OPRM1 Research Articles

Machine-learned analysis of global and glial/opioid intersection\u2013related DNA methylation in patients with persistent pain after breast cancer surgery

BackgroundGlial cells in the central nervous system play a key role in neuroinflammation and subsequent central sensitization to pain. They are therefore involved in the development of persistent pain. One of the main sites of interaction of the immune system with persistent pain has been identified as neuro-immune crosstalk at the glial-opioid interface. The present study examined a potential association between the DNA methylation of two key players of glial/opioid intersection and persistent postoperative pain.MethodsIn a cohort of 140 women who had undergone breast cancer surgery, and were assigned based on a 3-year follow-up to either a persistent or non-persistent pain phenotype, the role of epigenetic regulation of key players in the glial-opioid interface was assessed. The methylation of genes coding for the Toll-like receptor 4 (TLR4) as a major mediator of glial contributions to persistent pain or for the μ-opioid receptor (OPRM1) was analyzed and its association with the pain phenotype was compared with that conferred by global genome-wide DNA methylation assessed via quantification of the methylation in the retrotransposon LINE1.ResultsTraining of machine learning algorithms indicated that the global DNA methylation provided a similar diagnostic accuracy for persistent pain as previously established non-genetic predictors. However, the diagnosis can be based on a single DNA based marker. By contrast, the methylation of TLR4 or OPRM1 genes could not contribute further to the allocation of the patients to the pain-related phenotype groups.ConclusionsWhile clearly supporting a predictive utility of epigenetic testing, the present analysis cannot provide support for specific epigenetic modulation of persistent postoperative pain via methylation of two key genes of the glial-opioid interface.

Murine model of OPRM1 A118G alters oxycodone self-administration and locomotor activation, but not conditioned place preference

Mu-opioid receptors (MORs) mediate the rewarding properties of oxycodone and other prescription opioid medications, which have played a central role in the current opioid epidemic in the United States. The human mu-opioid receptor gene (OPRM1) contains a functional single nucleotide polymorphism (SNP), A118G, which has been associated with altered opioid addiction risk, however the mechanisms responsible for this are not well understood. To explore this, we examined oxycodone conditioned place preference (CPP) and self-administration behavior (SA) in A112G mice, which possess a functionally analogous SNP in the mouse mu-opioid receptor gene (Oprm1). For CPP, male and female A112G mice homozygous for the A112 (wild-type; AA) or G112 (GG) allele were conditioned with doses of 1 and 3 mg/kg across an 8-day period. For SA, mice were allowed to self administer oxycodone (unit dose 0.25 mg/kg/infusion, FR1) for 4h/day for 10 consecutive days. We observed no effects of genotype or sex on conditioned place preference behavior. Oxycodone 3 mg/kg increased locomotor activity in AA mice but not GG mice, and both male and female GG mice self-administered significantly more oxycodone compared to their wild-type AA littermates. These studies suggest that the G allele promotes greater opioid intake, which may underlie greater opioid addiction morbidity in G-allele carriers.

Correlation between gene polymorphism and opioid efficacy in patients with gastric or intestinal cancer.

To explore the correlation between gene polymorphism and opioid efficacy in patients with gastric or intestinal cancer. Fifty-nine patients who underwent laparoscopic surgery for gastric or intestinal cancer under general anesthesia were included and randomly divided into oxycodone (n=30) and sufentanil groups (n=29) by reproducible random number generation method. Single nucleotide polymorphisms (SNPs) of four alleles: μ-opioid receptor gene OPRM1 A118G, cytochrome P450 (CPY450) enzyme system: CPY3A4*1G, CYP3A5*3, and CYP2D6*10 were detected by PCR-pyrosequencing. Patients in sufentanil group received intravenous sufentanil injection during anesthesia induction, intraoperative maintenance, and postoperative analgesia, while those in oxycodone group received oxycodone. Patients' postoperative VAS score, opioid use, and prevalence of adverse reactions were recorded. The genotype distribution of OPRM1 A118G, CYP3A4*1G, CYP3A5*3, and CYP2D6*10 in Chinese gastric cancer/intestinal cancer patients accorded with the Hardy-Weinberg law (p>0.05). OPRM1 A118G polymorphism correlated with postoperative VAS score and medication dosage, in oxycodone group (p<0.05), while it didn't with those of sufentanil group. The VAS scores in GG group were higher than that in AA group and AG group at T6-T9, (p<0.05); the postoperative pain remedies times in GG group were more than that in the AA and AG groups (p=0.002). CYP3A4*1G polymorphism related to postoperative VAS score, medication dosage and prevalence of adverse reactions in sufentanil group (p<0.05), while it didn't with those of oxycodone group (p>0.05). The total intraoperative medication in AA group was less than that in GG and GA groups (p<0.01), with a higher prevalence of respiratory depression (p=0.01). Nor was there any correlation of CYP3A5*3 and CYP2D6*10 polymorphisms with the efficacy, postoperative VAS score, pain remedies times, postoperative 24 h medication dosage, or prevalence of adverse reactions in oxycodone and sufentanil groups. Gene polymorphism affects the efficacy and adverse reactions of opioids in patients undergoing laparoscopic gastric or intestinal cancer surgery.

