Stella N

References (12)

Title : P2X(7) receptor-dependent increase in endocannabinoid 2-arachidonoyl glycerol production by neuronal cells in culture: Dynamics and mechanism - Singh_2024_Br.J.Pharmacol__
Author(s) : Singh S , Sarroza D , English A , Whittington D , Dong A , Malamas M , Makriyannis A , van der Stelt M , Li Y , Zweifel L , Bruchas MR , Land BB , Stella N
Ref : British Journal of Pharmacology , : , 2024
Abstract : BACKGROUND AND PURPOSE: Neurotransmission and neuroinflammation are controlled by local increases in both extracellular ATP and the endocannabinoid 2-arachidonoyl glycerol (2-AG). While it is known that extracellular ATP stimulates 2-AG production in cells in culture, the dynamics and molecular mechanisms that underlie this response remain poorly understood. Detection of real-time changes in eCB levels with the genetically encoded sensor, GRAB(eCB2.0), can address this shortfall. EXPERIMENTAL APPROACH: 2-AG and arachidonoylethanolamide (AEA) levels in Neuro2a (N2a) cells were measured by LC-MS, and GRAB(eCB2.0) fluorescence changes were detected using live-cell confocal microscopy and a 96-well fluorescence plate reader. KEY RESULTS: 2-AG and AEA increased GRAB(eCB2.0) fluorescence in N2a cells with EC(50) values of 81 and 58 nM, respectively; both responses were reduced by the cannabinoid receptor type 1 (CB(1)R) antagonist SR141617 and absent in cells expressing the mutant-GRAB(eCB2.0). ATP increased only 2-AG levels in N2a cells, as measured by LC-MS, and induced a transient increase in the GRAB(eCB2.0) signal within minutes primarily via activation of P2X(7) receptors (P2X(7)R). This response was dependent on diacylglycerol lipase beta activity, partially dependent on extracellular calcium and phospholipase C activity, but not controlled by the 2-AG hydrolysing enzyme, alpha/beta-hydrolase domain containing 6 (ABHD6). CONCLUSIONS AND IMPLICATIONS: Considering that P2X(7)R activation increases 2-AG levels within minutes, our results show how these molecular components are mechanistically linked. The specific molecular components in these signalling systems represent potential therapeutic targets for the treatment of neurological diseases, such as chronic pain, that involve dysregulated neurotransmission and neuroinflammation.
ESTHER : Singh_2024_Br.J.Pharmacol__
PubMedSearch : Singh_2024_Br.J.Pharmacol__
PubMedID: 38581262

Title : ABHD6 and MAGL control 2-AG levels in the PAG and allodynia in a CSD-induced periorbital model of headache - Liktor-Busa_2023_Front.Pain.Res.(Lausanne)_4_1171188
Author(s) : Liktor-Busa E , Levine AA , Palomino SM , Singh S , Wahl J , Vanderah TW , Stella N , Largent-Milnes TM
Ref : Front Pain Res (Lausanne) , 4 :1171188 , 2023
Abstract : INTRODUCTION: The high prevalence and severe symptoms of migraines in humans emphasizes the need to identify underlying mechanisms that can be targeted for therapeutic benefit. Clinical Endocannabinoid Deficiency (CED) posits that reduced endocannabinoid tone may contribute to migraine development and other neuropathic pain conditions. While strategies that increase levels of the endocannabinoid n-arachidonoylethanolamide have been tested, few studies have investigated targeting the levels of the more abundant endocannabinoid, 2-arachidonoylgycerol, as an effective migraine intervention. METHODS: Cortical spreading depression was induced in female Sprague Dawley rats via KCl (potassium chloride) administration, followed by measures of endocannabinoid levels, enzyme activity, and neuroinflammatory markers. Efficacy of inhibiting 2-arachidonoylglycerol hydrolysis to mitigate periorbital allodynia was then tested using reversal and prevention paradigms. RESULTS: We discovered reduced 2-arachidonoylglycerol levels in the periaqueductal grey associated with increased hydrolysis following headache induction. Pharmacological inhibition of the 2-arachidonoylglycerol hydrolyzing enzymes, alpha/beta-hydrolase domain-containing 6 and monoacylglycerol lipase reversed and prevented induced periorbital allodynia in a cannabinoid receptor-dependent manner. DISCUSSION: Our study unravels a mechanistic link between 2-arachidonoylglycerol hydrolysis activity in the periaqueductal grey in a preclinical, rat model of migraine. Thus, 2-arachidonoylglycerol hydrolysis inhibitors represent a potential new therapeutic avenue for the treatment of headache.
ESTHER : Liktor-Busa_2023_Front.Pain.Res.(Lausanne)_4_1171188
PubMedSearch : Liktor-Busa_2023_Front.Pain.Res.(Lausanne)_4_1171188
PubMedID: 37287623
Gene_locus related to this paper: human-ABHD6 , human-MGLL

