Formation of Omega 3-Derived Electrophiles During Inflammation

Summary

Principal Investigator: FRANCISCO JOSE SCHOPFER
Abstract: DESCRIPTION (provided by applicant): Atherosclerosis accounts for three fourth of all deaths from cardiovascular disease and atherothrombotic diseases are projected to be the leading cause of death worldwide in 2020, representing a major health burden. Omega-3 (w-3) polyunsaturated fatty acids exert potent anti-inflammatory actions and beneficial cardiovascular effects. Although these properties are widely recognized, the underlying mechanisms remain largely unknown. An electrophilic mapping of lipids formed during macrophage activation showed that keto-derived w-3 fatty acids omega 3 derived-fatty acids were present at intracellular concentrations up to 200 nM. The formation of these species was dependent on COX-2, and administered aspirin increased the levels of these w-3 electrophilic fatty acid oxo-derivatives (EFOX). Recently, EFOX have been shown to be formed in human macrophages, neutrophils and animal tissues subjected to inflammation. Additionally, EFOX have been found to form glutathione and protein adducts and to modulate the inflammatory pathways through activation of Nrf2 pathway, inhibition of cytokine expression and reduction of inducible nitric oxide synthase levels. The hypothesis of this proposal is that EFOX are biologically relevant molecules with anti-atherogenic properties formed during inflammation and atherosclerosis that transduce the beneficial effects of w-3 fatty acids and aspirin through the electrophilic regulation of signaling pathways. To address this hypothesis we propose to chemically generate analytical tools (i.e. deuterated and labeled standards) to aid in the rigorous biological identification and quantitation of EFOX. These standards will be used for the analysis of biosynthetic pathways in macrophages and to assess the in vivo formation in a relevant murine model of atherosclerosis. The biochemical and signaling properties of these species and its targets will be analyzed using cellular models. Finally, the pharmacological effects of EFOX will be tested on a murine atherosclerosis model. At completion, we expect to have determined the different EFOX isomeric species and their in vivo relevance and formation. In addition, we expect to have gained a clear understanding of their electrophilic reactivities, modulation of phase 2 gene expression, in particular the Nrf2/KEAP1 pathway and their PPARg activation. It is expected that this proposal will lead to a better understanding of w-3 fatty acid effects and to improved pharmacological approaches to decrease atherosclerosis and its detrimental effects.
Funding Period: 2011-09-01 - 2016-07-31
more information: NIH RePORT

Top Publications

  1. pmc Novel keto-phospholipids are generated by monocytes and macrophages, detected in cystic fibrosis, and activate peroxisome proliferator-activated receptor-γ
    Victoria J Hammond
    School of Medicine, Cardiff University, Heath Park Campus, Cardiff CF14 4XN, United Kingdom
    J Biol Chem 287:41651-66. 2012
  2. pmc Olives and olive oil are sources of electrophilic fatty acid nitroalkenes
    Marco Fazzari
    Department of Pharmacology and Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, United States of America Fondazione Ri MED, Palermo, Italy
    PLoS ONE 9:e84884. 2014
  3. pmc Biomimetic nitration of conjugated linoleic acid: formation and characterization of naturally occurring conjugated nitrodienes
    Steven R Woodcock
    Department of Pharmacology and Chemical Biology, University of Pittsburgh Pittsburgh, Pennsylvania 15261, United States
    J Org Chem 79:25-33. 2014
  4. pmc Modulation of nitro-fatty acid signaling: prostaglandin reductase-1 is a nitroalkene reductase
    Dario A Vitturi
    Department of Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA
    J Biol Chem 288:25626-37. 2013
  5. pmc Inhibition of Mycobacterium tuberculosis PknG by non-catalytic rubredoxin domain specific modification: reaction of an electrophilic nitro-fatty acid with the Fe-S center
    Magdalena Gil
    Unidad de Bioquímica y Proteómica Analíticas, Institut Pasteur de Montevideo, Uruguay Unidad de Bioquímica y Proteómica Analíticas, Instituto de Investigaciones Biologicas Clemente Estable, Ministerio de Educación y Cultura, Uruguay
    Free Radic Biol Med 65:150-61. 2013
  6. pmc Synaptic Zn2+ inhibits neurotransmitter release by promoting endocannabinoid synthesis
    Tamara Perez-Rosello
    Department of Otolaryngology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261, USA
    J Neurosci 33:9259-72. 2013
  7. pmc Characterization and quantification of endogenous fatty acid nitroalkene metabolites in human urine
    Sonia R Salvatore
    Department of Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, PA 15261, USA
    J Lipid Res 54:1998-2009. 2013
  8. pmc Electrophilic nitro-fatty acids inhibit vascular inflammation by disrupting LPS-dependent TLR4 signalling in lipid rafts
    Luis Villacorta
    Cardiovascular Center, Department of Internal Medicine, University of Michigan Medical Center, North Campus Research Complex Building 26 Room 355S, 2800 Plymouth Road, Ann Arbor, MI, USA
    Cardiovasc Res 98:116-24. 2013
  9. pmc Conjugated linoleic acid is a preferential substrate for fatty acid nitration
    Gustavo Bonacci
    Department of Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA
    J Biol Chem 287:44071-82. 2012
  10. pmc Generation and dietary modulation of anti-inflammatory electrophilic omega-3 fatty acid derivatives
    Chiara Cipollina
    Fondazione Ri MED, Palermo, Italy Department of Pharmacology and Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, United States of America
    PLoS ONE 9:e94836. 2014

