EICOSANOID SYNTHESIS AND FUNCTION IN CORONARY VESSELS

Summary

Principal Investigator: WILLIAM BRYSON CAMPBELL
Abstract: Endothelial cells (ECs) regulate vascular tone, leukocyte function and platelet aggregation through the release of several soluble mediators including PGI2, nitric oxide and endothelium-dependent hyperpolarizing factor (EDHF). In coronary arteries, hormones such as acetylcholine, bradykinin and arachidonic acid (AA) and physiological stimuli such as flow-induced shear stress produce endothelium-dependent vasodilation that is inhibited by cytochrome P450 (CYP) inhibitors. Thus, CYP metabolite(s) of AA contribute to agonist- induced relaxations. Coronary ECs synthesize vasodilator eicosanoids: PGI2 by cyclooxygenase and 14,15-, 11,12-, 8,9- and 5,6-epoxyeicosatrienoic acids (EETs) by CYP. EETs relax coronary arteries by opening calcium-activated potassium (KCa) channels and hyperpolarizing the smooth muscle. The proposed studies will test the hypothesis that endothelial metabolites of AA are involved in the regulation of coronary vascular tone. These studies will focus on the endothelium-derived EETs. We will test the hypothesis that EETs are EDHFs in the coronary vasculature by investigating the following specific aims: (1) We will identify and characterize analogs for the four EET regioisomers that act as selective EET agonists and antagonists. Regioisomer-specific antagonists will be developed. Stable agonists and antagonists that resist metabolism by vascular cells will be developed for use in vitro and importantly in vivo. These analogs will be used to define the role of EET isomers in regulating vascular tone and mediating dilation to agonists. (2) Vascular receptors for the EETs have not been characterized despite data suggesting their existence. We will determine if the EETs have specific binding sites/receptors on membranes from coronary arteries, ECs and smooth muscle cells using agonist analogs that incorporate iodine-125, biotin and photoaffinity groups and an agonist tethered to silica beads. 125I-14,15-EET analogs will be used as radioligands to characterize the receptor/binding site. Kinetics and ligand specificity of the receptor will be determined. The properties of EET binding proteins will be studied. (3) In response to agonists, ECs release mainly 14,15-EET with lesser amounts of 11,12- and 8,9-EET. EETs act stereospecifically in causing vascular relaxation. Thus, the total concentration of a released EET regioisomer may not reflect the concentration of the biologically active EET stereoisomer. The concentration and activity of the stereoisomers must be taken into account. We will determine the activity of the stereoisomers of the EET and DHET regioisomers. We will identify the stereoisomers of the EET regioisomers that are synthesized by ECs and coronary arteries and study the regulation of their release by agonists. These studies should indicate the major biologically active EET(s) released by coronary arteries. These studies will provide specific pharmacological tools to study the role of EETs as EDHFs and provide new insights into the vascular action of the EETs.
Funding Period: ----------------1994 - ---------------2011-
more information: NIH RePORT

Top Publications

  1. ncbi Regulation of potassium channels in coronary smooth muscle by adenoviral expression of cytochrome P-450 epoxygenase
    William B Campbell
    Dept of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Rd, Milwaukee, WI 53226, USA
    Am J Physiol Heart Circ Physiol 290:H64-71. 2006
  2. pmc Ecto-5'-nucleotidase, CD73, is an endothelium-derived hyperpolarizing factor synthase
    Masanori Ohta
    Department of Medicine and Cardiovascular Center, Medical College of Wisconsin, Milwaukee, Wisconsin, USA
    Arterioscler Thromb Vasc Biol 33:629-36. 2013
  3. pmc ACE inhibition enhances bradykinin relaxations through nitric oxide and B1 receptor activation in bovine coronary arteries
    Kathryn M Gauthier
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, WI, USA
    Biol Chem 394:1205-12. 2013
  4. pmc Effects of the selective EET antagonist, 14,15-EEZE, on cardioprotection produced by exogenous or endogenous EETs in the canine heart
    Garrett J Gross
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Rd, Milwaukee, WI 53226, USA
    Am J Physiol Heart Circ Physiol 294:H2838-44. 2008
  5. ncbi Identification and characterization of the unique guanine nucleotide exchange factor, SmgGDS, in vascular smooth muscle cells
    Rebecca Thill
    Department of Pharmacology and Toxicology, Cardiovascular Center, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA
    J Cell Biochem 104:1760-70. 2008
  6. ncbi Characterization of epoxyeicosatrienoic acid binding site in U937 membranes using a novel radiolabeled agonist, 20-125i-14,15-epoxyeicosa-8(Z)-enoic acid
    Wenqi Yang
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Rd, Milwaukee, WI 53226, USA
    J Pharmacol Exp Ther 324:1019-27. 2008
  7. ncbi Roles of epoxyeicosatrienoic acids in vascular regulation and cardiac preconditioning
    Kathryn M Gauthier
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, WI 53226, USA
    J Cardiovasc Pharmacol 50:601-8. 2007
  8. pmc Hydrogen peroxide inhibits cytochrome p450 epoxygenases: interaction between two endothelium-derived hyperpolarizing factors
    Brandon T Larsen
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Rd, Milwaukee, WI 53226, USA
    Circ Res 102:59-67. 2008
  9. ncbi Characterization of 14,15-epoxyeicosatrienoyl-sulfonamides as 14,15-epoxyeicosatrienoic acid agonists: use for studies of metabolism and ligand binding
    Wenqi Yang
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, USA
    J Pharmacol Exp Ther 321:1023-31. 2007
  10. pmc Epoxygenase eicosanoids: synthesis of tetrahydrofuran-diol metabolites and their vasoactivity
    J R Falck
    Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA
    Bioorg Med Chem Lett 17:2634-8. 2007