Generation of a MOR-CreER knock-in mouse line to study cells and neural circuits involved in mu opioid receptor signaling.

Mu opioid receptor (MOR) is involved in various brain functions, such as pain modulation, reward processing, and addictive behaviors, and mediates the main pharmacologic effects of morphine and other opioid compounds. To gain genetic access to MOR-expressing cells, and to study physiological and pathological roles of MOR signaling, we generated a MOR-CreER knock-in mouse line, in which the stop codon of the Oprm1 gene was replaced by a DNA fragment encoding a T2A peptide and tamoxifen (Tm)-inducible Cre recombinase. We show that the MOR-CreER allele undergoes Tm-dependent recombination in a discrete subtype of neurons that express MOR in the adult nervous system, including the olfactory bulb, cerebral cortex, striosome compartments in the striatum, hippocampus, amygdala, thalamus, hypothalamus, interpeduncular nucleus, superior and inferior colliculi, periaqueductal gray, parabrachial nuclei, cochlear nucleus, raphe nuclei, pontine and medullary reticular formation, ambiguus nucleus, solitary nucleus, spinal cord, and dorsal root ganglia. The MOR-CreER mouse line combined with a Cre-dependent adeno-associated virus vector enables robust gene manipulation in the MOR-enriched striosomes. Furthermore, Tm treatment during prenatal development effectively induces Cre-mediated recombination. Thus, the MOR-CreER mouse is a powerful tool to study MOR-expressing cells with conditional gene manipulation in developing and mature neural tissues.

Polymorphism of μ1-opioid receptor (OPRM1) gene may be of value in genesis of renal malignant neoplasm

Objective: to evaluate alleles distribution of single nucleotide polymorphism 118A&gt;G of μ1-opioid receptor (OPRM1) gene in patients with renal neoplasm and benign diseases.Materials and methods. 100 consecutive patients after renal surgeries retrospectively divided into groups with neoplasm (n = 29) and benign diseases (n = 71).Results. The incidence of renal neoplasm was much higher in homozygous 118A patients than in AG + GG group (36.4 % vs. 14.7 %; p = 0.035).Conclusion. Single nucleotide polymorphism 118A&gt;G OPRM1 gene may be of value in genesis of renal neoplasm.

Comparing Oprm1 Gene Promoter Methylation in the Lymphocytes of Male Rats Addicted to Nicotine, Morphine, Methadone, and Buprenorphine

Background: Addiction is a polygenic disorder caused by genetic and environmental factors. The opioid material can act as an epigenetic element, like DNA methylation. Objectives: The present study aimed to examine the effect of epigenetic drugs such as nicotine, morphine, methadone, and buprenorphine on the methylation of two CpG sites in promoter of Oprm1 gene in male Wistar rats. Materials &amp; Methods: In this case-control study, 48 male Wistar rats with Mean±SD (200±30) g were divided into 6 groups: These are five groups only The control (intact) group, nicotine (0.4 mg/kg, SC injection for 5 days), morphine (10 mg/kg on days 1-3 and 20 mg/kg on days 4-6 and 40 mg/kg, IP injection on days 7-9), methadone (0.5 mg/kg, IP injection for 15 days), buprenorphine (0.05 mg/kg, SC injection for 6 days) and finally saline for each respected group. After the treatment, genomic DNA was extracted from the whole blood of the rats. Then, the extracted DNA was treated with sodium bisulfate. To identify the selected methylated areas of Oprm1 promoter sites (CpG-107 and CpG+33), we used methylation-specific PCR (MSP). Results: Our results showed no methylation in the two CpG sites of Oprm1 promoter in all of the treated groups. Conclusion: Addiction with nicotine, morphine, methadone, and buprenorphine in doses and duration used in this study was not associated with the methylation of the Oprm1 promoter sites in the male Wistar rats.

