Analysis of gene and protein expression during monocyte-macrophage differentiation and cholesterol loading--cDNA and protein array study.

BACKGROUND: To better understand the role of macrophages in atherogenesis and to find new strategies to prevent their harmful effects, more information is needed about their gene and protein expression patterns in atherogenic conditions. METHODS: We analyzed gene and protein expression changes during monocyte-macrophage differentiation and lipid-loading by cDNA arrays and antibody-based protein arrays, respectively. RESULTS: It was found that early response genes, such as transcription factors, were upregulated early during monocyte-macrophage differentiation, while genes functioning in cell proliferation, migration, inflammation and lipid metabolism were activated later during macrophage differentiation. When comparing results from cDNA and antibody arrays, it become evident that changes at the protein levels were not always predicted by changes at the mRNA level. This discrepancy may be due to the different transcript variants that exist for distinct genes, posttranslational modifications and different turnover rates for mRNAs and proteins of distinct genes. When combining cDNA and protein array results with RT-PCR, it was found that CD36, COX-2, and several factors regulating cell signaling, such as Cdk-1, TFII-I, NEMO-like kinase, Elf-5 and TRADD were strongly upregulated both at the mRNA and protein levels. CONCLUSIONS: Time-dependency of the activation of the early response genes and genes functioning in inflammation, lipid metabolism and cell proliferation and migration, is an important feature of the macrophage differentiation. It was also evident that several novel transcription factors were activated during lipid-loading. It is concluded that cDNA and protein arrays will be useful for the identification of genes that are potential targets for therapeutic interventions.

Gene expression in macrophage-rich inflammatory cell infiltrates in human atherosclerotic lesions as studied by laser microdissection and DNA array: overexpression of HMG-CoA reductase, colony stimulating factor receptors, CD11A/CD18 integrins, and interleukin receptors.

OBJECTIVE: Inflammatory cells play an important role in atherogenesis. However, more information is needed about their gene expression profiles in human lesions. METHODS AND RESULTS: We used laser microdissection (LMD) to isolate macrophage-rich shoulder areas from human lesions. Gene expression profiles in isolated cells were analyzed by cDNA array and compared with expression patterns in normal intima and THP-1 macrophages. Upregulation of 72 genes was detected with LMD and included 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, interferon regulatory factor-5 (IRF-5), colony stimulating factor (CSF) receptors, CD11a/CD18 integrins, interleukin receptors, CD43, calmodulin, nitric oxide synthase (NOS), and extracellular superoxide dismutase (SOD). Several of these changes were also present in PMA-stimulated THP-1 macrophages in vitro. On the other hand, expression of several genes, such as VEGF, tissue factor pathway inhibitor 2, and apolipoproteins C-I and C-II, decreased. CONCLUSIONS: Overexpression of HMG-CoA reductase in macrophage-rich lesion areas may explain some beneficial effects of statins, which can also modulate increased expression of CD11a/CD18 and CD43 found in microdissected cells. We also found increased expression of CSF receptors, IRF-5, and interleukin receptors, which could become useful therapeutic targets for the treatment of atherosclerotic diseases.