Scientific Experts

  • Garrett Gross
  • J Russell Falck
  • Kathryn M Gauthier
  • Kasem Nithipatikom
  • WILLIAM BRYSON CAMPBELL
  • Brandon T Larsen
  • Ishfaq A Bukhari
  • Yuenmu Chen
  • David D Gutterman
  • Wenqi Yang
  • Hiroto Miura
  • Fan Yi
  • Xiu Yu Yi
  • Masanori Ohta
  • Zhuohui Gan
  • Kazuyoshi Toyama
  • Sandra L Pfister
  • Bhavani Sangras
  • Rebecca Thill
  • Pin Lan Li
  • Darryl C Zeldin
  • Andrew Y Zhang
  • Iain N McSherry
  • Ryutaro Teraoka
  • Vincent Lemaitre
  • John D Imig
  • Abdul Jabbar Shah
  • Sreenivasulu Reddy Koduru
  • Katherine A Walsh
  • Vijaya L Manthati
  • Jawahar Lal Jat
  • Said H Audi
  • Robert D Bongard
  • Marilyn P Merker
  • Setti G Jagadeesh
  • Venugopal R Tuniki
  • Si Jin
  • Vijay L Manthati
  • Atsushi Sato
  • Carol L Williams
  • Siddam Anjaiah
  • Cecilia J Hillard
  • Venugopal Raju Tuniki
  • Blythe B Holmes
  • R V Krishna Kishore
  • Lijie Cui
  • V Raj Gopal
  • Kim A Dora
  • Bruce D Hammock
  • Ossama A Hatoum
  • Guo Zhang
  • Erich Gulbins
  • Shaun L Sandow
  • Michael A Hill

