Gene profiling in atherosclerosis reveals a key role for small inducible cytokines : Validation using a novel monocyte chemoattractant protein monoclonal antibody

• Published in: Circulation (New York, N.Y.) Vol. 111; no. 25; pp. 3443 - 3452
• Main Authors: LUTGENS, Esther, FABER, Birgit, MACKAY, Charles R, RENNERT, Paul, BAILLY, Veronique, JARPE, Matthew, DOLINSKI, Brian, KOTELIANSKY, Victor, DE FOUGEROLLES, Tony, DAEMEN, Mat J. A. P, SCHAPIRA, Kitty, EVELO, Chris T. A, VAN HAAFTEN, Rachel, HEENEMAN, Sylvia, CLEUTJENS, Kitty B. J. M, BIJNENS, Ann Pascale, BECKERS, Linda, PORTER, J. Gordon
• Format: Journal Article
• Language: English
• Published: Hagerstown, MD Lippincott Williams & Wilkins 28.06.2005
• Subjects: Animals; Antibodies, Monoclonal - administration & dosage; Antibodies, Monoclonal - pharmacology; Aorta, Thoracic; Apolipoproteins E - deficiency; Associated diseases and complications; Atherosclerosis (general aspects, experimental research); Atherosclerosis - drug therapy; Atherosclerosis - genetics; Atherosclerosis - pathology; Biological and medical sciences; Blood and lymphatic vessels; Cardiology. Vascular system; Chemokine CCL2 - immunology; Chemokine CCL8; Chemokines - genetics; Chemokines - physiology; Cluster Analysis; Diabetes. Impaired glucose tolerance; Disease Progression; Diseases of the peripheral vessels. Diseases of the vena cava. Miscellaneous; Endocrine pancreas. Apud cells (diseases); Endocrinopathies; Extracellular Matrix - metabolism; Gene Expression Profiling; Inflammation - genetics; Male; Medical sciences; Mice; Mice, Knockout; Monocyte Chemoattractant Proteins - immunology; Monocyte Chemoattractant Proteins - physiology; Peptide Hydrolases - genetics; RNA, Messenger - analysis; Time Factors; Vascular disease; Monocyte; genes; Atherosclerosis; Cytokine; Cardiovascular disease; Inflammation; Monoclonal antibody
• ISSN: 0009-7322; 1524-4539
• DOI: 10.1161/CIRCULATIONAHA.104.510073

Pathological aspects of atherosclerosis are well described, but gene profiles during atherosclerotic plaque progression are largely unidentified. Microarray analysis was performed on mRNA of aortic arches of ApoE-/- mice fed normal chow (NC group) or Western-type diet (WD group) for 3, 4.5, and 6 months. Of 10 176 reporters, 387 were differentially (>2x) expressed in at least 1 group compared with a common reference (ApoE-/-, 3- month NC group). The number of differentially expressed genes increased during plaque progression. Time-related expression clustering and functional grouping of differentially expressed genes suggested important functions for genes involved in inflammation (especially the small inducible cytokines monocyte chemoattractant protein [MCP]-1, MCP-5, macrophage inflammatory protein [MIP]-1alpha, MIP-1beta, MIP-2, and fractalkine) and matrix degradation (cathepsin-S, matrix metalloproteinase-2/12). Validation experiments focused on the gene cluster of small inducible cytokines. Real-time polymerase chain reaction revealed a plaque progression-dependent increase in mRNA levels of MCP-1, MCP-5, MIP-1alpha, and MIP-1beta. ELISA for MCP-1 and MCP-5 showed similar results. Immunohistochemistry for MCP-1, MCP-5, and MIP-1alpha located their expression to plaque macrophages. An inhibiting antibody for MCP-1 and MCP-5 (11K2) was designed and administered to ApoE-/- mice for 12 weeks starting at the age of 5 or 17 weeks. 11K2 treatment reduced plaque area and macrophage and CD45+ cell content and increased collagen content, thereby inducing a stable plaque phenotype. Gene profiling of atherosclerotic plaque progression in ApoE-/- mice revealed upregulation of the gene cluster of small inducible cytokines. Further expression and in vivo validation studies showed that this gene cluster mediates plaque progression and stability.