Nomura DK


Full name : Nomura Daniel K

First name : Daniel K

Mail : Department of Chemical Physiology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037

Zip Code :

City :

Country : USA

Email :

Phone :

Fax :

Website :

Directory :

References (37)

Title : Enzyme promiscuity drives branched-chain fatty acid synthesis in adipose tissues - Wallace_2018_Nat.Chem.Biol_14_1021
Author(s) : Wallace M , Green CR , Roberts LS , Lee YM , McCarville JL , Sanchez-Gurmaches J , Meurs N , Gengatharan JM , Hover JD , Phillips SA , Ciaraldi TP , Guertin DA , Cabrales P , Ayres JS , Nomura DK , Loomba R , Metallo CM
Ref : Nat Chemical Biology , 14 :1021 , 2018
Abstract : Fatty acid synthase (FASN) predominantly generates straight-chain fatty acids using acetyl-CoA as the initiating substrate. However, monomethyl branched-chain fatty acids (mmBCFAs) are also present in mammals but are thought to be primarily diet derived. Here we demonstrate that mmBCFAs are de novo synthesized via mitochondrial BCAA catabolism, exported to the cytosol by adipose-specific expression of carnitine acetyltransferase (CrAT), and elongated by FASN. Brown fat exhibits the highest BCAA catabolic and mmBCFA synthesis fluxes, whereas these lipids are largely absent from liver and brain. mmBCFA synthesis is also sustained in the absence of microbiota. We identify hypoxia as a potent suppressor of BCAA catabolism that decreases mmBCFA synthesis in obese adipose tissue, such that mmBCFAs are significantly decreased in obese animals. These results identify adipose tissue mmBCFA synthesis as a novel link between BCAA metabolism and lipogenesis, highlighting roles for CrAT and FASN promiscuity influencing acyl-chain diversity in the lipidome.
ESTHER : Wallace_2018_Nat.Chem.Biol_14_1021
PubMedSearch : Wallace_2018_Nat.Chem.Biol_14_1021
PubMedID: 30327559
Gene_locus related to this paper: human-FASN

Title : Human Carboxylesterase 2 Reverses Obesity-Induced Diacylglycerol Accumulation and Glucose Intolerance - Ruby_2017_Cell.Rep_18_636
Author(s) : Ruby MA , Massart J , Hunerdosse DM , Schonke M , Correia JC , Louie SM , Ruas JL , Naslund E , Nomura DK , Zierath JR
Ref : Cell Rep , 18 :636 , 2017
Abstract : Serine hydrolases are a large family of multifunctional enzymes known to influence obesity. Here, we performed activity-based protein profiling to assess the functional level of serine hydrolases in liver biopsies from lean and obese humans in order to gain mechanistic insight into the pathophysiology of metabolic disease. We identified reduced hepatic activity of carboxylesterase 2 (CES2) and arylacetamide deacetylase (AADAC) in human obesity. In primary human hepatocytes, CES2 knockdown impaired glucose storage and lipid oxidation. In mice, obesity reduced CES2, whereas adenoviral delivery of human CES2 reversed hepatic steatosis, improved glucose tolerance, and decreased inflammation. Lipidomic analysis identified a network of CES2-regulated lipids altered in human and mouse obesity. CES2 possesses triglyceride and diacylglycerol lipase activities and displayed an inverse correlation with HOMA-IR and hepatic diacylglycerol concentrations in humans. Thus, decreased CES2 is a conserved feature of obesity and plays a causative role in the pathogenesis of obesity-related metabolic disturbances.
ESTHER : Ruby_2017_Cell.Rep_18_636
PubMedSearch : Ruby_2017_Cell.Rep_18_636
PubMedID: 28099843

Title : Lipases and their inhibitors in health and disease - Nomura_2016_Chem.Biol.Interact_259_211
Author(s) : Nomura DK , Casida JE
Ref : Chemico-Biological Interactions , 259 :211 , 2016
Abstract : Lipids play diverse and important biological roles including maintaining cellular integrity, storing fat for energy, acting as signaling molecules, and forming microdomains to support membrane protein signaling. Altering the levels of specific lipid species through activating or inactivating their biosynthetic or degradative pathways has been shown to provide either therapeutic benefit or cause disease. This review focuses on the functional, therapeutic, and (patho)physiological roles of lipases within the serine hydrolase superfamily and their inhibitors, with particular emphasis on the pharmacological tools, drugs, and environmental chemicals that inhibit these lipases.
ESTHER : Nomura_2016_Chem.Biol.Interact_259_211
PubMedSearch : Nomura_2016_Chem.Biol.Interact_259_211
PubMedID: 27067293

Title : Mapping proteome-wide interactions of reactive chemicals using chemoproteomic platforms - Counihan_2016_Curr.Opin.Chem.Biol_30_68
Author(s) : Counihan JL , Ford B , Nomura DK
Ref : Curr Opin Chemical Biology , 30 :68 , 2016
Abstract : A large number of pharmaceuticals, endogenous metabolites, and environmental chemicals act through covalent mechanisms with protein targets. Yet, their specific interactions with the proteome still remain poorly defined for most of these reactive chemicals. Deciphering direct protein targets of reactive small-molecules is critical in understanding their biological action, off-target effects, potential toxicological liabilities, and development of safer and more selective agents. Chemoproteomic technologies have arisen as a powerful strategy that enable the assessment of proteome-wide interactions of these irreversible agents directly in complex biological systems. We review here several chemoproteomic strategies that have facilitated our understanding of specific protein interactions of irreversibly-acting pharmaceuticals, endogenous metabolites, and environmental electrophiles to reveal novel pharmacological, biological, and toxicological mechanisms.
ESTHER : Counihan_2016_Curr.Opin.Chem.Biol_30_68
PubMedSearch : Counihan_2016_Curr.Opin.Chem.Biol_30_68
PubMedID: 26647369

Title : Mapping Proteome-Wide Targets of Environmental Chemicals Using Reactivity-Based Chemoproteomic Platforms - Medina-Cleghorn_2015_Chem.Biol_22_1394
Author(s) : Medina-Cleghorn D , Bateman LA , Ford B , Heslin A , Fisher KJ , Dalvie ED , Nomura DK
Ref : Chemical Biology , 22 :1394 , 2015
Abstract : We are exposed to a growing number of chemicals in our environment, most of which have not been characterized in terms of their toxicological potential or mechanisms. Here, we employ a chemoproteomic platform to map the cysteine reactivity of environmental chemicals using reactivity-based probes to mine for hyper-reactive hotspots across the proteome. We show that environmental contaminants such as monomethylarsonous acid and widely used pesticides such as chlorothalonil and chloropicrin possess common reactivity with a distinct set of proteins. Many of these proteins are involved in key metabolic processes, suggesting that these targets may be particularly sensitive to environmental electrophiles. We show that the widely used fungicide chlorothalonil specifically inhibits several metabolic enzymes involved in fatty acid metabolism and energetics, leading to dysregulated lipid metabolism in mice. Our results underscore the utility of using reactivity-based chemoproteomic platforms to uncover novel mechanistic insights into the toxicity of environmental chemicals.
ESTHER : Medina-Cleghorn_2015_Chem.Biol_22_1394
PubMedSearch : Medina-Cleghorn_2015_Chem.Biol_22_1394
PubMedID: 26496688

Title : Monoacylglycerol Lipase Regulates Fever Response - Sanchez-Alavez_2015_PLoS.One_10_e0134437
Author(s) : Sanchez-Alavez M , Nguyen W , Mori S , Moroncini G , Viader A , Nomura DK , Cravatt BF , Conti B
Ref : PLoS ONE , 10 :e0134437 , 2015
Abstract : Cyclooxygenase inhibitors such as ibuprofen have been used for decades to control fever through reducing the levels of the pyrogenic lipid transmitter prostaglandin E2 (PGE2). Historically, phospholipases have been considered to be the primary generator of the arachidonic acid (AA) precursor pool for generating PGE2 and other eicosanoids. However, recent studies have demonstrated that monoacyglycerol lipase (MAGL), through hydrolysis of the endocannabinoid 2-arachidonoylglycerol, provides a major source of AA for PGE2 synthesis in the mammalian brain under basal and neuroinflammatory states. We show here that either genetic or pharmacological ablation of MAGL leads to significantly reduced fever responses in both centrally or peripherally-administered lipopolysaccharide or interleukin-1beta-induced fever models in mice. We also show that a cannabinoid CB1 receptor antagonist does not attenuate these anti-pyrogenic effects of MAGL inhibitors. Thus, much like traditional nonsteroidal anti-inflammatory drugs, MAGL inhibitors can control fever, but appear to do so through restricted control over prostaglandin production in the nervous system.
ESTHER : Sanchez-Alavez_2015_PLoS.One_10_e0134437
PubMedSearch : Sanchez-Alavez_2015_PLoS.One_10_e0134437
PubMedID: 26287872