Title : The serine hydrolase ABHD6 controls survival and thermally induced seizures in a mouse model of Dravet syndrome - Westenbroek_2023_Neurobiol.Dis_180_106099
Author(s) : Westenbroek R , Kaplan J , Viray K , Stella N
Ref : Neurobiol Dis , 180 :106099 , 2023
Abstract : Evidence suggests that inhibition of alpha/beta hydrolase-domain containing 6 (ABHD6) reduces seizures; however, the molecular mechanism of this therapeutic response remains unknown. We discovered that heterozygous expression of Abhd6 (Abhd6(+/-)) significantly reduced the premature lethality of Scn1a(+/-) mouse pups, a genetic mouse model of Dravet Syndrome (DS). Both Abhd6(+/-) mutation and pharmacological inhibition of ABHD6 reduced the duration and incidence of thermally induced seizures in Scn1a(+/-) pups. Mechanistically, the in vivo anti-seizure response resulting from ABHD6 inhibition is mediated by potentiation of gamma-aminobutyric acid receptors Type-A (GABA(A)R). Brain slice electrophysiology showed that blocking ABHD6 potentiates extrasynaptic (tonic) GABA(A)R currents that reduce dentate granule cell excitatory output without affecting synaptic (phasic) GABA(A)R currents. Our results unravel an unexpected mechanistic link between ABHD6 activity and extrasynaptic GABA(A)R currents that controls hippocampal hyperexcitability in a genetic mouse model of DS. BRIEF SUMMARY: This study provides the first evidence for a mechanistic link between ABHD6 activity and the control of extrasynaptic GABA(A)R currents that controls hippocampal hyperexcitability in a genetic mouse model of Dravet Syndrome and can be targeted to dampened seizures.
ESTHER : Westenbroek_2023_Neurobiol.Dis_180_106099
PubMedSearch : Westenbroek_2023_Neurobiol.Dis_180_106099
PubMedID: 36990366
Gene_locus related to this paper: human-ABHD6 , mouse-ABHD6

Title : ABHD6 Controls Amphetamine-Stimulated Hyperlocomotion: Involvement of CB(1) Receptors - Deng_2021_Cannabis.Cannabinoid.Res__
Author(s) : Deng L , Viray K , Singh S , Cravatt B , Stella N
Ref : Cannabis Cannabinoid Res , : , 2021
Abstract : Introduction: Activation of cannabinoid 1 receptors (CB(1)Rs) by endocannabinoids (eCBs) is controlled by both eCB production and eCB inactivation. Accordingly, inhibition of eCB hydrolyzing enzymes, monoacylglycerol lipase (MAGL) and alpha/beta-hydrolase domain containing 6 (ABHD6), enhances eCB accumulation and CB(1)R activation. It is known that inhibition of MAGL regulates select CB(1)R-dependent behaviors in mice, including locomotor behaviors and their modulation by psychostimulants, but much less is known about the effect of inhibiting ABHD6 activity on such behaviors. Methods: We report a new mouse line that carries a genetic deletion of Abhd6 and evaluated its effect on spontaneous locomotion measured in a home cage monitoring system, motor coordination measured on a Rotarod, and amphetamine-stimulated hyperlocomotion and amphetamine sensitization (AS) measured in an open-field chamber. Results: ABHD6 knockout (KO) mice reached adulthood without exhibiting overt behavioral impairment, and we measured only mild reduction in spontaneous locomotion and motor coordination in adult ABHD6 KO mice compared to wild-type (WT) mice. Significantly, amphetamine-stimulated hyperlocomotion was enhanced by twofold in ABHD6 KO mice compared to WT mice and yet ABHD6 KO mice expressed AS to the same extent as WT mice. A twofold increase in amphetamine-stimulated hyperlocomotion was also measured in ABHD6 heterozygote mice and in WT mice treated with the ABHD6 inhibitor KT-182. It is known that amphetamine-stimulated hyperlocomotion is not affected by the CB(1)R antagonist, SR141617, and we discovered that the enhanced amphetamine-stimulated hyperlocomotion resulting from ABHD6 inhibition is blocked by SR141617. Conclusions: Our study suggests that ABHD6 controls amphetamine-stimulated hyperlocomotion by a mechanistic switch to a CB(1)R-dependent mechanism.
ESTHER : Deng_2021_Cannabis.Cannabinoid.Res__
PubMedSearch : Deng_2021_Cannabis.Cannabinoid.Res__
PubMedID: 34705543
Gene_locus related to this paper: human-ABHD6 , mouse-ABHD6