Research Grants

  1. PATHOPHYSIOLOGY OF THE ENDOTHELIUM
    Francis W Luscinskas; Fiscal Year: 2013
  2. Thrombus Formation and Antithrombotic Intervention
    John H Griffin; Fiscal Year: 2013
  3. Diverse Roles of Reactive Oxygen Species and Inflammation in Vascular Disease
    Kathy K Griendling; Fiscal Year: 2013
  4. Endothelial Cell Phenotypes in Health and Disease
    William C Aird; Fiscal Year: 2013
  5. Cardiac Fibrillation: Mechanisms and Therapy
    James N Weiss; Fiscal Year: 2013
  6. DEVELOPMENT AND CONTROL OF PULMONARY ALVEOLAR STABILITY
    Samuel Hawgood; Fiscal Year: 2013
  7. Sparstolonin B as an anti-atherogenic agent
    Daping Fan; Fiscal Year: 2013
  8. Ether Lipids, Elcosanoids, and Lung Cell Pathophysiology
    CHRISTINA CARROLL LESLIE; Fiscal Year: 2013
  9. Pathophysiology of Alveolar Epithelial Lung Injury
    Jacob I Sznajder; Fiscal Year: 2013
  10. HORMONAL REGULATION OF BLOOD PRESSURE
    Michal Laniado Schwartzman; Fiscal Year: 2013