Associations among the opioid receptor gene (OPRM1) A118G polymorphism, psychiatric symptoms, and quantitative EEG in Korean males with gambling disorder: A pilot study.

Background and aimsA single nucleotide polymorphism of A118G (SNP; rs1799971) in the opioid receptor μ-1 (OPRM1) gene is a missense variant that influences the affinity of μ-opioid receptors. This study aimed to investigate the associations among the A118G polymorphism in the OPRM1 gene, psychiatric symptoms, and quantitative electroencephalography (qEEG) findings in patients with gambling disorder.MethodsFifty-five male patients with gambling disorder aged between 18 and 65 years old participated in the study. The A118G polymorphism was genotyped into the AA, GA, and GG groups by the polymerase chain reaction/restriction fragment length polymorphism method. Resting-state qEEG was recorded with the eyes closed, and the absolute power of the delta (1–4 Hz), theta (4–8 Hz), alpha (8–12 Hz), and beta (12–30 Hz) frequency bands was analyzed. Psychiatric symptoms, including depression, anxiety, impulsivity and severity of gambling, were assessed by a self-rating scale.ResultsThere were no significant differences in psychiatric symptoms among the three genotype groups (AA, GA, and GG). However, the frequency band power of qEEG showed significant differences among the three genotype groups. The absolute power of the beta and theta bands in the frontal lobe was higher in G allele carriers.Discussion and conclusionBased on the findings of this study, the polymorphism in the OPRM1 gene might affect the neurophysiological process in patients with gambling disorder.

Increased risk of intracranial hemorrhage in preterm infants with OPRM1 gene A118G polymorphism.

To investigate the relationship between the OPRM1 gene A118G polymorphism and intracranial hemorrhage (ICH) in premature infants and identify the relevant genes in disease occurrence. In the present case study analysis, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the genotype and allele frequencies of the OPRM1 gene All8G single nucleotide polymorphism (SNP) in a case group of premature infants with ICH (n=167) and a control group of premature infants (n=163) without ICH. In the case group, 73 (43.7%) wild type A118 homozygous (A/A), 82 (49.1%) mutant heterozygous (A/G), and 12 (7.2%) mutant G118 homozygous (G/G) individuals were observed. The frequencies of A and G alleles were 68.3% and 31.7% respectively. In the control group, 89 (54.6%) wild type A118 homozygous (A/A), 68 (41.7%) mutant heterozygous (A/G), and 6 (3.7%) mutant G118 homozygous (G/G) individuals were observed. The frequencies of A and G alleles were 75.5% and 24.5% respectively. There was no significant difference in the frequency distribution of the OPRM1 gene A118G polymorphism between the two groups (χ2=4.839, P=0.089). There was a significant difference in the positive rate of wild-type AA and mutant-type (A/G + G/G) between the two groups (χ2=3.913, P=0.048). Carrying the G allele of the individual was 1.5 times more frequent suffering from the risk of ICH than carrying the A allele [odds ratio (OR): 1.549; 95% confidence interval (CI): 1.003-2.391], indicating that the OPRM1 118G allele was positively correlated with ICH and can increase the risk of ICH occurrence. The OPRM1 gene A118G polymorphism is associated with ICH in premature infants. The OPRM1 gene A118G may play a critical role in the occurrence of ICH.

Cross-talk between Human Spinal Cord μ-opioid Receptor 1Y Isoform and Gastrin-releasing Peptide Receptor Mediates Opioid-induced Scratching Behavior.