Title : Benzisothiazolinone derivatives as potent allosteric monoacylglycerol lipase inhibitors that functionally mimic sulfenylation of regulatory cysteines - Castelli_2020_J.Med.Chem_63_1261
Author(s) : Castelli R , Scalvini L , Vacondio F , Lodola A , Anselmi M , Vezzosi S , Carmi C , Bassi M , Ferlenghi F , Rivara S , Moller IR , Rand KD , Daglian J , Wei D , Dotsey EY , Ahmed F , Jung KM , Stella N , Singh S , Mor M , Piomelli D
Ref : Journal of Medicinal Chemistry , 63 :1261 , 2020
Abstract : We describe a set of benzisothiazolinone (BTZ) derivatives that are potent inhibitors of monoacylglycerol lipase (MGL), the primary degrading enzyme for the endocannabinoid 2-arachidonoyl-sn-glycerol (2-AG). Structure-activity relationship studies evaluated various substitutions on the nitrogen atom and the benzene ring of the BTZ nucleus. Optimized derivatives with nanomolar potency allowed to investigate the mechanism of MGL inhibition. Site-directed mutagenesis and mass spectrometry experiments showed that BTZs interact in a covalent reversible manner with regulatory cysteines, Cys201 and Cys208, causing a reversible sulfenylation known to modulate MGL activity. Metadynamics simulations revealed that BTZ adducts favor a closed conformation of MGL that occludes substrate recruitment. The BTZ derivative 13 protected neuronal cells from oxidative stimuli and increased 2-AG levels in mouse brain. The results identify Cys201 and Cys208 as key regulators of MGL function, and point to the BTZ scaffold as a useful starting point for the discovery of allosteric MGL inhibitors.
ESTHER : Castelli_2020_J.Med.Chem_63_1261
PubMedSearch : Castelli_2020_J.Med.Chem_63_1261
PubMedID: 31714779