Detail Information

Publications10

  1. pmc Novel keto-phospholipids are generated by monocytes and macrophages, detected in cystic fibrosis, and activate peroxisome proliferator-activated receptor-γ
    Victoria J Hammond
    School of Medicine, Cardiff University, Heath Park Campus, Cardiff CF14 4XN, United Kingdom
    J Biol Chem 287:41651-66. 2012
    ..The lipids are a new family of bioactive mediators from the 12/15-LOX pathway that may contribute to its known anti-inflammatory actions in vivo...
  2. pmc Olives and olive oil are sources of electrophilic fatty acid nitroalkenes
    Marco Fazzari
    Department of Pharmacology and Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, United States of America Fondazione Ri MED, Palermo, Italy
    PLoS ONE 9:e84884. 2014
    ..Since NO2-FA instigate adaptive anti-inflammatory gene expression and metabolic responses, these redox-derived metabolites may contribute to the cardiovascular benefits associated with the Mediterranean diet. ..
  3. pmc Biomimetic nitration of conjugated linoleic acid: formation and characterization of naturally occurring conjugated nitrodienes
    Steven R Woodcock
    Department of Pharmacology and Chemical Biology, University of Pittsburgh Pittsburgh, Pennsylvania 15261, United States
    J Org Chem 79:25-33. 2014
    ..This represents one of the few descriptions of naturally occurring conjugated nitro dienes (in particular, 1-nitro 1,3-diene), an unusual and highly reactive motif with few biological examples extant. ..
  4. pmc Modulation of nitro-fatty acid signaling: prostaglandin reductase-1 is a nitroalkene reductase
    Dario A Vitturi
    Department of Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA
    J Biol Chem 288:25626-37. 2013
    ..These results situate PtGR-1 as a critical modulator of both the steady state levels and signaling activities of fatty acid nitroalkenes in vivo. ..
  5. pmc Inhibition of Mycobacterium tuberculosis PknG by non-catalytic rubredoxin domain specific modification: reaction of an electrophilic nitro-fatty acid with the Fe-S center
    Magdalena Gil
    Unidad de Bioquímica y Proteómica Analíticas, Institut Pasteur de Montevideo, Uruguay Unidad de Bioquímica y Proteómica Analíticas, Instituto de Investigaciones Biologicas Clemente Estable, Ministerio de Educación y Cultura, Uruguay
    Free Radic Biol Med 65:150-61. 2013
    ..These results affirm the relevance of the Rbx domain as a target for PknG inhibition and support that electrophilic lipid reactions of Rbx-Cys may represent a new drug strategy for specific PknG inhibition. ..
  6. pmc Synaptic Zn2+ inhibits neurotransmitter release by promoting endocannabinoid synthesis
    Tamara Perez-Rosello
    Department of Otolaryngology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261, USA
    J Neurosci 33:9259-72. 2013
    ..We reveal a previously unknown action of synaptic Zn(2+): synaptic Zn(2+) inhibits glutamate release by promoting 2-AG synthesis...
  7. pmc Characterization and quantification of endogenous fatty acid nitroalkene metabolites in human urine
    Sonia R Salvatore
    Department of Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, PA 15261, USA
    J Lipid Res 54:1998-2009. 2013
    ..The measurement of this class of mediators can constitute a sensitive noninvasive index of metabolic and inflammatory status...
  8. pmc Electrophilic nitro-fatty acids inhibit vascular inflammation by disrupting LPS-dependent TLR4 signalling in lipid rafts
    Luis Villacorta
    Cardiovascular Center, Department of Internal Medicine, University of Michigan Medical Center, North Campus Research Complex Building 26 Room 355S, 2800 Plymouth Road, Ann Arbor, MI, USA
    Cardiovasc Res 98:116-24. 2013
    ....
  9. pmc Conjugated linoleic acid is a preferential substrate for fatty acid nitration
    Gustavo Bonacci
    Department of Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA
    J Biol Chem 287:44071-82. 2012
    ..These results affirm that metabolic and inflammatory reactions yield electrophilic products that can modulate adaptive cell signaling mechanisms...
  10. pmc Generation and dietary modulation of anti-inflammatory electrophilic omega-3 fatty acid derivatives
    Chiara Cipollina
    Fondazione Ri MED, Palermo, Italy Department of Pharmacology and Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, United States of America
    PLoS ONE 9:e94836. 2014
    ..Herein, the endogenous generation of ω-3 PUFAs electrophilic ketone derivatives and their hydroxy precursors was evaluated in human neutrophils. In addition, their dietary modulation was assessed through a randomized clinical trial...