Detail Information

Publications28

  1. ncbi Regulation of potassium channels in coronary smooth muscle by adenoviral expression of cytochrome P-450 epoxygenase
    William B Campbell
    Dept of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Rd, Milwaukee, WI 53226, USA
    Am J Physiol Heart Circ Physiol 290:H64-71. 2006
    ..These studies indicate that 14(S),15(R)-EET is a sufficient endogenous activator of BK(Ca) channels in coronary SMCs...
  2. pmc Ecto-5'-nucleotidase, CD73, is an endothelium-derived hyperpolarizing factor synthase
    Masanori Ohta
    Department of Medicine and Cardiovascular Center, Medical College of Wisconsin, Milwaukee, Wisconsin, USA
    Arterioscler Thromb Vasc Biol 33:629-36. 2013
    ..We tested the hypothesis that coronary vasodilation to adenine nucleotides is mediated by an endothelial CD73-dependent, extracellular production of adenosine that acts as an endothelium-derived hyperpolarizing factor...
  3. pmc ACE inhibition enhances bradykinin relaxations through nitric oxide and B1 receptor activation in bovine coronary arteries
    Kathryn M Gauthier
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, WI, USA
    Biol Chem 394:1205-12. 2013
    ..Our results demonstrate that ACE inhibitor-enhanced bradykinin relaxations of bovine coronary arteries occur through endothelial cell B1 receptor activation and NO...
  4. pmc Effects of the selective EET antagonist, 14,15-EEZE, on cardioprotection produced by exogenous or endogenous EETs in the canine heart
    Garrett J Gross
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Rd, Milwaukee, WI 53226, USA
    Am J Physiol Heart Circ Physiol 294:H2838-44. 2008
    ..These data also suggest that 14,15-EEZE is not blocking the mitochondrial ATP-sensitive potassium channel as a mechanism for antagonizing the cardioprotective effects of the EETs...
  5. ncbi Identification and characterization of the unique guanine nucleotide exchange factor, SmgGDS, in vascular smooth muscle cells
    Rebecca Thill
    Department of Pharmacology and Toxicology, Cardiovascular Center, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA
    J Cell Biochem 104:1760-70. 2008
    ..Taken together these results identify SmgGDS as a novel regulator of myosin organization and contraction in VSMC...
  6. ncbi Characterization of epoxyeicosatrienoic acid binding site in U937 membranes using a novel radiolabeled agonist, 20-125i-14,15-epoxyeicosa-8(Z)-enoic acid
    Wenqi Yang
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Rd, Milwaukee, WI 53226, USA
    J Pharmacol Exp Ther 324:1019-27. 2008
    ..Using this radioligand, we have identified a specific high-affinity and high-abundance EET binding site in U937 cell membranes. This binding site could represent a specific EET receptor, which is probably a G protein-coupled receptor...
  7. ncbi Roles of epoxyeicosatrienoic acids in vascular regulation and cardiac preconditioning
    Kathryn M Gauthier
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, WI 53226, USA
    J Cardiovasc Pharmacol 50:601-8. 2007
    ..Production and metabolism of EETs as well as their specific cellular signaling mechanisms are discussed...
  8. pmc Hydrogen peroxide inhibits cytochrome p450 epoxygenases: interaction between two endothelium-derived hyperpolarizing factors
    Brandon T Larsen
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Rd, Milwaukee, WI 53226, USA
    Circ Res 102:59-67. 2008
    ..These findings suggest that an inhibitory interaction exists between 2 EDHFs in the human coronary microcirculation. CYP epoxygenases are directly inhibited by H2O2, and this interaction may modulate vascular EET bioavailability...
  9. ncbi Characterization of 14,15-epoxyeicosatrienoyl-sulfonamides as 14,15-epoxyeicosatrienoic acid agonists: use for studies of metabolism and ligand binding
    Wenqi Yang
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, USA
    J Pharmacol Exp Ther 321:1023-31. 2007
    ..14,15-EET-P(125)ISA is a new radiolabeled tool to study EET metabolism and binding. Our results also provide preliminary evidence that EETs exert their biological effect through a membrane binding site/receptor...
  10. pmc Epoxygenase eicosanoids: synthesis of tetrahydrofuran-diol metabolites and their vasoactivity
    J R Falck
    Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA
    Bioorg Med Chem Lett 17:2634-8. 2007
    ..The THFDs potently induced relaxation of pre-contracted bovine arteries...
  11. pmc Epoxyeicosatrienoic and dihydroxyeicosatrienoic acids dilate human coronary arterioles via BK(Ca) channels: implications for soluble epoxide hydrolase inhibition
    Brandon T Larsen
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, WI 53226, USA
    Am J Physiol Heart Circ Physiol 290:H491-9. 2006
    ..CYP450s and sEH may be endogenous sources of these compounds, and sEH inhibition has the potential to alter myocardial perfusion, depending on which EETs are produced endogenously...
  12. ncbi Lipid raft clustering and redox signaling platform formation in coronary arterial endothelial cells
    Andrew Y Zhang
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, USA
    Hypertension 47:74-80. 2006
    ..These results suggest that LR clustering occurs in coronary endothelial cells. The formation of redox signaling platforms on the cell membrane mediates transmembrane signaling of death receptors, resulting in endothelial dysfunction...
  13. ncbi A role for heterocellular coupling and EETs in dilation of rat cremaster arteries
    Iain N McSherry
    University of Bath, Bath, United Kingdom
    Microcirculation 13:119-30. 2006
    ..The authors probed endothelium-dependent dilation and endothelial cell Ca2+ handling in myogenically active resistance arteries...
  14. ncbi Beyond vasodilatation: non-vasomotor roles of epoxyeicosatrienoic acids in the cardiovascular system
    Brandon T Larsen
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, WI 53226, USA