Title : Identification of alpha,beta-Hydrolase Domain Containing Protein 6 as a Diacylglycerol Lipase in Neuro-2a Cells - van Esbroeck_2019_Front.Mol.Neurosci_12_286
Author(s) : van Esbroeck ACM , Kantae V , Di X , van der Wel T , den Dulk H , Stevens AF , Singh S , Bakker AT , Florea BI , Stella N , Overkleeft HS , Hankemeier T , van der Stelt M
Ref : Front Mol Neurosci , 12 :286 , 2019
Abstract : The endocannabinoid 2-arachidonoylglycerol (2-AG) is involved in neuronal differentiation. This study aimed to identify the biosynthetic enzymes responsible for 2-AG production during retinoic acid (RA)-induced neurite outgrowth of Neuro-2a cells. First, we confirmed that RA stimulation of Neuro-2a cells increases 2-AG production and neurite outgrowth. The diacylglycerol lipase (DAGL) inhibitor DH376 blocked 2-AG production and reduced neuronal differentiation. Surprisingly, CRISPR/Cas9-mediated knockdown of DAGLalpha and DAGLbeta in Neuro-2a cells did not reduce 2-AG levels, suggesting another enzyme capable of producing 2-AG in this cell line. Chemical proteomics revealed DAGLbeta and alpha,beta-hydrolase domain containing protein (ABHD6) as the only targets of DH376 in Neuro-2a cells. Biochemical, genetic and lipidomic studies demonstrated that ABHD6 possesses DAGL activity in conjunction with its previously reported monoacylglycerol lipase activity. RA treatment of Neuro-2a cells increased by three-fold the amount of active ABHD6. Our study shows that ABHD6 exhibits significant DAG lipase activity in Neuro-2a cells in addition to its known MAG lipase activity and suggest it is involved in neuronal differentiation.
ESTHER : van Esbroeck_2019_Front.Mol.Neurosci_12_286
PubMedSearch : van Esbroeck_2019_Front.Mol.Neurosci_12_286
PubMedID: 31849602
Gene_locus related to this paper: human-ABHD6

Title : ABHD6: Its Place in Endocannabinoid Signaling and Beyond - Cao_2019_Trends.Pharmacol.Sci_40_267
Author(s) : Cao JK , Kaplan J , Stella N
Ref : Trends in Pharmacological Sciences , 40 :267 , 2019
Abstract : The endocannabinoid (eCB) signaling system modulates neurotransmission and inflammation, among other physiological functions. Its newest member, alpha/beta-hydrolase domain-containing 6 (ABHD6), has emerged as a promising therapeutic target to treat several devastating diseases, including epilepsy. Here, we review the molecular mechanisms that mediate and control eCB signaling and, within it, the specific role of ABHD6. We also discuss how ABHD6 controls the abundance of additional lipids and the trafficking of ionotropic receptors to plasma membranes. We finish with several unexplored questions regarding this novel enzyme. Our current understanding of the molecular mechanism and biological function of ABHD6 provides a strong foundation for the development of small-molecule therapeutics to treat devastating diseases.
ESTHER : Cao_2019_Trends.Pharmacol.Sci_40_267
PubMedSearch : Cao_2019_Trends.Pharmacol.Sci_40_267
PubMedID: 30853109
Gene_locus related to this paper: human-ABHD6 , mouse-ABHD6

Title : ABHD6 Blockade Exerts Antiepileptic Activity in PTZ-Induced Seizures and in Spontaneous Seizures in R6\/2 Mice - Naydenov_2014_Neuron_83_361
Author(s) : Naydenov AV , Horne EA , Cheah CS , Swinney K , Hsu KL , Cao JK , Marrs WR , Blankman JL , Tu S , Cherry AE , Fung S , Wen A , Li W , Saporito MS , Selley DE , Cravatt BF , Oakley JC , Stella N
Ref : Neuron , 83 :361 , 2014
Abstract : The serine hydrolase alpha/beta-hydrolase domain 6 (ABHD6) hydrolyzes the most abundant endocannabinoid (eCB) in the brain, 2-arachidonoylglycerol (2-AG), and controls its availability at cannabinoid receptors. We show that ABHD6 inhibition decreases pentylenetetrazole (PTZ)-induced generalized tonic-clonic and myoclonic seizure incidence and severity. This effect is retained in Cnr1(-/-) or Cnr2(-/-) mice, but blocked by addition of a subconvulsive dose of picrotoxin, suggesting the involvement of GABAA receptors. ABHD6 inhibition also blocked spontaneous seizures in R6/2 mice, a genetic model of juvenile Huntington's disease known to exhibit dysregulated eCB signaling. ABHD6 blockade retained its antiepileptic activity over chronic dosing and was not associated with psychomotor or cognitive effects. While the etiology of seizures in R6/2 mice remains unsolved, involvement of the hippocampus is suggested by interictal epileptic discharges, increased expression of vGLUT1 but not vGAT, and reduced Neuropeptide Y (NPY) expression. We conclude that ABHD6 inhibition may represent a novel antiepileptic strategy.
ESTHER : Naydenov_2014_Neuron_83_361
PubMedSearch : Naydenov_2014_Neuron_83_361
PubMedID: 25033180
Gene_locus related to this paper: human-ABHD6 , mouse-ABHD6