Research Grants30

  1. PATHOPHYSIOLOGY OF THE ENDOTHELIUM
    Francis W Luscinskas; Fiscal Year: 2013
    ..g., heart attacks and strokes), as well as other organs and tissues of the body. These mechanistic insights may help identify novel therapeutic targets for the treatment of a broad spectrum of inflammatory diseases. ..
  2. Thrombus Formation and Antithrombotic Intervention
    John H Griffin; Fiscal Year: 2013
    ..New knowledge will contribute to improving prevention, diagnosis and treatment of relevant diseases related to thrombosis. ..
  3. Diverse Roles of Reactive Oxygen Species and Inflammation in Vascular Disease
    Kathy K Griendling; Fiscal Year: 2013
    ..Ultimately, this research may establish new unifying concepts linking conditions that alter vascular oxidant stress and inflammation to the molecular processes underlying vasculopathies. (End of Abstract) ..
  4. Endothelial Cell Phenotypes in Health and Disease
    William C Aird; Fiscal Year: 2013
    ..Core C ("Gene Targeting Core";William C. Aird, Core Leader) provides the necessary tools for targeting the Hprt locus and the loci of endogenous genes in ES cells and mice. ..
  5. Cardiac Fibrillation: Mechanisms and Therapy
    James N Weiss; Fiscal Year: 2013
    ..Together, these studies will provide critical groundwork necessary to develop and advance novel therapies for this major complication and cause of mortality from heart disease. ..
  6. DEVELOPMENT AND CONTROL OF PULMONARY ALVEOLAR STABILITY
    Samuel Hawgood; Fiscal Year: 2013
    ..abstract_text> ..
  7. Sparstolonin B as an anti-atherogenic agent
    Daping Fan; Fiscal Year: 2013
    ..The confirmation of the hypothesis will usher the development of SsnB as a new anti-atherogenic agent;it will also provide a pharmacological evidence for the causal role of TLR2 and TLR4 signaling in atherogenesis. ..
  8. Ether Lipids, Elcosanoids, and Lung Cell Pathophysiology
    CHRISTINA CARROLL LESLIE; Fiscal Year: 2013
    ..By using multidisciplinary approaches, we will determine the structural identity of lipid mediators, the molecular mechanisms involved in their production and how they function to regulate lung responses. ..
  9. Pathophysiology of Alveolar Epithelial Lung Injury
    Jacob I Sznajder; Fiscal Year: 2013
    ..The insights gained from the data generated from these studies will provide novel molecular targets for the development of new therapeutic strategies to treat patients with lung injury. ..
  10. HORMONAL REGULATION OF BLOOD PRESSURE
    Michal Laniado Schwartzman; Fiscal Year: 2013
    ..ular tone, in the pathophysiology of hypertension and cardiovascular disease. ..
  11. Inflammatory responses of vascular cells
    Paul L Fox; Fiscal Year: 2013
    ..abstract_text> ..
  12. Resolution Mechanisms in Acute Inflammation: Resolution Pharmacology
    CHARLES NICHOLAS SERHAN; Fiscal Year: 2013
    ..Selected synthetic SPM will be scaled-up for demonstration of their unique mode of action in vivo in a resolution pharmacology core using experimental disease models. Our broad goal is to bring forth new treatments in resolution. ..
  13. Structural bases of the functions of RNA-protein machines
    THOMAS ARTHUR STEITZ; Fiscal Year: 2013
    ..Also of interest will be the ways in which the structures and properties of RNA molecules can be utilized to carry out various biological functions often analogous to those performed by proteins. ..
  14. Redox-Derived Pulmonary Anti-Inflammatory Mediators
    BRUCE ALAN FREEMAN; Fiscal Year: 2013
    ....
  15. Neurohumoral control of veins in hypertension
    Gregory D Fink; Fiscal Year: 2013
    ..This project tests the idea that altered structure or function of veins also may cause hypertension, and that it may be possible to treat hypertension using drugs that affect veins. ..
  16. Improving Cardiac Function After Myocardial Infarction
    Steven R Houser; Fiscal Year: 2013
    ..A gene vector core will generate AAV6 vectors with novel therapeutics for testing in the pig Ml model. An administrative core will ensure data sharing and effective use of all resources. ..
  17. INTEGRATED MECHANISMS OF CARDIAC MALADAPTATION
    R John Solaro; Fiscal Year: 2013
    ..Studies proposed here offer the potential for novel diagnostic procedures early in the progression of the disorders, and targets for novel therapies. (End of Abstract) ..
  18. Role of Eosinophils in Airway Inflammation and Remodeling
    Nizar N Jarjour; Fiscal Year: 2013
    ..Given the prominence of eosinophilic inflammation in a significant proportion of severe asthma patients, these advances will have direct implications for the patients most affected by this very common illness. ..
  19. The Center for Native and Pacific Health Disparities Research
    MARJORIE K LEIMOMI MALA MAU; Fiscal Year: 2013
    ..5) To prepare and empower our diverse Native and Pacific People communities to take ownership of their own health and wellness. ..