    Trends Pharmacol Sci 28:32-8. 2007
    ..Finally, although usually protective in the systemic circulation, EETs might adversely affect the pulmonary circulation...
  15. ncbi Arachidonic acid metabolites as endothelium-derived hyperpolarizing factors
    William B Campbell
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, WI 53226, USA
    Hypertension 49:590-6. 2007
    ..These findings indicate that epoxyeicosatrienoic acids act as endothelium-derived hyperpolarizing factors and regulate arterial tone...
  16. ncbi Metabolism of adrenic acid to vasodilatory 1alpha,1beta-dihomo-epoxyeicosatrienoic acids by bovine coronary arteries
    Xiu Yu Yi
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, USA
    Am J Physiol Heart Circ Physiol 292:H2265-74. 2007
    ..Our results suggest a role of adrenic acid metabolites, specifically, DH-EETs as endothelium-derived hyperpolarizing factors in the coronary circulation...
  17. pmc 11,12,20-Trihydroxy-eicosa-8(Z)-enoic acid: a selective inhibitor of 11,12-EET-induced relaxations of bovine coronary and rat mesenteric arteries
    Ishfaq A Bukhari
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, Wisconsin 53225, USA
    Am J Physiol Heart Circ Physiol 302:H1574-83. 2012
    ..These results indicate that 11,12,20-THE8ZE is a selective antagonist of 11,12-EET relaxations and a useful pharmacological tool to elucidate the function of 11,12-EET in the cardiovascular system...
  18. pmc Soluble epoxide hydrolase contamination of specific catalase preparations inhibits epoxyeicosatrienoic acid vasodilation of rat renal arterioles
    Kathryn M Gauthier
    Department of Pharmacology and Cardiovascular Center, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA
    Am J Physiol Renal Physiol 301:F765-72. 2011
    ..However, some commercial catalase preparations are contaminated with sEH, and these contaminated preparations diminish the biological activity of H(2)O(2) and EETs...
  19. pmc 20-Iodo-14,15-epoxyeicosa-8(Z)-enoyl-3-azidophenylsulfonamide: photoaffinity labeling of a 14,15-epoxyeicosatrienoic acid receptor
    Yuenmu Chen
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, Wisconsin 53226, USA
    Biochemistry 50:3840-8. 2011
    ..This EET photoaffinity labeling method with a high signal-to-noise ratio may lead to new insights into the expression and regulation of the EET receptor...
  20. pmc Quantifying mitochondrial and plasma membrane potentials in intact pulmonary arterial endothelial cells based on extracellular disposition of rhodamine dyes
    Zhuohui Gan
    Department of Biomedical Engineering, Marquette University, Milwaukee, Wisconsin, USA
    Am J Physiol Lung Cell Mol Physiol 300:L762-72. 2011
    ....
  21. pmc Vascular pharmacology of epoxyeicosatrienoic acids
    Sandra L Pfister
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, Wisconsin, USA
    Adv Pharmacol 60:27-59. 2010
    ..Furthermore, the therapeutic potential of targeting the EET pathway in vascular disease will be highlighted...
  22. pmc 14,15-Dihydroxy-eicosa-5(Z)-enoic acid selectively inhibits 14,15-epoxyeicosatrienoic acid-induced relaxations in bovine coronary arteries
    Ishfaq A Bukhari
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Rd, Milwaukee, WI 53226, USA
    J Pharmacol Exp Ther 336:47-55. 2011
    ..The present study indicates that conversion of 14,15-EE5ZE to 14,15-DHE5ZE produces a 14,15-EET-selective antagonist that will be a useful pharmacological tool to identify EET receptor(s) and EET function in the cardiovascular system...
  23. pmc Inhibition of carcinoma cell motility by epoxyeicosatrienoic acid (EET) antagonists
    Kasem Nithipatikom
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, Wisconsin, USA
    Cancer Sci 101:2629-36. 2010
    ....
  24. pmc Epoxyeicosatrienoic acids and endothelium-dependent responses
    William B Campbell
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, USA
    Pflugers Arch 459:881-95. 2010
    ..Thus, the EETs are endothelium-derived hyperpolarizing factors mediating a portion of the relaxations to acetylcholine, bradykinin, shear stress, and cyclic stretch and regulate vascular tone in vitro and in vivo...
  25. pmc Lipid raft-redox signaling platforms in plasma membrane
    Fan Yi
    Department of Pharmacology and Toxicology, Medical College of Virginia Campus, Virginia Commonwealth University, Richmond, VA, USA
    Methods Mol Biol 580:93-107. 2009
    ..It is expected that information provided here will help readers to design necessary experiments in their studies on LR signaling platforms and redox regulation of cell function...
  26. pmc 20-125Iodo-14,15-epoxyeicosa-5(Z)-enoic acid: a high-affinity radioligand used to characterize the epoxyeicosatrienoic acid antagonist binding site
    Yuenmu Chen
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA
    J Pharmacol Exp Ther 331:1137-45. 2009
    ..The inhibition of binding by some cytochrome P450 inhibitors suggests an alternative mechanism of action for these drugs and could lead to new drug candidates that target the EET binding sites...
  27. pmc 14,15-Epoxyeicosa-5,8,11-trienoic acid (14,15-EET) surrogates containing epoxide bioisosteres: influence upon vascular relaxation and soluble epoxide hydrolase inhibition
    J R Falck
    Departments of Biochemistry and Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA
    J Med Chem 52:5069-75. 2009
    ..5 microM, IC(50) 16 nM). These data reveal differential structural parameters for the two pharmacophores that could assist the development of potent and specific in vivo drug candidates...
  28. pmc Evidence for role of epoxyeicosatrienoic acids in mediating ischemic preconditioning and postconditioning in dog
    Garrett J Gross
    Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, WI 53226, USA
    Am J Physiol Heart Circ Physiol 297:H47-52. 2009
    ..These results suggest a unique role for endogenous EETs in both IPC and POC...