Although spinal opioids are safe and effective, pruritus is common and distressing. The authors previously demonstrated in mouse spinal cord that interactions between μ-opioid receptor isoform 1D and gastrin releasing peptide receptor mediate morphine-induced scratch. The C-terminal of 1D inhibits morphine-induced scratch without affecting analgesia. The authors hypothesize that human spinal cord also contains itch-specific μ-opioid receptor isoforms which interact with gastrin releasing peptide receptor. Reverse transcription polymerase chain reaction was performed on human spinal cord complimentary DNA from two human cadavers. Calcium responses to morphine (1 μM) were examined using calcium imaging microscopy on human cells (HEK293) coexpressing gastrin releasing peptide receptor and different human μ-opioid receptor isoforms. The authors assessed morphine-induced scratching behavior and thermal analgesia in mice following intrathecal injection of morphine (0.3 nmol) and a transactivator of transcription peptide designed from C-terminal sequences of 1Y isoform (0, 0.1, and 0.4 nmol). The authors demonstrated 1Y expression in the spinal cord dorsal horn. Morphine administration evoked a calcium response (mean ± SD) (57 ± 13 nM) in cells coexpressing both gastrin releasing peptide receptor and the 1Y isomer. This was blocked by 10 μM naltrexone (0.7 ± 0.4 nM; P < 0.0001), 1 μM gastrin-releasing peptide receptor antagonist (3 ± 2 nM; P < 0.0001), or 200 μM 1Y-peptide (2 + 2 nM; P < 0.0001). In mice, 0.4 nmol 1Y-peptide significantly attenuated morphine-induced scratching behaviors (scratching bouts, vehicle vs. 1Y-peptide) (92 ± 31 vs. 38 ± 29; P = 0.011; n = 6 to 7 mice per group), without affecting morphine antinociception in warm water tail immersion test (% of maximum possible effect) (70 ± 21 vs. 67 ± 22; P = 0.80; n = 6 mice per group). Human μ-opioid receptor 1Y isomer is a C-terminal splicing variant of Oprm1 gene identified in human spinal cord. Cross-talk between 1Y and gastrin releasing peptide receptor is required for mediating opioid-induced pruritus. Disrupting the cross talk may have implications for therapeutic uncoupling of desired analgesic effects from side effects of opioids.

Effects of mu-opioid receptor, ATP-banding cassette, subfamily B, member 1 gene, CYP3A genetic polymorphisms on sufentanil consumption in postoperative tumor patients

Objective To investigate the association between postoperative sufentanil consumption and genetic polymorphisms of mu-opioid receptor (OPRM1), ATP-banding cassette, subfamily B, member 1 gene(ABCB1) and CYP3A in postoperative tumor patients. Methods A total of one hundred and twenty patients [American Society of Anesthesiologists (ASA) Ⅰor Ⅱ, aging 19 to 65] who were scheduled to undergo tumor surgery under general anesthesia were enrolled in this study. Intravenous patient-controlled analgesia with sufentanil was provided postoperatively. Cumulative sufentanil consumption was measured 2, 6, 12, 24 h and 48 h after surgery. The severity of pain was assessed with the Visual Analogue Scale(VAS), while OPRM1 118A>G, ABCB1 2677G>A/T, ABCB1 3435C>T, and CYP3A4*1G and CYP3A5*3 variant alleles were detected. The effects of genetic and non-genetic factors on sufentanil requirements were evaluated with multiple linear regression analysis. Results Patient VAS scores 2, 6, 12, 24 h and 48 h after surgery were obviously associated with cumulative sufentanil doses (P<0.05). The 48 h cumulative sufentanil dose after surgery was associated with age(P<0.05). Conclusions There was no association between OPRM1, ABCB1, CYP3A genetic polymorphisms and postoperative sufentanil consumption in tumor patients. Key words: Pharmacogenetics; Sufentanil; CYP3A; Mu-opioid receptor; ATP-banding cassette, subfamily B, member 1 gene

Assessment of Effects of the OPRD1 and OPRM1 Genes Encoding Opioid Receptors on Apathy in Schizophrenia

Because of the involvement in motivational processes, opioid receptors are potential targets for apathy treatment in schizophrenia. We therefore searched for associations between the opioid receptor gene polymorphisms (OPRM1 (rs1799971) and OPRD1 (rs1042114, rs533123)) and apathy measured with the Apathy Evaluation Scale-S in a group of 284 schizophrenia patients. We analyzed individual genotypes, haplotypes, and the digenic interaction and observed nominally significant associations of rs1042114 genotypes and the rs1042114*rs1799971 interaction with behavioral apathy scores. The associations, however, did not withstand correction for multiple comparisons. Thus, the results did not provide enough evidence for the opioid receptor gene effects on apathy in schizophrenia patients.