Title : Multidimensional profiling platforms reveal metabolic dysregulation caused by organophosphorus pesticides - Medina-Cleghorn_2014_ACS.Chem.Biol_9_423
Author(s) : Medina-Cleghorn D , Heslin A , Morris PJ , Mulvihill MM , Nomura DK
Ref : ACS Chemical Biology , 9 :423 , 2014
Abstract : We are environmentally exposed to countless synthetic chemicals on a daily basis, with an increasing number of these chemical exposures linked to adverse health effects. However, our understanding of the (patho)physiological effects of these chemicals remains poorly understood, due in part to a general lack of effort to systematically and comprehensively identify the direct interactions of environmental chemicals with biological macromolecules in mammalian systems in vivo. Here, we have used functional chemoproteomic and metabolomic platforms to broadly identify direct enzyme targets that are inhibited by widely used organophosphorus (OP) pesticides in vivo in mice and to determine metabolic alterations that are caused by these chemicals. We find that these pesticides directly inhibit over 20 serine hydrolases in vivo leading to widespread disruptions in lipid metabolism. Through identifying direct biological targets of OP pesticides, we show heretofore unrecognized modes of toxicity that may be associated with these agents and underscore the utility of using multidimensional profiling approaches to obtain a more complete understanding of toxicities associated with environmental chemicals.
ESTHER : Medina-Cleghorn_2014_ACS.Chem.Biol_9_423
PubMedSearch : Medina-Cleghorn_2014_ACS.Chem.Biol_9_423
PubMedID: 24205821

Title : Organophosphorus flame retardants inhibit specific liver carboxylesterases and cause serum hypertriglyceridemia - Morris_2014_ACS.Chem.Biol_9_1097
Author(s) : Morris PJ , Medina-Cleghorn D , Heslin A , King SM , Orr J , Mulvihill MM , Krauss RM , Nomura DK
Ref : ACS Chemical Biology , 9 :1097 , 2014
Abstract : Humans are prevalently exposed to organophosphorus flame retardants (OPFRs) contained in consumer products and electronics, though their toxicological effects and mechanisms remain poorly understood. We show here that OPFRs inhibit specific liver carboxylesterases (Ces) and cause altered hepatic lipid metabolism. Ablation of the OPFR target Ces1g has been previously linked to dyslipidemia in mice. Consistent with OPFR inhibition of Ces1g, we also observe OPFR-induced serum hypertriglyceridemia in mice. Our findings suggest novel toxicities that may arise from OPFR exposure and highlight the utility of chemoproteomic and metabolomic platforms in the toxicological characterization of environmental chemicals.
ESTHER : Morris_2014_ACS.Chem.Biol_9_1097
PubMedSearch : Morris_2014_ACS.Chem.Biol_9_1097
PubMedID: 24597639

Title : Chemical genetics screening reveals KIAA1363 as a cytokine-lowering target - Hunerdosse_2014_ACS.Chem.Biol_9_2905
Author(s) : Hunerdosse DM , Morris PJ , Miyamoto DK , Fisher KJ , Bateman LA , Ghazaleh JR , Zhong S , Nomura DK
Ref : ACS Chemical Biology , 9 :2905 , 2014
Abstract : Inflammation is a hallmark of many human diseases, including pain, arthritis, atherosclerosis, obesity and diabetes, cancer, and neurodegenerative diseases. Although there are several successfully marketed small molecules anti-inflammatory drugs such as cyclooxygenase inhibitors and glucocorticoids, many of these compounds are also associated with various adverse cardiovascular or immunosuppressive side effects. Thus, identifying novel anti-inflammatory small molecules and their targets is critical for developing safer and more effective next-generation treatment strategies for inflammatory diseases. Here, we have conducted a chemical genetics screen to identify small molecules that suppress the release of the inflammatory cytokine TNFalpha from stimulated macrophages. We have used an enzyme class-directed chemical library for our screening efforts to facilitate subsequent target identification using activity-based protein profiling (ABPP). Using this strategy, we have found that KIAA1363 is a novel target for lowering key pro-inflammatory cytokines through affecting key ether lipid metabolism pathways. Our study highlights the application of combining chemical genetics with chemoproteomic and metabolomic approaches toward identifying and characterizing anti-inflammatory smal molecules and their targets.
ESTHER : Hunerdosse_2014_ACS.Chem.Biol_9_2905
PubMedSearch : Hunerdosse_2014_ACS.Chem.Biol_9_2905
PubMedID: 25343321
Gene_locus related to this paper: human-NCEH1

Title : Integrated phenotypic and activity-based profiling links Ces3 to obesity and diabetes - Dominguez_2014_Nat.Chem.Biol_10_113
Author(s) : Dominguez E , Galmozzi A , Chang JW , Hsu KL , Pawlak J , Li W , Godio C , Thomas J , Partida D , Niessen S , O'Brien PE , Russell AP , Watt MJ , Nomura DK , Cravatt BF , Saez E
Ref : Nat Chemical Biology , 10 :113 , 2014
Abstract : Phenotypic screening is making a comeback in drug discovery as the maturation of chemical proteomics methods has facilitated target identification for bioactive small molecules. A limitation of these approaches is that time-consuming genetic methods or other means are often required to determine the biologically relevant target (or targets) from among multiple protein-compound interactions that are typically detected. Here, we have combined phenotypic screening of a directed small-molecule library with competitive activity-based protein profiling to map and functionally characterize the targets of screening hits. Using this approach, we identify carboxylesterase 3 (Ces3, also known as Ces1d) as a primary molecular target of bioactive compounds that promote lipid storage in adipocytes. We further show that Ces3 activity is markedly elevated during adipocyte differentiation. Treatment of two mouse models of obesity-diabetes with a Ces3 inhibitor ameliorates multiple features of metabolic syndrome, illustrating the power of the described strategy to accelerate the identification and pharmacologic validation of new therapeutic targets.
ESTHER : Dominguez_2014_Nat.Chem.Biol_10_113
PubMedSearch : Dominguez_2014_Nat.Chem.Biol_10_113
PubMedID: 24362705
Gene_locus related to this paper: human-CES3 , mouse-Ces1d

Title : Monoacylglycerol lipase inhibitor JZL184 improves behavior and neural properties in Ts65Dn mice, a model of down syndrome - Lysenko_2014_PLoS.One_9_e114521
Author(s) : Lysenko LV , Kim J , Henry C , Tyrtyshnaia A , Kohnz RA , Madamba F , Simon GM , Kleschevnikova NE , Nomura DK , Ezekowitz RA , Kleschevnikov AM
Ref : PLoS ONE , 9 :e114521 , 2014
Abstract : Genetic alterations or pharmacological treatments affecting endocannabinoid signaling have profound effects on synaptic and neuronal properties and, under certain conditions, may improve higher brain functions. Down syndrome (DS), a developmental disorder caused by triplication of chromosome 21, is characterized by deficient cognition and inevitable development of the Alzheimer disease (AD) type pathology during aging. Here we used JZL184, a selective inhibitor of monoacylglycerol lipase (MAGL), to examine the effects of chronic MAGL inhibition on the behavioral, biochemical, and synaptic properties of aged Ts65Dn mice, a genetic model of DS. In both Ts65Dn mice and their normosomic (2N) controls, JZL184-treatment increased brain levels of 2-arachidonoylglycerol (2-AG) and decreased levels of its metabolites such as arachidonic acid, prostaglandins PGD2, PGE2, PGFalpha, and PGJ2. Enhanced spontaneous locomotor activity of Ts65Dn mice was reduced by the JZL184-treatement to the levels observed in 2N animals. Deficient long-term memory was also improved, while short-term and working types of memory were unaffected. Furthermore, reduced hippocampal long-term potentiation (LTP) was increased in the JZL184-treated Ts65Dn mice to the levels observed in 2N mice. Interestingly, changes in synaptic plasticity and behavior were not observed in the JZL184-treated 2N mice suggesting that the treatment specifically attenuated the defects in the trisomic animals. The JZL184-treatment also reduced the levels of Abeta40 and Abeta42, but had no effect on the levels of full length APP and BACE1 in both Ts65Dn and 2N mice. These data show that chronic MAGL inhibition improves the behavior and brain functions in a DS model suggesting that pharmacological targeting of MAGL may be considered as a perspective new approach for improving cognition in DS.
ESTHER : Lysenko_2014_PLoS.One_9_e114521
PubMedSearch : Lysenko_2014_PLoS.One_9_e114521
PubMedID: 25474204

