Functional genomics and DNA array techniques in atherosclerosis research

Mo, Hiltunen; Niemi, Mari; Ylä‐Herttuala, Seppo

doi:10.1097/00041433-199912000-00006

Cited by 28 publications

(11 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, expression of these genes in vivo remains to be determined. Other groups focused on differences in gene expression between fatty streaks and advanced lesions 10 and intima and media of human atherosclerotic plaques. 11 Adams et al 12 revealed different gene expression patterns between veins and arteries in macaques.…”

mentioning

confidence: 99%

Identification of Genes Potentially Involved in Rupture of Human Atherosclerotic Plaques

Faber

Cleutjens

Niessen

et al. 2001

Circulation Research

126

View full text Add to dashboard Cite

Abstract-Although rupture of an atherosclerotic plaque is the major cause of acute vascular occlusion, the exact molecular mechanisms underlying this process are still poorly understood. In this study, we used suppression subtractive hybridization to make an inventory of genes that are differentially expressed in whole-mount human stable and ruptured plaques. Two libraries were generated, one containing 3000 clones upregulated and one containing 2000 clones downregulated in ruptured plaques. Macroarray analysis of 500 randomly chosen clones showed differential expression of 45 clones. Among the 25 clones that showed at least a 2-fold difference in expression was the gene of perilipin, upregulated in ruptured plaques, and the genes coding for fibronectin and immunoglobulin chain, which were downregulated in ruptured plaques. Reverse transcriptase-polymerase chain reaction analysis on 10 individual ruptured and 10 individual stable plaques showed a striking consistency of expression for the clones SSH6, present in 8 ruptured and 2 stable plaques, and perilipin, expressed in 8 ruptured plaques and completely absent in stable plaques.

show abstract

mentioning

confidence: 99%

Identification of Genes Potentially Involved in Rupture of Human Atherosclerotic Plaques

Faber

Cleutjens

Niessen

et al. 2001

Circulation Research

126

View full text Add to dashboard Cite

show abstract

“…We found 17 apoptosis-related genes, including 2 housekeeping genes, with a 2-to 5-fold difference in expression level between carotid endarterectomy specimens and nonatherosclerotic mammary arteries. Although gene expression profiling using cDNA array techniques seems to be an elegant way to identify novel genes or pathways that may contribute to features of atherosclerosis, 16 some pitfalls and/or study limitations should be considered when using this type of technology for the identification of apoptosis-related genes in atherosclerosis. First, apoptosis is a major event in the pathophysiology of atherosclerosis in which many proteins are involved.…”

Section: Discussionmentioning

confidence: 99%

Gene Expression Profiling of Apoptosis-Related Genes in Human Atherosclerosis

Martinet

Schrijvers

Meyer

et al. 2002

ATVB

View full text Add to dashboard Cite

Objective-Apoptosis substantially affects the cellularity and integrity of atherosclerotic plaques. It remains, however, unclear which key regulatory genes are involved. In this study, cDNA expression arrays were used to analyze transcript levels of 205 apoptosis-related genes in human carotid endarterectomy specimens versus nonatherosclerotic mammary arteries. Methods and Results-Seventeen genes with a 2-to 5-fold relative expression difference were identified. One of the most apparent changes in human plaques was the overexpression of death-associated protein (DAP) kinase (Ϸ5-fold), a positive mediator of apoptotic cell death. Differential expression of DAP kinase mRNA in human plaques relative to mammary arteries was confirmed by quantitative reverse-transcription polymerase chain reaction. Western blotting and immunohistochemistry demonstrated enhanced levels of DAP kinase protein in the plaque with negligible expression in non-atherosclerotic vessels. DAP kinase was located predominantly in foam cells of smooth muscle cell (SMC) origin. Uptake of aggregated LDL by cultured aortic SMCs as well as exposure of SMCs to the short-chain acyl ceramide derivative N-hexanoyl-D-sphingosine (C 6 -ceramide) upregulated DAP kinase both at the mRNA and protein level. Conclusions-Our data demonstrate that cDNA array technology can identify novel genes that might participate in cell death pathways underlying atherogenesis. Key Words: atherosclerosis Ⅲ apoptosis Ⅲ cDNA array Ⅲ death-associated protein kinase Ⅲ gene expression A poptosis has been identified as a prominent feature of advanced human atherosclerotic plaques. [1][2][3] The distribution of apoptosis within the plaque as determined by in situ end-labeling techniques (terminal deoxynucleotidyl transferase end labeling [TUNEL] and in situ nick translation [ISNT]) is heterogeneous, being more frequent (1% to 2%) in regions with a high density of macrophages. Early lesions that mainly consist of smooth muscle cells (SMCs) present very little apoptosis (Ͻ0.1%), suggesting that apoptosis in atherosclerotic plaques is strongly related to macrophage infiltration. In recent years, several factors with proapoptotic activity have been identified in human atherosclerotic plaques. SMCs within human fatty streaks or advanced plaques show increased expression of the proapoptotic protein Bax. 3 Furthermore, cytokines with proapoptotic potential, such as interleukin-1, tumor necrosis factor (TNF)-␣, and interferon (IFN)-␥ are secreted by inflammatory cells and contribute to local cell death. 1 Together with macrophages and T lymphocytes, mast cells are present in the inflammatory infiltrate and initiate SMCs apoptosis by releasing chymase in the extracellular matrix. 4 Upregulation of matrix metalloproteinases in human plaques may result in cell detachment from the matrix and apoptotic cell death (anoikis). Macrophages can also induce SMCs apoptosis via Fas/Fas-L interactions. 5 Other substances that are known to induce apoptosis in the plaque include oxidized LDLs, oxysterols, a...

show abstract

“…[1][2][3][4][5][6][7][8][9] In these studies a variety of techniques has been applied including cDNA expression arrays, differential display, RNA subtraction methods, serial analysis of gene expression (SAGE), and signature pyrosequencing, which have been performed both on whole-mount material and cell lines reported to be involved in atherosclerosis. Most of these studies focused on genes known to be involved in processes related to atherosclerosis such as inflammation, 10 apoptosis, 11 matrix turnover, 12 and lipid metabolism 13 or on well-known genes whose relationship to atherosclerosis had not been reported before.…”

Section: Introductionmentioning

confidence: 99%