Pharmacogenetics of Postoperative Nausea and Vomiting

Postoperative nausea and vomiting (PONV) remains one of the most common adverse effects of anesthesia, affecting up to 80% of high-risk patients within 24hours after surgery. Patient-related factors, surgical procedure, and perioperative medications such as opioids determine a patient's risk for PONV. To prevent and manage PONV, ondansetron, a 5-hydroxytryptamine type 3 (5-HT3) receptor antagonist, is frequently administered. Ondansetron is metabolized predominantly by hepatic cytochrome P450 (CYP2D6) enzymes, encoded by the CYP2D6 gene, whereas most of the effects of opioids are exerted at the opioid mu-1 receptor, encoded by the OPRM1 gene. Genetic polymorphisms of the CYP2D6 and OPRM1 genes may have a role in interindividual variation in the occurrence of PONV. Specifically, the occurrence of the G-allele produced by the OPRM1 A118G appears to be protective against PONV, whereas CYP2D6 ultrarapid metabolism increases the risk for PONV. The Clinical Pharmacogenetics Implementation Consortium guidelines provide CYP2D6-guided therapeutic recommendations for ondansetron. However, further studies are needed to investigate the role of genetic polymorphism in the occurrence of PONV and response to antiemetics.

Preclinical and Clinical Evidence for a Distinct Regulation of Mu Opioid and Type 1 Cannabinoid Receptor Genes Expression in Obesity.

Among endogenous signaling networks involved in both rewarding and homeostatic mechanisms of obesity, a relevant role is played by the endocannabinoid (ECS) and the opioid (EOS) systems. We here studied the transcriptional regulation of ECS and EOS genes in the hypothalamus of Diet-induced obesity rats, a preclinical model of obesity, as well as in humans with obesity and healthy controls. A significant and selective increase in type 1 cannabinoid receptor gene (Cnr1) expression was observed at the beginning of obesity development (5 weeks on high fat diet) as well as after 21 weeks of high diet exposure. After 5 weeks on high fat diet, selective up-regulation of mu opioid receptor gene (Oprm1) expression was also observed. Consistently, epigenetic studies showed a selective and significant decrease in DNA methylation at specific CpG sites at both gene promoters in overweight rats, but only after 5 weeks on high fat diet. Moreover, significantly lower levels of DNA methylation were observed at selected CpG sites of both receptor gene promoters, analyzed in peripheral blood mononuclear cells from younger (<30 years old) humans with obesity, as well as in those with shorter time length from disease onset. Taken together, we here provide evidence of selective, synergistic and time-dependent transcriptional regulation of CNR1 and OPRM1 genes in overweight rats, as well as in human subjects. These alterations in genes regulation could contribute to the development of the obese phenotype, and we thus suggest CNR1 and OPRM1 epigenetic modulation as possible biomarkers of obesity development. Due to the reversible nature of the epigenetic hallmark, our data might also open new avenue to early environmental strategies of intervention.

Synaptic Regulation by OPRM1 Variants in Reward Neurocircuitry.