Title : Therapeutic potential of monoacylglycerol lipase inhibitors - Mulvihill_2013_Life.Sci_92_492
Author(s) : Mulvihill MM , Nomura DK
Ref : Life Sciences , 92 :492 , 2013
Abstract : Marijuana and aspirin have been used for millennia to treat a wide range of maladies including pain and inflammation. Both cannabinoids, like marijuana, that exert anti-inflammatory action through stimulating cannabinoid receptors, and cyclooxygenase (COX) inhibitors, like aspirin, that suppress pro-inflammatory eicosanoid production have shown beneficial outcomes in mouse models of neurodegenerative diseases and cancer. Both cannabinoids and COX inhibitors, however, have untoward effects that discourage their chronic usage, including cognitive deficits and gastrointestinal toxicity, respectively. Recent studies have uncovered that the serine hydrolase monoacylglycerol lipase (MAGL) links the endocannabinoid and eicosanoid systems together through hydrolysis of the endocannabinoid 2-arachidonoylglycerol (2-AG) to provide the major arachidonic acid (AA) precursor pools for pro-inflammatory eicosanoid synthesis in specific tissues. Studies in recent years have shown that MAGL inhibitors elicit anti-nociceptive, anxiolytic, and anti-emetic responses and attenuate precipitated withdrawal symptoms in addiction paradigms through enhancing endocannabinoid signaling. MAGL inhibitors have also been shown to exert anti-inflammatory action in the brain and protect against neurodegeneration through lowering eicosanoid production. In cancer, MAGL inhibitors have been shown to have anti-cancer properties not only through modulating the endocannabinoid-eicosanoid network, but also by controlling fatty acid release for the synthesis of protumorigenic signaling lipids. Thus, MAGL serves as a critical node in simultaneously coordinating multiple lipid signaling pathways in both physiological and disease contexts. This review will discuss the diverse (patho)physiological roles of MAGL and the therapeutic potential of MAGL inhibitors in treating a vast array of complex human diseases.
ESTHER : Mulvihill_2013_Life.Sci_92_492
PubMedSearch : Mulvihill_2013_Life.Sci_92_492
PubMedID: 23142242

Title : Monoacylglycerol lipase controls endocannabinoid and eicosanoid signaling and hepatic injury in mice - Cao_2013_Gastroenterology_144_808
Author(s) : Cao Z , Mulvihill MM , Mukhopadhyay P , Xu H , Erdelyi K , Hao E , Holovac E , Hasko G , Cravatt BF , Nomura DK , Pacher P
Ref : Gastroenterology , 144 :808 , 2013
Abstract : BACKGROUND & AIMS: The endocannabinoid and eicosanoid lipid signaling pathways have important roles in inflammatory syndromes. Monoacylglycerol lipase (MAGL) links these pathways, hydrolyzing the endocannabinoid 2-arachidonoylglycerol to generate the arachidonic acid precursor pool for prostaglandin production. We investigated whether blocking MAGL protects against inflammation and damage from hepatic ischemia/reperfusion (I/R) and other insults.
METHODS: We analyzed the effects of hepatic I/R in mice given the selective MAGL inhibitor JZL184, in Mgll(-/-) mice, fatty acid amide hydrolase(-/-) mice, and in cannabinoid receptor type 1(-/-) (CB1-/-) and cannabinoid receptor type 2(-/-) (CB2-/-). Liver tissues were collected and analyzed, along with cultured hepatocytes and Kupffer cells. We measured endocannabinoids, eicosanoids, and markers of inflammation, oxidative stress, and cell death using molecular biology, biochemistry, and mass spectrometry analyses.
RESULTS: Wild-type mice given JZL184 and Mgll(-/-) mice were protected from hepatic I/R injury by a mechanism that involved increased endocannabinoid signaling via CB2 and reduced production of eicosanoids in the liver. JZL184 suppressed the inflammation and oxidative stress that mediate hepatic I/R injury. Hepatocytes were the major source of hepatic MAGL activity and endocannabinoid and eicosanoid production. JZL184 also protected from induction of liver injury by D-(+)-galactosamine and lipopolysaccharides or CCl4.
CONCLUSIONS: MAGL modulates hepatic injury via endocannabinoid and eicosanoid signaling; blockade of this pathway protects mice from liver injury. MAGL inhibitors might be developed to treat conditions that expose the liver to oxidative stress and inflammatory damage.
ESTHER : Cao_2013_Gastroenterology_144_808
PubMedSearch : Cao_2013_Gastroenterology_144_808
PubMedID: 23295443
Gene_locus related to this paper: human-MGLL

Title : Activity-based protein profiling of organophosphorus and thiocarbamate pesticides reveals multiple serine hydrolase targets in mouse brain - Nomura_2011_J.Agric.Food.Chem_59_2808
Author(s) : Nomura DK , Casida JE
Ref : Journal of Agricultural and Food Chemistry , 59 :2808 , 2011
Abstract : Organophosphorus (OP) and thiocarbamate (TC) agrochemicals are used worldwide as insecticides, herbicides, and fungicides, but their safety assessment in terms of potential off-targets remains incomplete. In this study, a chemoproteomic platform, termed activity-based protein profiling, was used to broadly define serine hydrolase targets in mouse brain of a panel of 29 OP and TC pesticides. Among the secondary targets identified, enzymes involved in the degradation of endocannabinoid signaling lipids, monoacylglycerol lipase, and fatty acid amide hydrolase were inhibited by several OP and TC pesticides. Blockade of these two enzymes led to elevations in brain endocannabinoid levels and dysregulated brain arachidonate metabolism. Other secondary targets include enzymes thought to also play important roles in the nervous system and unannotated proteins. This study reveals a multitude of secondary targets for OP and TC pesticides and underscores the utility of chemoproteomic platforms in gaining insights into biochemical pathways that are perturbed by these toxicants.
ESTHER : Nomura_2011_J.Agric.Food.Chem_59_2808
PubMedSearch : Nomura_2011_J.Agric.Food.Chem_59_2808
PubMedID: 21341672

Title : Inhibition of monoacylglycerol lipase attenuates nonsteroidal anti-inflammatory drug-induced gastric hemorrhages in mice - Kinsey_2011_J.Pharmacol.Exp.Ther_338_795
Author(s) : Kinsey SG , Nomura DK , O'Neal ST , Long JZ , Mahadevan A , Cravatt BF , Grider JR , Lichtman AH
Ref : Journal of Pharmacology & Experimental Therapeutics , 338 :795 , 2011
Abstract : Nonsteroidal anti-inflammatory drugs (NSAIDs) are commonly used analgesics, but can cause gastric and esophageal hemorrhages, erosion, and ulceration. The endogenous cannabinoid (endocannabinoid; eCB) system possesses several potential targets to reduce gastric inflammatory states, including cannabinoid receptor type 1 (CB(1)), cannabinoid receptor type 2 (CB(2)), and enzymes that regulate the eCB ligands 2-arachidonoylglycerol (2-AG) and N-arachidonoyl ethanolamine (anandamide; AEA). In the presented study, we tested whether 4-nitrophenyl 4-(dibenzo[d][1,3]dioxol-5-yl(hydroxy)methyl)piperidine-1-carboxylate (JZL184), a selective inhibitor of the primary catabolic enzyme of 2-AG, monoacylglycerol lipase (MAGL), would protect against NSAID-induced gastric damage. Food-deprived mice administered the nonselective cyclooxygenase inhibitor diclofenac sodium displayed gastric hemorrhages and increases in proinflammatory cytokines. JZL184, the proton pump inhibitor omeprazole (positive control), or the primary constituent of marijuana, delta(9)-tetrahydrocannabinol (THC), significantly prevented diclofenac-induced gastric hemorrhages. JZL184 also increased stomach levels of 2-AG, but had no effect on AEA, arachidonic acid, or the prostaglandins E(2) and D(2). MAGL inhibition fully blocked diclofenac-induced increases in gastric levels of proinflammatory cytokines interleukin (IL)-1beta, IL-6, tumor necrosis factor alpha, and granulocyte colony-stimulating factor, as well as IL-10. Pharmacological inhibition or genetic deletion of CB(1) or CB(2) revealed that the gastroprotective effects of JZL184 and THC were mediated via CB(1). The antihemorrhagic effects of JZL184 persisted with repeated administration, indicating a lack of tolerance. These data indicate that increasing 2-AG protects against gastric damage induced by NSAIDs, and its primary catabolic enzyme MAGL offers a promising target for the development of analgesic therapeutics possessing gastroprotective properties.
ESTHER : Kinsey_2011_J.Pharmacol.Exp.Ther_338_795
PubMedSearch : Kinsey_2011_J.Pharmacol.Exp.Ther_338_795
PubMedID: 21659471