Title : Two Novel Mutations in ABHD12: Expansion of the Mutation Spectrum in PHARC and Assessment of Their Functional Effects - Chen_2013_Hum.Mutat_34_1672
Author(s) : Chen DH , Naydenov A , Blankman JL , Mefford HC , Davis M , Sul Y , Barloon AS , Bonkowski E , Wolff J , Matsushita M , Smith C , Cravatt BF , Mackie K , Raskind WH , Stella N , Bird TD
Ref : Hum Mutat , 34 :1672 , 2013
Abstract : PHARC (polyneuropathy, hearing loss, ataxia, retinitis pigmentosa, and cataracts) is a recently described autosomal-recessive neurodegenerative disease caused by mutations in the alpha-beta-hydrolase domain-containing 12 gene (ABHD12). Only five homozygous ABHD12 mutations have been reported and the pathogenesis of PHARC remains unclear. We evaluated a woman who manifested short stature as well as the typical features of PHARC. Sequence analysis of ABHD12 revealed a novel heterozygous c.1129A>T (p.Lys377*) mutation. Targeted comparative genomic hybridization detected a 59-kb deletion that encompasses exon 1 of ABHD12 and exons 1-4 of an adjacent gene, GINS1, and includes the promoters of both genes. The heterozygous deletion was also carried by the patient's asymptomatic mother. Quantitative reverse transcription-PCR demonstrated approximately 50% decreased expression of ABHD12 RNA in lymphoblastoid cell lines from both individuals. Activity-based protein profiling of serine hydrolases revealed absence of ABHD12 hydrolase activity in the patient and 50% reduction in her mother. This is the first report of compound heterozygosity in PHARC and the first study to describe how a mutation might affect ABHD12 expression and function. The possible involvement of haploinsufficiency for GINS1, a DNA replication complex protein, in the short stature of the patient and her mother requires further studies.
ESTHER : Chen_2013_Hum.Mutat_34_1672
PubMedSearch : Chen_2013_Hum.Mutat_34_1672
PubMedID: 24027063
Gene_locus related to this paper: human-ABHD12

Title : Dual inhibition of alpha\/beta-hydrolase domain 6 and fatty acid amide hydrolase increases endocannabinoid levels in neurons - Marrs_2011_J.Biol.Chem_286_28723
Author(s) : Marrs WR , Horne EA , Ortega-Gutierrez S , Cisneros JA , Xu C , Lin YH , Muccioli GG , Lopez-Rodriguez ML , Stella N
Ref : Journal of Biological Chemistry , 286 :28723 , 2011
Abstract : Agonists at cannabinoid receptors, such as the phytocannabinoid Delta(9)-tetrahydrocannabinol, exert a remarkable array of therapeutic effects but are also associated with undesirable psychoactive side effects. Conversely, targeting enzymes that hydrolyze endocannabinoids (eCBs) allows for more precise fine-tuning of cannabinoid receptor signaling, thus providing therapeutic relief with reduced side effects. Here, we report the development and characterization of an inhibitor of eCB hydrolysis, UCM710, which augments both N-arachidonoylethanolamine and 2-arachidonoylglycerol levels in neurons. This compound displays a unique pharmacological profile in that it inhibits fatty acid amide hydrolase and alpha/beta-hydrolase domain 6 but not monoacylglycerol lipase. Thus, UCM710 represents a novel tool to delineate the therapeutic potential of compounds that manipulate a subset of enzymes that control eCB signaling.
ESTHER : Marrs_2011_J.Biol.Chem_286_28723
PubMedSearch : Marrs_2011_J.Biol.Chem_286_28723
PubMedID: 21665953
Gene_locus related to this paper: human-ABHD6