Mu-opioid receptors (MORs) are the primary site of action of opioid drugs, both licit and illicit. Susceptibility to opioid addiction is associated with variants in the gene encoding the MOR, OPRM1 Varying with ethnicity, ∼25% of humans carry a single nucleotide polymorphism (SNP) in OPRM1 (A118G). This SNP produces a nonsynonymous amino acid substitution, replacing asparagine (N40) with aspartate (D40), and has been linked with an increased risk for drug addiction. While a murine model of human OPRM1 A118G (A112G in mouse) recapitulates most of the phenotypes reported in humans, the neuronal mechanisms underlying these phenotypes remain elusive. Here, we investigated the impact of A118G on opioid regulation of synaptic transmission in mesolimbic VTA dopaminergic neurons. Using electrophysiology, we showed that both inhibitory and excitatory inputs to VTA dopaminergic neurons projecting to the NAc medial shell were suppressed by the MOR agonists DAMGO and morphine, which caused a shift in the excitatory/inhibitory balance and an increased action potential firing rate. Mice carrying the 112G/G allele exhibited lower sensitivity to DAMGO and morphine compared with major allele carriers (112A/A). Paradoxically, DAMGO produced facilitatory effects on mEPSCs, which were mediated by presynaptic GABAB receptors. However, this was only prominent in homozygous major allele carriers, which could explain a stronger shift in action potential firing in 112A/A mice. This study provides a better understanding on the neurobiological mechanisms that may underlie risk of addiction development in carriers of the A118G SNP in OPRM1SIGNIFICANCE STATEMENT The pandemic of opioid drug abuse is associated with many socioeconomic burdens. The primary brain target of opioid drugs is the μ-opioid receptor (MOR), encoded by the OPRM1 gene, which is highly polymorphic in humans. Using a mouse model of the human OPRM1 A118G single nucleotide polymorphism (SNP) (A112G in mice), we demonstrated that MOR and GABAB signaling coordinate in regulating mesolimbic dopamine neuronal firing via presynaptic regulation. The A118G SNP affects MOR-mediated suppression of GABA and glutamate release, showing weaker efficacy of synaptic regulation by MORs. These results may shed light on whether MOR SNPs need to be considered for devising effective therapeutic interventions.

The prevalence of ADH1B and OPRM1 alleles predisposing for alcohol consumption are increased in the Hungarian psoriasis population

Alcohol intake affects in great the symptoms and life of psoriasis patients, although the association of SNPs related to increased alcohol consumption with psoriasis has not been elucidated. Therefore, to investigate the association of psoriasis with established alcohol consumption and dependence-related gene variants we conducted a population-based case–control study including 3743 subjects (776 psoriasis cases and 2967 controls from the general Hungarian population). Genotyping of 23 SNPs at ADH1B, ADH1C, ALDH1A1, ALDH2, SLC6A3, DDC, GABRA2, GABRG1, HTR1B, MAOA, TPH2, CHRM2, GRIN2A, POMC, OPRM1, OPRK1 and BDNF were determined and differences in genotype and allele distributions were investigated. Multiple logistic regression analyses were implemented. Analysis revealed association between C allele of the rs1229984 polymorphism (ADH1B gene) and psoriasis risk (ORadditive = 1.58, 95% CI 1.23–2.03, p < 0.001, ORrecessive = 1.58, 95% CI 1.22–2.04, p = 0.001). Furthermore, the G allele of rs1799971 polymorphism (OPRM1 gene) increased the risk of familial aggregation (ORadditive = 1.99, 95% CI 1.36–2.91, p < 0.001 ORdominant = 2.01, 95% CI 1.35–3.01, p < 0.001). In subgroups of psoriatic patients with history of early onset and familial aggregation effect allele ‘C’ of rs1229984 showed association in the additive and recessive models (ORadditive = 2.41, 95% CI 1.26–4.61, p < 0.01, ORrecessive = 2.42, 95% CI 1.26–4.68, p < 0.01). While effect allele ‘G’ of rs1799971 (OPRM1) also associated with increased risk of early onset and familial aggregation of psoriasis in the additive and dominant models (ORadditive = 1.75, 95% CI 1.27–2.43, p = 0.001, ORdominant = 1.82, 95% CI 1.26–2.63, p = 0.001). Our results suggest that genetically defined high-risk individuals for alcohol consumption are more common in the psoriasis population.

Mu opioid receptors in the medial habenula contribute to naloxone aversion.