Title : Endocannabinoid hydrolysis generates brain prostaglandins that promote neuroinflammation - Nomura_2011_Science_334_809
Author(s) : Nomura DK , Morrison BE , Blankman JL , Long JZ , Kinsey SG , Marcondes MC , Ward AM , Hahn YK , Lichtman AH , Conti B , Cravatt BF
Ref : Science , 334 :809 , 2011
Abstract : Phospholipase A(2)(PLA(2)) enzymes are considered the primary source of arachidonic acid for cyclooxygenase (COX)-mediated biosynthesis of prostaglandins. Here, we show that a distinct pathway exists in brain, where monoacylglycerol lipase (MAGL) hydrolyzes the endocannabinoid 2-arachidonoylglycerol to generate a major arachidonate precursor pool for neuroinflammatory prostaglandins. MAGL-disrupted animals show neuroprotection in a parkinsonian mouse model. These animals are spared the hemorrhaging caused by COX inhibitors in the gut, where prostaglandins are instead regulated by cytosolic PLA(2). These findings identify MAGL as a distinct metabolic node that couples endocannabinoid to prostaglandin signaling networks in the nervous system and suggest that inhibition of this enzyme may be a new and potentially safer way to suppress the proinflammatory cascades that underlie neurodegenerative disorders.
ESTHER : Nomura_2011_Science_334_809
PubMedSearch : Nomura_2011_Science_334_809
PubMedID: 22021672

Title : Blockade of endocannabinoid hydrolytic enzymes attenuates precipitated opioid withdrawal symptoms in mice - Ramesh_2011_J.Pharmacol.Exp.Ther_339_173
Author(s) : Ramesh D , Ross GR , Schlosburg JE , Owens RA , Abdullah RA , Kinsey SG , Long JZ , Nomura DK , Sim-Selley LJ , Cravatt BF , Akbarali HI , Lichtman AH
Ref : Journal of Pharmacology & Experimental Therapeutics , 339 :173 , 2011
Abstract : Delta(9)-Tetrahydrocannbinol (THC), the primary active constituent of Cannabis sativa, has long been known to reduce opioid withdrawal symptoms. Although THC produces most of its pharmacological actions through the activation of CB(1) and CB(2) cannabinoid receptors, the role these receptors play in reducing the variety of opioid withdrawal symptoms remains unknown. The endogenous cannabinoids, N-arachidonoylethanolamine (anandamide; AEA) and 2-arachidonylglycerol (2-AG), activate both cannabinoid receptors but are rapidly metabolized by fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL), respectively. The objective of this study was to test whether increasing AEA or 2-AG, via inhibition of their respective hydrolytic enzymes, reduces naloxone-precipitated morphine withdrawal symptoms in in vivo and in vitro models of opioid dependence. Morphine-dependent mice challenged with naloxone reliably displayed a profound withdrawal syndrome, consisting of jumping, paw tremors, diarrhea, and weight loss. THC and the MAGL inhibitor 4-nitrophenyl 4-(dibenzo[d][1,3]dioxol-5-yl(hydroxy)methyl)piperidine-1-carboxylate (JZL184) dose dependently reduced the intensity of most measures through the activation of CB(1) receptors. JZL184 also attenuated spontaneous withdrawal signs in morphine-dependent mice. The FAAH inhibitor N-(pyridin-3-yl)-4-(3-(5-(trifluoromethyl)pyridin-2-yloxy)benzyl)-piperdine-1-car boxamide (PF-3845) reduced the intensity of naloxone-precipitated jumps and paw flutters through the activation of CB(1) receptors but did not ameliorate incidence of diarrhea or weight loss. In the final series of experiments, we investigated whether JZL184 or PF-3845 would attenuate naloxone-precipitated contractions in morphine-dependent ilea. Both enzyme inhibitors attenuated the intensity of naloxone-induced contractions, although this model does not account mechanistically for the autonomic withdrawal responses (i.e., diarrhea) observed in vivo. These results indicate that endocannabinoid catabolic enzymes are promising targets to treat opioid dependence.
ESTHER : Ramesh_2011_J.Pharmacol.Exp.Ther_339_173
PubMedSearch : Ramesh_2011_J.Pharmacol.Exp.Ther_339_173
PubMedID: 21719468

Title : A potent and selective inhibitor of KIAA1363\/AADACL1 that impairs prostate cancer pathogenesis - Chang_2011_Chem.Biol_18_476
Author(s) : Chang JW , Nomura DK , Cravatt BF
Ref : Chemical Biology , 18 :476 , 2011
Abstract : Cancer cells show alterations in metabolism that support malignancy and disease progression. Prominent among these metabolic changes is elevations in neutral ether lipids (NELs). We have previously shown that the hydrolytic enzyme KIAA1363 (or AADACL1) is highly elevated in aggressive cancer cells, where it plays a key role in generating the monoalkylglycerol ether (MAGE) class of NELs. Here, we use activity-based protein profiling-guided medicinal chemistry to discover a highly potent and selective inhibitor of KIAA1363, the carbamate JW480. We show that JW480, and an shRNA probe that targets KIAA1363, reduce MAGEs and impair the migration, invasion, survival, and in vivo tumor growth of human prostate cancer cell lines. These findings indicate that the KIAA1363-MAGE pathway is important for prostate cancer pathogenesis and designate JW480 as a versatile pharmacological probe for disrupting this pro-tumorigenic metabolic pathway.
ESTHER : Chang_2011_Chem.Biol_18_476
PubMedSearch : Chang_2011_Chem.Biol_18_476
PubMedID: 21513884
Gene_locus related to this paper: human-NCEH1

Title : Acute overactive endocannabinoid signaling induces glucose intolerance, hepatic steatosis, and novel cannabinoid receptor 1 responsive genes - Ruby_2011_PLoS.One_6_e26415
Author(s) : Ruby MA , Nomura DK , Hudak CS , Barber A , Casida JE , Krauss RM
Ref : PLoS ONE , 6 :e26415 , 2011
Abstract : Endocannabinoids regulate energy balance and lipid metabolism by stimulating the cannabinoid receptor type 1 (CB1). Genetic deletion and pharmacological antagonism have shown that CB1 signaling is necessary for the development of obesity and related metabolic disturbances. However, the sufficiency of endogenously produced endocannabinoids to cause hepatic lipid accumulation and insulin resistance, independent of food intake, has not been demonstrated. Here, we show that a single administration of isopropyl dodecylfluorophosphonate (IDFP), perhaps the most potent pharmacological inhibitor of endocannabinoid degradation, increases hepatic triglycerides (TG) and induces insulin resistance in mice. These effects involve increased CB1 signaling, as they are mitigated by pre-administration of a CB1 antagonist (AM251) and in CB1 knockout mice. Despite the strong physiological effects of CB1 on hepatic lipid and glucose metabolism, little is known about the downstream targets responsible for these effects. To elucidate transcriptional targets of CB1 signaling, we performed microarrays on hepatic RNA isolated from DMSO (control), IDFP and AM251/IDFP-treated mice. The gene for the secreted glycoprotein lipocalin 2 (lcn2), which has been implicated in obesity and insulin resistance, was among those most responsive to alterations in CB1 signaling. The expression pattern of IDFP mice segregated from DMSO mice in hierarchal cluster analysis and AM251 pre-administration reduced (>50%) the majority (303 of 533) of the IDFP induced alterations. Pathway analysis revealed that IDFP altered expression of genes involved in lipid, fatty acid and steroid metabolism, the acute phase response, and amino acid metabolism in a CB1-dependent manner. PCR confirmed array results of key target genes in multiple independent experiments. Overall, we show that acute IDFP treatment induces hepatic TG accumulation and insulin resistance, at least in part through the CB1 receptor, and identify novel cannabinoid responsive genes.
ESTHER : Ruby_2011_PLoS.One_6_e26415
PubMedSearch : Ruby_2011_PLoS.One_6_e26415
PubMedID: 22073164

