horrevoets AJG. A micro-array analysis of fluid shear-stress modulated genes in endothelial cells discriminates NFkB-dependent and independent pathways. Submitted.Google Scholar
- 20.Dekker RJ, Pannekoek H, and Horrevoets AJG. Lung Kruppel-like Factor is specifically upregulated by fluid shear-stress in endothelial cells in vitro and absent from atherosclerotic lesions in vivo. Submitted.Google Scholar