The medial habenula (MHb) is considered a brain center regulating aversive states. The mu opioid receptor (MOR) has been traditionally studied at the level of nociceptive and mesolimbic circuits, for key roles in pain relief and reward processing. MOR is also densely expressed in MHb, however, MOR function at this brain site is virtually unknown. Here we tested the hypothesis that MOR in the MHb (MHb-MOR) also regulates aversion processing. We used chnrb4-Cre driver mice to delete the Oprm1 gene in chnrb4-neurons, predominantly expressed in the MHb. Conditional mutant (B4MOR) mice showed habenula-specific reduction of MOR expression, restricted to chnrb4-neurons (50% MHb-MORs). We tested B4MOR mice in behavioral assays to evaluate effects of MOR activation by morphine, and MOR blockade by naloxone. Locomotor, analgesic, rewarding, and motivational effects of morphine were preserved in conditional mutants. In contrast, conditioned place aversion (CPA) elicited by naloxone was reduced in both naïve (high dose) and morphine-dependent (low dose) B4MOR mice. Further, physical signs of withdrawal precipitated by either MOR (naloxone) or nicotinic receptor (mecamylamine) blockade were attenuated. These data suggest that MORs expressed in MHb B4-neurons contribute to aversive effects of naloxone, including negative effect and aversive effects of opioid withdrawal. MORs are inhibitory receptors, therefore we propose that endogenous MOR signaling normally inhibits chnrb4-neurons of the MHb and moderates their known aversive activity, which is unmasked upon receptor blockade. Thus, in addition to facilitating reward at several brain sites, tonic MOR activity may also limit aversion within the MHb circuitry.

The role of OPRM1 polymorphism in the etiology of alcoholism

Numerous studies have investigated the association between the OPRM1 A118G polymorphism (rs1799971) and alcohol dependence, but the results have been inconsistent. The endogenous opioid system has been implicated in the development of alcohol dependence for its prominent role in the central rewarding mechanism. The aim of this study was to evaluate the role of the A118G polymorphism of the OPRM1 gene in the pathogenesis of alcohol dependence syndrome (ADS). The OPRM1 (rs1799971) polymorphism was investigated in an association study of a group of ADS patients (n = 177) and in subgroups (delirium tremens and/or seizures, age at onset <26 years, dissocial alcoholics, positive familial history of alcoholism, delirium tremens, and seizures). The control group consisted of healthy volunteers, with matched gender and age, and with psychiatric disorders excluded (n = 162). Our research shows that there are differences in the genotypes and alleles of the OPRM1 polymorphism in the case-control study. Furthermore, we observed associations in our homogeneous subgroups - in the group of patients with ADS and accompanying delirium tremens and/or seizures at the genotype level, as well as in the subgroup of patients under 26 years of age with an early onset of dependence. It is strongly possible that the G allele described in numerous studies can be associated with a response to treatment, but not typology, or the very predisposition toward alcoholism. It is necessary to carry out further research which would embrace a larger group of patients; it should be divided into other homogeneous subgroups, including, e.g., naltrexone pharmacotherapy.

Binge-Like Eating Is Not Influenced by the Murine Model of OPRM1 A118G Polymorphism.

Impairments in opioid receptor signaling have been implicated in disordered eating. A functional variant of the OPRM1 gene is a guanine (G) substitution for adenine (A) at the 118 position of exon 1 (A118G). The influence of the A118G variant on binge eating behaviors and the effectiveness of pharmacotherapies used to treat binge eating have not been characterized. Mice were generated with A to G substitution at the 112 position on exon 1 to produce a murine equivalent of the human A118G variant. Homozygous female mice (AA or GG) were exposed to intermittent access to a highly palatable sweet-fat food with or without prior calorie deprivation to promote dietary-induced binge eating. There were no genotype-dependent differences in the dietary-induced binge eating. However, GG mice exposed to intermittent calorie restriction (Restrict) had higher body weights compared with GG mice exposed to intermittent sweet fat-food (Binge) and ad libitum feeding (Naive). Acute oral dosing of lisdexamfetamine (0.15, 0.5, and 1.5 mg/kg) or sibutramine (0.3, 1, and 3 mg/kg) did not produce genotype-dependent differences in binge-like eating. In addition, no genotype-dependent differences in binge-like eating were observed with chronic (14-day) dosing of lisdexamfetamine (1.5 mg/kg/day) or sibutramine (3 mg/kg/day). In the chronic dosing, body weights were higher in the GG Restrict compared with AA Restrict. Our findings suggest that the A112G polymorphism does not influence binge eating behaviors or pharmacotherapies for treating binge eating.

μ-Opioid receptors in primary sensory neurons are essential for opioid analgesic effect on acute and inflammatory pain and opioid-induced hyperalgesia.