Title : Academic cross-fertilization by public screening yields a remarkable class of protein phosphatase methylesterase-1 inhibitors - Bachovchin_2011_Proc.Natl.Acad.Sci.U.S.A_108_6811
Author(s) : Bachovchin DA , Mohr JT , Speers AE , Wang C , Berlin JM , Spicer TP , Fernandez-Vega V , Chase P , Hodder PS , Schurer SC , Nomura DK , Rosen H , Fu GC , Cravatt BF
Ref : Proc Natl Acad Sci U S A , 108 :6811 , 2011
Abstract : National Institutes of Health (NIH)-sponsored screening centers provide academic researchers with a special opportunity to pursue small-molecule probes for protein targets that are outside the current interest of, or beyond the standard technologies employed by, the pharmaceutical industry. Here, we describe the outcome of an inhibitor screen for one such target, the enzyme protein phosphatase methylesterase-1 (PME-1), which regulates the methylesterification state of protein phosphatase 2A (PP2A) and is implicated in cancer and neurodegeneration. Inhibitors of PME-1 have not yet been described, which we attribute, at least in part, to a dearth of substrate assays compatible with high-throughput screening. We show that PME-1 is assayable by fluorescence polarization-activity-based protein profiling (fluopol-ABPP) and use this platform to screen the 300,000+ member NIH small-molecule library. This screen identified an unusual class of compounds, the aza-beta-lactams (ABLs), as potent (IC(50) values of approximately 10 nM), covalent PME-1 inhibitors. Interestingly, ABLs did not derive from a commercial vendor but rather an academic contribution to the public library. We show using competitive-ABPP that ABLs are exquisitely selective for PME-1 in living cells and mice, where enzyme inactivation leads to substantial reductions in demethylated PP2A. In summary, we have combined advanced synthetic and chemoproteomic methods to discover a class of ABL inhibitors that can be used to selectively perturb PME-1 activity in diverse biological systems. More generally, these results illustrate how public screening centers can serve as hubs to create spontaneous collaborative opportunities between synthetic chemistry and chemical biology labs interested in creating first-in-class pharmacological probes for challenging protein targets.
ESTHER : Bachovchin_2011_Proc.Natl.Acad.Sci.U.S.A_108_6811
PubMedSearch : Bachovchin_2011_Proc.Natl.Acad.Sci.U.S.A_108_6811
PubMedID: 21398589
Gene_locus related to this paper: human-PPME1

Title : Monoacylglycerol lipase regulates a fatty acid network that promotes cancer pathogenesis - Nomura_2010_Cell_140_49
Author(s) : Nomura DK , Long JZ , Niessen S , Hoover HS , Ng SW , Cravatt BF
Ref : Cell , 140 :49 , 2010
Abstract : Tumor cells display progressive changes in metabolism that correlate with malignancy, including development of a lipogenic phenotype. How stored fats are liberated and remodeled to support cancer pathogenesis, however, remains unknown. Here, we show that the enzyme monoacylglycerol lipase (MAGL) is highly expressed in aggressive human cancer cells and primary tumors, where it regulates a fatty acid network enriched in oncogenic signaling lipids that promotes migration, invasion, survival, and in vivo tumor growth. Overexpression of MAGL in nonaggressive cancer cells recapitulates this fatty acid network and increases their pathogenicity-phenotypes that are reversed by an MAGL inhibitor. Impairments in MAGL-dependent tumor growth are rescued by a high-fat diet, indicating that exogenous sources of fatty acids can contribute to malignancy in cancers lacking MAGL activity. Together, these findings reveal how cancer cells can co-opt a lipolytic enzyme to translate their lipogenic state into an array of protumorigenic signals. PAPERFLICK:
ESTHER : Nomura_2010_Cell_140_49
PubMedSearch : Nomura_2010_Cell_140_49
PubMedID: 20079333
Gene_locus related to this paper: human-MGLL , mouse-MGLL

Title : Chronic monoacylglycerol lipase blockade causes functional antagonism of the endocannabinoid system - Schlosburg_2010_Nat.Neurosci_13_1113
Author(s) : Schlosburg JE , Blankman JL , Long JZ , Nomura DK , Pan B , Kinsey SG , Nguyen PT , Ramesh D , Booker L , Burston JJ , Thomas EA , Selley DE , Sim-Selley LJ , Liu QS , Lichtman AH , Cravatt BF
Ref : Nat Neurosci , 13 :1113 , 2010
Abstract : Prolonged exposure to drugs of abuse, such as cannabinoids and opioids, leads to pharmacological tolerance and receptor desensitization in the nervous system. We found that a similar form of functional antagonism was produced by sustained inactivation of monoacylglycerol lipase (MAGL), the principal degradative enzyme for the endocannabinoid 2-arachidonoylglycerol. After repeated administration, the MAGL inhibitor JZL184 lost its analgesic activity and produced cross-tolerance to cannabinoid receptor (CB1) agonists in mice, effects that were phenocopied by genetic disruption of Mgll (encoding MAGL). Chronic MAGL blockade also caused physical dependence, impaired endocannabinoid-dependent synaptic plasticity and desensitized brain CB1 receptors. These data contrast with blockade of fatty acid amide hydrolase, an enzyme that degrades the other major endocannabinoid anandamide, which produced sustained analgesia without impairing CB1 receptors. Thus, individual endocannabinoids generate distinct analgesic profiles that are either sustained or transitory and associated with agonism and functional antagonism of the brain cannabinoid system, respectively.
ESTHER : Schlosburg_2010_Nat.Neurosci_13_1113
PubMedSearch : Schlosburg_2010_Nat.Neurosci_13_1113
PubMedID: 20729846

Title : Activity-based protein profiling for biochemical pathway discovery in cancer - Nomura_2010_Nat.Rev.Cancer_10_630
Author(s) : Nomura DK , Dix MM , Cravatt BF
Ref : Nat Rev Cancer , 10 :630 , 2010
Abstract : Large-scale profiling methods have uncovered numerous gene and protein expression changes that correlate with tumorigenesis. However, determining the relevance of these expression changes and which biochemical pathways they affect has been hindered by our incomplete understanding of the proteome and its myriad functions and modes of regulation. Activity-based profiling platforms enable both the discovery of cancer-relevant enzymes and selective pharmacological probes to perturb and characterize these proteins in tumour cells. When integrated with other large-scale profiling methods, activity-based proteomics can provide insight into the metabolic and signalling pathways that support cancer pathogenesis and illuminate new strategies for disease diagnosis and treatment.
ESTHER : Nomura_2010_Nat.Rev.Cancer_10_630
PubMedSearch : Nomura_2010_Nat.Rev.Cancer_10_630
PubMedID: 20703252

Title : Dual blockade of FAAH and MAGL identifies behavioral processes regulated by endocannabinoid crosstalk in vivo - Long_2009_Proc.Natl.Acad.Sci.U.S.A_106_20270
Author(s) : Long JZ , Nomura DK , Vann RE , Walentiny DM , Booker L , Jin X , Burston JJ , Sim-Selley LJ , Lichtman AH , Wiley JL , Cravatt BF
Ref : Proc Natl Acad Sci U S A , 106 :20270 , 2009
Abstract : Delta(9)-tetrahydrocannabinol (THC), the psychoactive component of marijuana, and other direct cannabinoid receptor (CB1) agonists produce a number of neurobehavioral effects in mammals that range from the beneficial (analgesia) to the untoward (abuse potential). Why, however, this full spectrum of activities is not observed upon pharmacological inhibition or genetic deletion of either fatty acid amide hydrolase (FAAH) or monoacylglycerol lipase (MAGL), enzymes that regulate the two major endocannabinoids anandamide (AEA) and 2-arachidonoylglycerol (2-AG), respectively, has remained unclear. Here, we describe a selective and efficacious dual FAAH/MAGL inhibitor, JZL195, and show that this agent exhibits broad activity in the tetrad test for CB1 agonism, causing analgesia, hypomotilty, and catalepsy. Comparison of JZL195 to specific FAAH and MAGL inhibitors identified behavioral processes that were regulated by a single endocannabinoid pathway (e.g., hypomotility by the 2-AG/MAGL pathway) and, interestingly, those where disruption of both FAAH and MAGL produced additive effects that were reversed by a CB1 antagonist. Falling into this latter category was drug discrimination behavior, where dual FAAH/MAGL blockade, but not disruption of either FAAH or MAGL alone, produced THC-like responses that were reversed by a CB1 antagonist. These data indicate that AEA and 2-AG signaling pathways interact to regulate specific behavioral processes in vivo, including those relevant to drug abuse, thus providing a potential mechanistic basis for the distinct pharmacological profiles of direct CB1 agonists and inhibitors of individual endocannabinoid degradative enzymes.
ESTHER : Long_2009_Proc.Natl.Acad.Sci.U.S.A_106_20270
PubMedSearch : Long_2009_Proc.Natl.Acad.Sci.U.S.A_106_20270
PubMedID: 19918051

