Coexisting proinflammatory and antioxidative endothelial transcription profiles in a disturbed flow region of the adult porcine aorta.

In the arterial circulation, regions of disturbed flow (DF), which are characterized by flow separation and transient vortices, are susceptible to atherogenesis, whereas regions of undisturbed laminar flow (UF) appear protected. Coordinated regulation of gene expression by endothelial cells (EC) may result in differing regional phenotypes that either favor or inhibit atherogenesis. Linearly amplified RNA from freshly isolated EC of DF (inner aortic arch) and UF (descending thoracic aorta) regions of normal adult pigs was used to profile differential gene expression reflecting the steady state in vivo. By using human cDNA arrays, ≈2,000 putatively differentially expressed genes were identified through false-discovery-rate statistical methods. A sampling of these genes was validated by quantitative real-time PCR and/or immunostaining en face. Biological pathway analysis revealed that in DF there was up-regulation of several broad-acting inflammatory cytokines and receptors, in addition to elements of the NF-κB system, which is consistent with a proinflammatory phenotype. However, the NF-κB complex was predominantly cytoplasmic (inactive) in both regions, and no significant differences were observed in the expression of key adhesion molecules for inflammatory cells associated with early atherogenesis. Furthermore, there was no histological evidence of inflammation. Protective profiles were observed in DF regions, notably an enhanced antioxidative gene expression. This study provides a public database of regional EC gene expression in a normal animal, implicates hemodynamics as a contributory mechanism to athero-susceptibility, and reveals the coexistence of pro- and antiatherosclerotic transcript profiles in susceptible regions. The introduction of additional risk factors may shift this balance to favor lesion development.

[1]  C. Napoli,et al.  Beneficial effects of antioxidants and l-arginine on oxidation-sensitive gene expression and endothelial NO synthase activity at sites of disturbed shear stress , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[2]  Ruediger C. Braun-Dullaeus,et al.  Vascular proliferation and atherosclerosis: New perspectives and therapeutic strategies , 2002, Nature Medicine.

[3]  Elisabetta Manduchi,et al.  Fidelity and enhanced sensitivity of differential transcription profiles following linear amplification of nanogram amounts of endothelial mRNA. , 2003, Physiological genomics.

[4]  Richard T. Lee,et al.  Vitamin D3–Upregulated Protein-1 (VDUP-1) Regulates Redox-Dependent Vascular Smooth Muscle Cell Proliferation Through Interaction With Thioredoxin , 2002, Circulation research.

[5]  R. Nerem,et al.  Oscillatory and steady laminar shear stress differentially affect human endothelial redox state: role of a superoxide-producing NADH oxidase. , 1998, Circulation research.

[6]  Larry V. McIntire,et al.  DNA microarray reveals changes in gene expression of shear stressed human umbilical vein endothelial cells , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[7]  R. Tibshirani,et al.  Significance analysis of microarrays applied to the ionizing radiation response , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[8]  T. Collins,et al.  Nuclear factor-kappa B interacts functionally with the platelet-derived growth factor B-chain shear-stress response element in vascular endothelial cells exposed to fluid shear stress. , 1995, The Journal of clinical investigation.

[9]  E. R. Taylor,et al.  Isolation and characterization of two novel A20-like proteins , 2001 .

[10]  F. Spriggs,et al.  Gene expression profile of human endothelial cells exposed to sustained fluid shear stress. , 2002, Physiological genomics.

[11]  A. Pries,et al.  Evidence for modulation of genes involved in vascular adaptation by prolonged exposure of endothelial cells to shear stress. , 2000, Cardiovascular research.

[12]  S. Usami,et al.  Molecular mechanism of endothelial growth arrest by laminar shear stress. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[13]  C. J. Gimeno,et al.  Vascular MADs: two novel MAD-related genes selectively inducible by flow in human vascular endothelium. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[14]  R. Ross,et al.  Studies of Hypercholesterolemia in the Nonhuman Primate: I. Changes that Lead to Fatty Streak Formation , 1984, Arteriosclerosis.

[15]  D. Sorescu,et al.  NAD(P)H oxidase: role in cardiovascular biology and disease. , 2000, Circulation research.

[16]  Andrew C. Li,et al.  The macrophage foam cell as a target for therapeutic intervention , 2002, Nature Medicine.

[17]  R. Knuechel,et al.  Activated transcription factor nuclear factor-kappa B is present in the atherosclerotic lesion. , 1996, The Journal of clinical investigation.

[18]  G. Garcı́a-Cardeña,et al.  Biomechanical activation of vascular endothelium as a determinant of its functional phenotype , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[19]  M. Cybulsky,et al.  NF-κB: pivotal mediator or innocent bystander in atherogenesis? , 2001 .