Title : The serine hydrolase ABHD6 controls the accumulation and efficacy of 2-AG at cannabinoid receptors - Marrs_2010_Nat.Neurosci_13_951
Author(s) : Marrs WR , Blankman JL , Horne EA , Thomazeau A , Lin YH , Coy J , Bodor AL , Muccioli GG , Hu SS , Woodruff G , Fung S , Lafourcade M , Alexander JP , Long JZ , Li W , Xu C , Moller T , Mackie K , Manzoni OJ , Cravatt BF , Stella N
Ref : Nat Neurosci , 13 :951 , 2010
Abstract : The endocannabinoid 2-arachidonoylglycerol (2-AG) regulates neurotransmission and neuroinflammation by activating CB1 cannabinoid receptors on neurons and CB2 cannabinoid receptors on microglia. Enzymes that hydrolyze 2-AG, such as monoacylglycerol lipase, regulate the accumulation and efficacy of 2-AG at cannabinoid receptors. We found that the recently described serine hydrolase alpha-beta-hydrolase domain 6 (ABHD6) also controls the accumulation and efficacy of 2-AG at cannabinoid receptors. In cells from the BV-2 microglia cell line, ABHD6 knockdown reduced hydrolysis of 2-AG and increased the efficacy with which 2-AG can stimulate CB2-mediated cell migration. ABHD6 was expressed by neurons in primary culture and its inhibition led to activity-dependent accumulation of 2-AG. In adult mouse cortex, ABHD6 was located postsynaptically and its selective inhibition allowed the induction of CB1-dependent long-term depression by otherwise subthreshold stimulation. Our results indicate that ABHD6 is a rate-limiting step of 2-AG signaling and is therefore a bona fide member of the endocannabinoid signaling system.
ESTHER : Marrs_2010_Nat.Neurosci_13_951
PubMedSearch : Marrs_2010_Nat.Neurosci_13_951
PubMedID: 20657592
Gene_locus related to this paper: human-ABHD6

Title : Identification of a novel endocannabinoid-hydrolyzing enzyme expressed by microglial cells - Muccioli_2007_J.Neurosci_27_2883
Author(s) : Muccioli GG , Xu C , Odah E , Cudaback E , Cisneros JA , Lambert DM , Lopez Rodriguez ML , Bajjalieh S , Stella N
Ref : Journal of Neuroscience , 27 :2883 , 2007
Abstract : The endocannabinoids (eCBs) anandamide and 2-arachidonoyl glycerol (2-AG) are inactivated by a two-step mechanism. First, they are carried into cells, and then anandamide is hydrolyzed by fatty acid amide hydrolase (FAAH) and 2-AG by monoacylglycerol lipase (MGL). Here we provide evidence for a previously undescribed MGL activity expressed by microglial cells. We found that the mouse microglial cell line BV-2 does not express MGL mRNA and yet efficiently hydrolyzes 2-AG. URB597 (3'-carbamoyl-biphenyl-3-yl-cyclohexylcarbamate) reduces this hydrolysis by 50%, suggesting the involvement of FAAH. The remaining activity is blocked by classic MGL inhibitors [[1,1-biphenyl]-3-yl-carbamic acid, cyclohexyl ester (URB602) and MAFP (methylarachidonyl fluorophosphate)] and is unaffected by inhibitors of COXs (cyclooxygenases), LOXs (lipooxygenases), and DGLs (diacylglycerol lipases), indicating the involvement of a novel MGL activity. Accordingly, URB602 leads to selective accumulation of 2-AG in intact BV-2 cells. Although MGL expressed in neurons is equally distributed between the cytosolic, mitochondrial, and nuclear fractions, the novel MGL activity expressed by BV-2 cells is enriched in mitochondrial and nuclear fractions. A screen for novel inhibitors of eCB hydrolysis identified several compounds that differentially block MGL, FAAH, and the novel MGL activity. Finally, we provide evidence for expression of the novel MGL by mouse primary microglia in culture. Our results suggest the presence of a novel, pharmacologically distinct, MGL activity that controls 2-AG levels in microglia.
ESTHER : Muccioli_2007_J.Neurosci_27_2883
PubMedSearch : Muccioli_2007_J.Neurosci_27_2883
PubMedID: 17360910