Title : Characterization of monoacylglycerol lipase inhibition reveals differences in central and peripheral endocannabinoid metabolism - Long_2009_Chem.Biol_16_744
Author(s) : Long JZ , Nomura DK , Cravatt BF
Ref : Chemical Biology , 16 :744 , 2009
Abstract : Monoacylglycerol lipase (MAGL) is a principal degradative enzyme for the endocannabinoid 2-arachidonoylglycerol (2-AG). We recently reported a piperidine carbamate, JZL184, that inhibits MAGL with high potency and selectivity. Here, we describe a comprehensive mechanistic characterization of JZL184. We provide evidence that JZL184 irreversibly inhibits MAGL via carbamoylation of the enzyme's serine nucleophile. Functional proteomic analysis of mice treated with JZL184 revealed that this inhibitor maintains good selectivity for MAGL across a wide range of central and peripheral tissues. Interestingly, MAGL blockade produced marked, tissue-specific differences in monoglyceride metabolism, with brain showing the most dramatic elevations in 2-AG and peripheral tissues often showing greater changes in other monoglycerides. Collectively, these studies indicate that MAGL exerts tissue-dependent control over endocannabinoid and monoglyceride metabolism and designate JZL184 as a selective tool to characterize the functions of MAGL in vivo.
ESTHER : Long_2009_Chem.Biol_16_744
PubMedSearch : Long_2009_Chem.Biol_16_744
PubMedID: 19635411

Title : Monoacylglycerol lipase regulates 2-arachidonoylglycerol action and arachidonic acid levels - Nomura_2008_Bioorg.Med.Chem.Lett_18_5875
Author(s) : Nomura DK , Hudak CS , Ward AM , Burston JJ , Issa RS , Fisher KJ , Abood ME , Wiley JL , Lichtman AH , Casida JE
Ref : Bioorganic & Medicinal Chemistry Lett , 18 :5875 , 2008
Abstract : The structure-activity relationships of organophosphorus (OP) and organosulfur compounds were examined in vitro and in vivo as inhibitors of mouse brain monoacylglycerol lipase (MAGL) hydrolysis of 2-arachidonoylglycerol (2-AG) and agonist binding at the CB1 receptor. Several compounds showed exceptional potency toward MAGL activity with IC(50) values of 0.1-10 nM in vitro and high inhibition at 10mg/kg intraperitoneally in mice. We find for the first time that MAGL activity is a major in vivo determinant of 2-AG and arachidonic acid levels not only in brain but also in spleen, lung, and liver. Apparent direct OP inhibition of CB1 agonist binding may be due instead to metabolic stabilization of 2-AG in brain membranes as the actual inhibitor.
ESTHER : Nomura_2008_Bioorg.Med.Chem.Lett_18_5875
PubMedSearch : Nomura_2008_Bioorg.Med.Chem.Lett_18_5875
PubMedID: 18752948

Title : Organophosphate-sensitive lipases modulate brain lysophospholipids, ether lipids and endocannabinoids - Casida_2008_Chem.Biol.Interact_175_355
Author(s) : Casida JE , Nomura DK , Vose SC , Fujioka K
Ref : Chemico-Biological Interactions , 175 :355 , 2008
Abstract : Lipases play key roles in nearly all cells and organisms. Potent and selective inhibitors help to elucidate their physiological functions and associated metabolic pathways. Organophosphorus (OP) compounds are best known for their anticholinesterase properties but selectivity for lipases and other targets can also be achieved through structural optimization. This review considers several lipid systems in brain modulated by highly OP-sensitive lipases. Neuropathy target esterase (NTE) hydrolyzes lysophosphatidylcholine (lysoPC) as a preferred substrate. Gene deletion of NTE in mice is embryo lethal and the heterozygotes are hyperactive. NTE is very sensitive in vitro and in vivo to direct-acting OP delayed neurotoxicants and the related NTE-related esterase (NTE-R) is also inhibited in vivo. KIAA1363 hydrolyzes acetyl monoalkylglycerol ether (AcMAGE) of the platelet-activating factor (PAF) de novo biosynthetic pathway and is a marker of cancer cell invasiveness. It is also a detoxifying enzyme that hydrolyzes chlorpyrifos oxon (CPO) and some other potent insecticide metabolites. Monoacylglycerol lipase and fatty acid amide hydrolase regulate endocannabinoid levels with roles in motility, pain and memory. Inhibition of these enzymes in mice by OPs, such as isopropyl dodecylfluorophosphonate (IDFP), leads to dramatic elevation of brain endocannabinoids and distinct cannabinoid-dependent behavior. Hormone-sensitive lipase that hydrolyzes cholesteryl esters and diacylglycerols is a newly recognized in vivo CPO- and IDFP-target in brain. The OP chemotype can therefore be used in proteomic and metabolomic studies to further elucidate the biological function and toxicological significance of lipases in lipid metabolism. Only the first steps have been taken to achieve appropriate selective action for OP therapeutic agents.
ESTHER : Casida_2008_Chem.Biol.Interact_175_355
PubMedSearch : Casida_2008_Chem.Biol.Interact_175_355
PubMedID: 18495101
Gene_locus related to this paper: human-NCEH1 , mouse-Q8BLF1

Title : Dual roles of brain serine hydrolase KIAA1363 in ether lipid metabolism and organophosphate detoxification - Nomura_2008_Toxicol.Appl.Pharmacol_228_42
Author(s) : Nomura DK , Fujioka K , Issa RS , Ward AM , Cravatt BF , Casida JE
Ref : Toxicol Appl Pharmacol , 228 :42 , 2008
Abstract : Serine hydrolase KIAA1363 is an acetyl monoalkylglycerol ether (AcMAGE) hydrolase involved in tumor cell invasiveness. It is also an organophosphate (OP) insecticide-detoxifying enzyme. The key to understanding these dual properties was the use of KIAA1363 +/+ (wildtype) and -/- (gene deficient) mice to define the role of this enzyme in brain and other tissues and its effectiveness in vivo in reducing OP toxicity. KIAA1363 was the primary AcMAGE hydrolase in brain, lung, heart and kidney and was highly sensitive to inactivation by chlorpyrifos oxon (CPO) (IC50 2 nM) [the bioactivated metabolite of the major insecticide chlorpyrifos (CPF)]. Although there was no difference in hydrolysis product monoalkylglycerol ether (MAGE) levels in +/+ and -/- mouse brains in vivo, isopropyl dodecylfluorophosphonate (30 mg/kg) and CPF (100 mg/kg) resulted in 23-51% decrease in brain MAGE levels consistent with inhibition of AcMAGE hydrolase activity. On incubating +/+ and -/- brain membranes with AcMAGE and cytidine-5'-diphosphocholine, the absence of KIAA1363 activity dramatically increased de novo formation of platelet-activating factor (PAF) and lyso-PAF, signifying that metabolically-stabilized AcMAGE can be converted to this bioactive lipid in brain. On considering detoxification, KIAA1363 -/- mice were significantly more sensitive than +/+ mice to ip-administered CPF (100 mg/kg) and parathion (10 mg/kg) with increased tremoring and mortality that correlated for CPF with greater brain acetylcholinesterase inhibition. Docking AcMAGE and CPO in a KIAA1363 active site model showed similar positioning of their acetyl and trichloropyridinyl moieties, respectively. This study establishes the relevance of KIAA1363 in ether lipid metabolism and OP detoxification.
ESTHER : Nomura_2008_Toxicol.Appl.Pharmacol_228_42
PubMedSearch : Nomura_2008_Toxicol.Appl.Pharmacol_228_42
PubMedID: 18164358
Gene_locus related to this paper: human-NCEH1

Title : Overactive endocannabinoid signaling impairs apolipoprotein E-mediated clearance of triglyceride-rich lipoproteins - Ruby_2008_Proc.Natl.Acad.Sci.U.S.A_105_14561
Author(s) : Ruby MA , Nomura DK , Hudak CS , Mangravite LM , Chiu S , Casida JE , Krauss RM
Ref : Proc Natl Acad Sci U S A , 105 :14561 , 2008
Abstract : The endocannabinoid (EC) system regulates food intake and energy metabolism. Cannabinoid receptor type 1 (CB1) antagonists show promise in the treatment of obesity and its metabolic consequences. Although the reduction in adiposity resulting from therapy with CB1 antagonists may not account fully for the concomitant improvements in dyslipidemia, direct effects of overactive EC signaling on plasma lipoprotein metabolism have not been documented. The present study used a chemical approach to evaluate the direct effects of increased EC signaling in mice by inducing acute elevations of endogenously produced cannabinoids through pharmacological inhibition of their enzymatic hydrolysis by isopropyl dodecylfluorophosphonate (IDFP). Acute IDFP treatment increased plasma levels of triglyceride (TG) (2.0- to 3.1-fold) and cholesterol (1.3- to 1.4-fold) in conjunction with an accumulation in plasma of apolipoprotein (apo)E-depleted TG-rich lipoproteins. These changes did not occur in either CB1-null or apoE-null mice, were prevented by pretreatment with CB1 antagonists, and were not associated with reduced hepatic apoE gene expression. Although IDFP treatment increased hepatic mRNA levels of lipogenic genes (Srebp1 and Fas), there was no effect on TG secretion into plasma. Instead, IDFP treatment impaired clearance of an intravenously administered TG emulsion, despite increased postheparin lipoprotein lipase activity. Therefore, overactive EC signaling elicits an increase in plasma triglyceride levels associated with reduced plasma TG clearance and an accumulation in plasma of apoE-depleted TG-rich lipoproteins. These findings suggest a role of CB1 activation in the pathogenesis of obesity-related hypertriglyceridemia and underscore the potential efficacy of CB1 antagonists in treating metabolic disease.
ESTHER : Ruby_2008_Proc.Natl.Acad.Sci.U.S.A_105_14561
PubMedSearch : Ruby_2008_Proc.Natl.Acad.Sci.U.S.A_105_14561
PubMedID: 18794527