μ-Opioid receptors (MORs) are expressed peripherally and centrally, but the loci of MORs responsible for clinically relevant opioid analgesia are uncertain. Crossing Oprm1flox/flox and AdvillinCre/+ mice completely ablates MORs in dorsal root ganglion neurons and reduces the MOR expression level in the spinal cord. Presynaptic MORs expressed at primary afferent central terminals are essential for synaptic inhibition and potentiation of sensory input by opioids. MOR ablation in primary sensory neurons diminishes analgesic effects produced by systemic and intrathecal opioid agonists and abolishes chronic opioid treatment-induced hyperalgesia. These findings demonstrate a critical role of MORs expressed in primary sensory neurons in opioid analgesia and suggest new strategies to increase the efficacy and reduce adverse effects of opioids. The pain and analgesic systems are complex, and the actions of systemically administered opioids may be mediated by simultaneous activation of μ-opioid receptors (MORs, encoded by the Oprm1 gene) at multiple, interacting sites. The loci of MORs and circuits responsible for systemic opioid-induced analgesia and hyperalgesia remain unclear. Previous studies using mice in which MORs are removed from Nav1.8- or TRPV1-expressing neurons provided only an incomplete and erroneous view about the role of peripheral MORs in opioid actions in vivo. In the present study, we determined the specific role of MORs expressed in primary sensory neurons in the analgesic and hyperalgesic effects produced by systemic opioid administration. We generated Oprm1 conditional knockout (Oprm1-cKO) mice in which MOR expression is completely deleted from dorsal root ganglion neurons and substantially reduced in the spinal cord, which was confirmed by immunoblotting and immunocytochemical labelling. Both opioid-induced inhibition and potentiation of primary sensory input were abrogated in Oprm1-cKO mice. Remarkably, systemically administered morphine potently inhibited acute thermal and mechanical nociception and persistent inflammatory pain in control mice but had little effect in Oprm1-cKO mice. The analgesic effect of intrathecally administered morphine was also profoundly reduced in Oprm1-cKO mice. Additionally, chronic morphine treatment-induced hyperalgesia was absent in Oprm1-cKO mice. Our findings directly challenge the notion that clinically relevant opioid analgesia is mediated mostly by centrally expressed MORs. MORs in primary sensory neurons, particularly those expressed presynaptically at the first sensory synapse in the spinal cord, are crucial for both opioid analgesia and opioid-induced hyperalgesia.

OPRM1 Gene Interaction with Sleep in Chronic Pain Patients Treated with Opioids

Background: The experience of chronic non-cancer pain (CNCP) is one of the most common reasons individuals seek medical attention. Patients with CNCP frequently experience concomitant sleep-related problems. Objectives: The aim was to evaluate sleep problems in opioid naïve CNCP patients, before and after opioid titration, analyzing the influence of OPRM1 gene variants. Study Design: A prospective, cohort, observational study. Setting: This study was performed at the Pain Unit of the Alicante University General Hospital. Methods: Pain and Medical Outcomes Study Sleep questionnaire (MOS-Sleep) were assessed at baseline and 3 months after opioid titration in 231 opioid naïve CNCP patients. Sleep data was compared with a matched-control group (n = 64). Morphine equivalent daily doses, adverse events, and drugs prescribed for pain were also registered. OPRM1 polymorphism rs1799971 was analyzed by RT-PCR. Ethics Committee approved the study and results were analyzed by R software. Results: After 3 months of opioid titration, patients with CNCP (63 ± 14 years, 64% female, VAS 74 ± 17 mm) significantly decreased pain intensity, anxiety and depression, and increased quality of life. Sleep problems were significantly more frequent in females (P = 0.002). Age, quality of life, anxiety, and depression all influenced sleep disturbances and problems indices, which were significantly different from the control group. Furthermore, the OPRM1 118- GG genotype was also associated with significantly lower sleep adequacy, and more sleep problems. Limitations: Total number of subjects studied was relatively small and most patients were on other non-opioid centrally-acting medications. Conclusions: Opioids decreased CNCP severity, improving patients’ psychological areas, and quality of life. However, patients with OPRM1 118-GG genotype indicated an increase in sleep problems and worsening sleep pattern while taking opioids. Key words: OPRM1, pharmacogenetics, MOS-Sleep, opioids, chronic noncancer pain, sleep related problems, sleep problem index SLP-6 and SLP-9