[20]  Amy Milsted,et al.  Shear stress magnitude and directionality modulate growth factor gene expression in preconditioned vascular endothelial cells. , 2003, Journal of vascular surgery.

[21]  K. O. Mercurius,et al.  Stimulation of transcription factors NF kappa B and AP1 in endothelial cells subjected to shear stress. , 1994, Biochemical and biophysical research communications.

[22]  P. Lelkes,et al.  Gene expression profiling of human aortic endothelial cells exposed to disturbed flow and steady laminar flow. , 2002, Physiological genomics.

[23]  B. Chen,et al.  DNA microarray analysis of gene expression in endothelial cells in response to 24-h shear stress. , 2001, Physiological genomics.

[24]  P. Libby,et al.  Stabilization of atherosclerotic plaques: New mechanisms and clinical targets , 2002, Nature Medicine.

[25]  J. Cornhill,et al.  A quantitative study of the localization of atherosclerotic lesions in the rabbit aorta. , 1976, Atherosclerosis.

[26]  Zaverio M. Ruggeri,et al.  Platelets in atherothrombosis , 2002, Nature Medicine.

[27]  M. Cybulsky,et al.  Patterns of vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 expression in rabbit and mouse atherosclerotic lesions and at sites predisposed to lesion formation. , 1999, Circulation research.

[28]  M. Cybulsky,et al.  The NF-kappa B signal transduction pathway in aortic endothelial cells is primed for activation in regions predisposed to atherosclerotic lesion formation. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[29]  R. Nadon,et al.  Statistical issues with microarrays: processing and analysis. , 2002, Trends in genetics : TIG.

[30]  D. Rader,et al.  The adhesion receptor CD44 promotes atherosclerosis by mediating inflammatory cell recruitment and vascular cell activation. , 2001, The Journal of clinical investigation.

[31]  J. Witztum,et al.  Innate and acquired immunity in atherogenesis , 2002, Nature Medicine.

[32]  M. Reidy,et al.  Scanning electron microscopy in the evaluation of endothelial integrity of the fatty lesion in atherosclerosis. , 1976, Atherosclerosis.

[33]  M. Gimbrone,et al.  Identification of vascular endothelial genes differentially responsive to fluid mechanical stimuli: cyclooxygenase-2, manganese superoxide dismutase, and endothelial cell nitric oxide synthase are selectively up-regulated by steady laminar shear stress. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[34]  D. Wagner,et al.  Localized reduction of atherosclerosis in von Willebrand factor-deficient mice. , 2001, Blood.

[35]  P. Benos,et al.  Genomic analysis of immediate/early response to shear stress in human coronary artery endothelial cells. , 2002, Physiological genomics.

[36]  M. Gimbrone,et al.  The Critical Role of Mechanical Forces in Blood Vessel Development, Physiology and Pathology , 1999 .

[37]  F. Hosoda,et al.  A BAC-based STS-content map spanning a 35-Mb region of human chromosome 1p35-p36. , 2001, Genomics.

[38]  Daniel Steinberg,et al.  Atherogenesis in perspective: Hypercholesterolemia and inflammation as partners in crime , 2002, Nature Medicine.

[39]  Guang-Zhong Yang,et al.  Helical and Retrograde Secondary Flow Patterns in the Aortic Arch Studied by Three‐Directional Magnetic Resonance Velocity Mapping , 1993, Circulation.

[40]  D. Ku,et al.  Hemodynamics and atherosclerosis. Insights and perspectives gained from studies of human arteries. , 1988, Archives of pathology & laboratory medicine.

[41]  D. Mangelsdorf,et al.  The liver X receptor gene team: Potential new players in atherosclerosis , 2002, Nature Medicine.

[42]  N Harbeck,et al.  Spatial and temporal regulation of gap junction connexin43 in vascular endothelial cells exposed to controlled disturbed flows in vitro. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[43]  Michael Karin,et al.  Is NF‐κB the sensor of oxidative stress? , 1999 .

[44]  P. Baeuerle,et al.  IKAP is a scaffold protein of the IκB kinase complex , 1998, Nature.

[45]  M. Gimbrone,et al.  Shear stress selectively upregulates intercellular adhesion molecule-1 expression in cultured human vascular endothelial cells. , 1994, The Journal of clinical investigation.

[46]  C. Kunsch,et al.  Laminar Flow Induction of Antioxidant Response Element-mediated Genes in Endothelial Cells , 2003, The Journal of Biological Chemistry.

[47]  J. Eberwine,et al.  Amplified RNA synthesized from limited quantities of heterogeneous cDNA. , 1990, Proceedings of the National Academy of Sciences of the United States of America.