Title : Activation of the endocannabinoid system by organophosphorus nerve agents - Nomura_2008_Nat.Chem.Biol_4_373
Author(s) : Nomura DK , Blankman JL , Simon GM , Fujioka K , Issa RS , Ward AM , Cravatt BF , Casida JE
Ref : Nat Chemical Biology , 4 :373 , 2008
Abstract : Delta(9)-tetrahydrocannabinol (THC), the psychoactive ingredient of marijuana, has useful medicinal properties but also undesirable side effects. The brain receptor for THC, CB(1), is also activated by the endogenous cannabinoids anandamide and 2-arachidonylglycerol (2-AG). Augmentation of endocannabinoid signaling by blockade of their metabolism may offer a more selective pharmacological approach compared with CB(1) agonists. Consistent with this premise, inhibitors of the anandamide-degrading enzyme fatty acid amide hydrolase (FAAH) produce analgesic and anxiolytic effects without cognitive defects. In contrast, we show that dual blockade of the endocannabinoid-degrading enzymes monoacylglycerol lipase (MAGL) and FAAH by selected organophosphorus agents leads to greater than ten-fold elevations in brain levels of both 2-AG and anandamide and to robust CB(1)-dependent behavioral effects that mirror those observed with CB(1) agonists. Arachidonic acid levels are decreased by the organophosphorus agents in amounts equivalent to elevations in 2-AG, which indicates that endocannabinoid and eicosanoid signaling pathways may be coordinately regulated in the brain.
ESTHER : Nomura_2008_Nat.Chem.Biol_4_373
PubMedSearch : Nomura_2008_Nat.Chem.Biol_4_373
PubMedID: 18438404

Title : Serine hydrolase KIAA1363: toxicological and structural features with emphasis on organophosphate interactions - Nomura_2006_Chem.Res.Toxicol_19_1142
Author(s) : Nomura DK , Durkin KA , Chiang KP , Quistad GB , Cravatt BF , Casida JE
Ref : Chemical Research in Toxicology , 19 :1142 , 2006
Abstract : Serine hydrolase KIAA1363 is highly expressed in invasive cancer cells and is the major protein in mouse brain diethylphosphorylated by and hydrolyzing low levels of chlorpyrifos oxon (CPO) (the activated metabolite of a major insecticide). It is also the primary CPO-hydrolyzing enzyme in spinal cord, kidney, heart, lung, testis, and muscle but not liver, a pattern of tissue expression confirmed by fluorophosphonate-rhodamine labeling. KIAA1363 gene deletion using homologous recombination reduces CPO binding, hydrolysis, and metabolism 3-29-fold on incubation with brain membranes and homogenates determined with 1 nM [(3)H-ethyl]CPO and the inhibitory potency for residual CPO with butyrylcholinesterase as a biomarker. Studies with knockout mice further show that KIAA1363 partially protects brain AChE and monoacylglycerol lipase from CPO-induced in vivo inhibition. Surprisingly, mouse brain KIAA1363 and AChE are similar in in vitro sensitivity to seven methyl, ethyl, and propyl but not higher alkyl OP insecticides and analogues, prompting structural comparisons of the active sites of KIAA1363 and AChE relative to OP potency and selectivity. Homology modeling based largely on the Archaeoglobus fulgidus esterase crystal structure indicates that KIAA1363 has a catalytic triad of S191, D348, and H378, a GDSAG motif, and an oxyanion hole of H113, G114, G115, and G116. Excellent selectivity for KIAA1363 is achieved on OP structure optimization with long alkyl chain substituents suggesting that KIAA1363 has larger acyl and leaving group pockets than those of AChE. KIAA1363 reactivates faster than AChE presumably due to differences in the uncoupling of the catalytic triad His upon phosphorylation. The structural modeling of KIAA1363 helps us understand OP structure-activity relationships and the toxicological relevance of this detoxifying enzyme.
ESTHER : Nomura_2006_Chem.Res.Toxicol_19_1142
PubMedSearch : Nomura_2006_Chem.Res.Toxicol_19_1142
PubMedID: 16978018
Gene_locus related to this paper: human-NCEH1 , mouse-Q8BLF1

Title : Each lipase has a unique sensitivity profile for organophosphorus inhibitors - Quistad_2006_Toxicol.Sci_91_166
Author(s) : Quistad GB , Liang SN , Fisher KJ , Nomura DK , Casida JE
Ref : Toxicol Sci , 91 :166 , 2006
Abstract : Lipases sensitive to organophosphorus (OP) inhibitors play critical roles in cell regulation, nutrition, and disease, but little is known on the toxicological aspects in mammals. To help fill this gap, six lipases or lipase-like proteins are assayed for OP sensitivity in vitro under standard conditions (25 degrees C, 15 min incubation). Postheparin serum lipase, lipoprotein lipase (LPL) (two sources), pancreatic lipase, monoacylglycerol (MAG) lipase, cholesterol esterase, and KIAA1363 are considered with 32 OP pesticides and related compounds. Postheparin lipolytic activity in rat serum is inhibited by 14 OPs, including chlorpyrifos oxon (IC50 50-97 nM). LPL (bovine milk and Pseudomonas) generally is less inhibited by the insecticides or activated oxons, but the milk enzyme is very sensitive to six fluorophosphonates and benzodioxaphosphorin oxides (IC50 7-20 nM). Porcine pancreatic lipase is very sensitive to dioctyl 4-nitrophenyl phosphate (IC50 8 nM), MAG lipase of mouse brain to O-4-nitrophenyl methyldodecylphosphinate (IC50 0.6 nM), and cholesterol esterase (bovine pancreas) to all of the classes of OPs tested (IC50 < 10 nM for 17 compounds). KIAA1363 is sensitive to numerous OPs, including two O-4-nitrophenyl compounds (IC50 3-4 nM). In an overview, inhibition of 28 serine hydrolases (including lipases) by eight OPs (chlorpyrifos oxon, diazoxon, paraoxon, dichlorvos, and four nonpesticides) showed that brain acetylcholinesterase is usually less sensitive than butyrylcholinesterase, liver esterase, cholesterol esterase, and KIAA1363. In general, each lipase (like each serine hydrolase) has a different spectrum of OP sensitivity, and individual OPs have unique ranking of potency for inhibition of serine hydrolases.
ESTHER : Quistad_2006_Toxicol.Sci_91_166
PubMedSearch : Quistad_2006_Toxicol.Sci_91_166
PubMedID: 16449251

Title : A brain detoxifying enzyme for organophosphorus nerve poisons - Nomura_2005_Proc.Natl.Acad.Sci.U.S.A_102_6195
Author(s) : Nomura DK , Leung D , Chiang KP , Quistad GB , Cravatt BF , Casida JE
Ref : Proc Natl Acad Sci U S A , 102 :6195 , 2005
Abstract : Organophosphorus (OP) insecticides and chemical warfare agents act primarily by inhibiting acetylcholinesterase. There are many secondary targets for OP toxicants as observed for example with the major insecticide chlorpyrifos and its bioactivated metabolite chlorpyrifos oxon (CPO). Therefore, it was surprising that the predominant mouse brain protein labeled in vitro by [(3)H-ethyl]CPO (1 nM) (designated CPO-binding protein or CPO-BP) is not one of these known OP toxicant targets. CPO-BP is a 50-kDa membrane-bound serine hydrolase measured by derivatization with [(3)H]CPO and SDS/PAGE or filtration binding assay. It appears to undergo rapid diethylphosphorylation by [(3)H]CPO followed by either dephosphorylation and reactivation or aging on loss of an ethyl group. CPO and several other OP toxicants potently inhibit CPO-BP in vivo (i.p., 2 h) (50% inhibition at 2-25 mg/kg) and in vitro (50% inhibition at 8-68 nM). Using three chemical labeling reagents, i.e., [(3)H]CPO and the activity-based proteomic probes fluorophosphonate-biotin and fluorophosphonate-rhodamine, mouse brain CPO-BP is identified as serine hydrolase KIAA1363 of unknown function. Brains from KIAA1363(-/-) mice show greatly reduced levels of CPO labeling and hydrolytic metabolism compared to brains from wild-type mice. KIAA1363 therefore is the principal enzyme for metabolizing low levels of CPO in brain and may play a more general role in detoxification of OP nerve poisons.
ESTHER : Nomura_2005_Proc.Natl.Acad.Sci.U.S.A_102_6195
PubMedSearch : Nomura_2005_Proc.Natl.Acad.Sci.U.S.A_102_6195
PubMedID: 15840715
Gene_locus related to this paper: human-NCEH1 , mouse-Q8BLF1

Title : Arachidonylsulfonyl derivatives as cannabinoid CB1 receptor and fatty acid amide hydrolase inhibitors - Segall_2003_Bioorg.Med.Chem.Lett_13_3301
Author(s) : Segall Y , Quistad GB , Nomura DK , Casida JE
Ref : Bioorganic & Medicinal Chemistry Lett , 13 :3301 , 2003
Abstract : Arachidonylsulfonyl fluoride (3), reported here for the first time, is similar in potency to its known methyl arachidonylfluorophosphonate (2) analogue as an inhibitor of mouse brain fatty acid amide hydrolase activity (IC(50) 0.1 nM) and cannabinoid CB1 agonist [3H]CP 55,940 binding (IC(50) 304-530 nM). Interestingly, 3 is much more selective than 2 as an inhibitor for fatty acid amide hydrolase relative to acetylcholinesterase, butyrylcholinesterase and neuropathy target esterase. N-(2-Hydroxyethyl)arachidonylsulfonamide (4) is at least 2500-fold less potent than N-(2-hydroxyethyl)arachidonamide (anandamide) (1) at the CB1 agonist site.
ESTHER : Segall_2003_Bioorg.Med.Chem.Lett_13_3301
PubMedSearch : Segall_2003_Bioorg.Med.Chem.Lett_13_3301
PubMedID: 12951114

Title : Toxicological and structural features of organophosphorus and organosulfur cannabinoid CB1 receptor ligands - Segall_2003_Toxicol.Sci_76_131
Author(s) : Segall Y , Quistad GB , Sparks SE , Nomura DK , Casida JE
Ref : Toxicol Sci , 76 :131 , 2003
Abstract : Potent cannabinoid CB1 receptor ligands include anandamide [N-(2-hydroxyethyl)arachidonamide], Delta9-tetrahydrocannabinol, and 3H-CP 55,940 at the agonist site and selected organophosphorus esters (including some pesticides) and organosulfur compounds at a proposed closely coupled "nucleophilic" site. This study considers the toxicological and structural features of alkylfluorophosphonates, benzodioxaphosphorin oxides, alkanesulfonyl fluorides, and analogs acting at the nucleophilic site. Binding at the agonist site, using3H-CP 55,940 in assays with mouse brain membranes, is inhibited byO-isopropyl dodecylfluorophosphonate (compound 2), dodecanesulfonyl fluoride (compound 14) and dodecylbenzodioxaphosphorin oxide with IC50 values of 2-11 nM. Compounds 2 and 14 are also effectivein vivo, with 84% inhibition of mouse brain CB1 binding 4 h after intraperitoneal dosage at 30 mg/kg. Compound 14-inhibited CB1 in mouse brain requires about 3-4 days for recovery of 50% activity, suggesting covalent derivatization. Delayed toxicity (mortality in 0.3-5 days) from compounds 2, 14, and octanesulfonyl fluoride (18) is more closely associated with in vivo inhibition of brain neuropathy target esterase-lysophospholipase (NTE-LysoPLA) than with that of CB1 or acetylcholinesterase. NTE-LysoPLA inhibited by sulfonyl fluorides 14 and 18 cannot "age," a proposed requirement for NTE phosphorylated by organophosphorus-delayed neurotoxicants. Several octane- and dodecanesulfonamides with N-(2-hydroxyethyl) and other substituents based on anandamide give depressed mobility and recumbent posture in mice, but the effects do not correlate with potency for CB1 inhibition in vitro. Specific toxicological responses are not clearly associated with organophosphorus- or organosulfur-induced inhibition of the proposed CB1 nucleophilic site in mouse brain. On the other hand, the most potent CB1 inhibitors examined here are also NTE-LysoPLA inhibitors and cause delayed toxicity in mice.
ESTHER : Segall_2003_Toxicol.Sci_76_131
PubMedSearch : Segall_2003_Toxicol.Sci_76_131
PubMedID: 12944586

Title : Selective inhibitors of fatty acid amide hydrolase relative to neuropathy target esterase and acetylcholinesterase: toxicological implications - Quistad_2002_Toxicol.Appl.Pharmacol_179_57
Author(s) : Quistad GB , Sparks SE , Segall Y , Nomura DK , Casida JE
Ref : Toxicol Appl Pharmacol , 179 :57 , 2002
Abstract : Fatty acid amide hydrolase (FAAH) plays an important role in nerve function by regulating the action of endocannabinoids (e.g., anandamide) and hydrolyzing a sleep-inducing factor (oleamide). Several organophosphorus pesticides and related compounds are shown in this study to be more potent in vivo inhibitors of mouse brain FAAH than neuropathy target esterase (NTE), raising the question of the potential toxicological relevance of FAAH inhibition. These FAAH-selective compounds include tribufos and (R)-octylbenzodioxaphosphorin oxide with delayed neurotoxic effects in mice and hens plus several organophosphorus pesticides (e.g., fenthion) implicated as delayed neurotoxicants in humans. The search for a highly potent and selective inhibitor for FAAH relative to NTE for use as a toxicological probe culminated in the discovery that octylsulfonyl fluoride inhibits FAAH by 50% at 2 nM in vitro and 0.2 mg/kg in vivo and NTE is at least 100-fold less sensitive in each case. More generally, the studies revealed 12 selective in vitro inhibitors for FAAH (mostly octylsulfonyl and octylphosphonyl derivatives) and 9 for NTE (mostly benzodioxaphosphorin oxides and organophosphorus fluoridates). The overall in vivo findings with 16 compounds indicate the expected association of AChE inhibition with acute or cholinergic syndrome and >70% brain NTE inhibition with delayed neurotoxic action. Surprisingly, 75-99% brain FAAH inhibition does not lead to any overt neurotoxicity or change in behavior (other than potentiation of exogenous anandamide action). Thus, FAAH inhibition in mouse brain does not appear to be a primary target for organophosphorus pesticide-induced neurotoxic action (cholinergic or intermediate syndrome or delayed neurotoxicity).
ESTHER : Quistad_2002_Toxicol.Appl.Pharmacol_179_57
PubMedSearch : Quistad_2002_Toxicol.Appl.Pharmacol_179_57
PubMedID: 11884237

Title : Cannabinoid CB1 receptor as a target for chlorpyrifos oxon and other organophosphorus pesticides - Quistad_2002_Toxicol.Lett_135_89
Author(s) : Quistad GB , Nomura DK , Sparks SE , Segall Y , Casida JE
Ref : Toxicol Lett , 135 :89 , 2002
Abstract : Binding of the endocannabinoid anandamide or of Delta(9)-tetrahydrocannabinol to the agonist site of the cannabinoid receptor (CB1) is commonly assayed with [3H]CP 55,940. Potent long-chain alkylfluorophosphonate inhibitors of agonist binding suggest an additional, important and closely-coupled nucleophilic site, possibly undergoing phosphorylation. We find that the CB1 receptor is also sensitive to inhibition in vitro and in vivo by several organophosphorus pesticides and analogs. Binding of [3H]CP 55,940 to mouse brain CB1 receptor in vitro is inhibited 50% by chlorpyrifos oxon at 14 nM, chlorpyrifos methyl oxon at 64 nM and paraoxon, diazoxon and dichlorvos at 1200-4200 nM. Some 15 other organophosphorus pesticides and analogs are less active in vitro. The plant defoliant tribufos inhibits CB1 in vivo, without cholinergic poisoning signs, by 50% at 50 mg/kg intraperitoneally with a recovery half-time of 3-4 days, indicating covalent derivatization. [3H-ethyl]Chlorpyrifos oxon may be suitable for radiolabeling and characterization of this proposed nucleophilic site.
ESTHER : Quistad_2002_Toxicol.Lett_135_89
PubMedSearch : Quistad_2002_Toxicol.Lett_135_89
PubMedID: 12243867