Rate Of Nicotine Metabolism Research Articles

Racial differences in the relationship between rate of nicotine metabolism and nicotine intake from cigarette smoking

Rate of nicotine metabolism has been identified as an important factor influencing nicotine intake and can be estimated using the nicotine metabolite ratio (NMR), a validated biomarker of CYP2A6 enzyme activity. Individuals who metabolize nicotine faster (higher NMR) may alter their smoking behavior to titrate their nicotine intake in order to maintain similar levels of nicotine in the body compared to slower nicotine metabolizers. There are known racial differences in the rate of nicotine metabolism with African Americans on average having a slower rate of nicotine metabolism compared to Whites. The goal of this study was to determine if there are racial differences in the relationship between rate of nicotine metabolism and measures of nicotine intake assessed using multiple biomarkers of nicotine and tobacco smoke exposure. Using secondary analyses of the screening data collected in a recently completed clinical trial, treatment-seeking African American and White daily smokers (10 or more cigarettes per day) were grouped into NMR quartiles so that the races could be compared at the same NMR, even though the distribution of NMR within race differed. The results indicated that rate of nicotine metabolism was a more important factor influencing nicotine intake in White smokers. Specifically, Whites were more likely to titrate their nicotine intake based on the rate at which they metabolize nicotine. However, this relationship was not found in African Americans. Overall there was a greater step-down, linear type relationship between NMR groups and cotinine or cotinine/cigarette in African Americans, which is consistent with the idea that differences in blood cotinine levels between the African American NMR groups were primarily due to differences in CYP2A6 enzyme activity without titration of nicotine intake among faster nicotine metabolizers.

Cessation of alcohol consumption decreases rate of nicotine metabolism in male alcohol-dependent smokers

BackgroundRate of nicotine metabolism is an important factor influencing cigarette smoking behavior, dependence, and efficacy of nicotine replacement therapy. The current study examined the hypothesis that chronic alcohol abuse can accelerate the rate of nicotine metabolism. Nicotine metabolite ratio (NMR, a biomarker for rate of nicotine metabolism) and patterns of nicotine metabolites were assessed at three time points after alcohol cessation. MethodsParticipants were 22 Caucasian men randomly selected from a sample of 165 smokers entering a 7-week alcohol dependence treatment program in Poland. Data were collected at three time points: baseline (week 1, after acute alcohol detoxification), week 4, and week 7. Urine was analyzed for nicotine and metabolites and used to determine the nicotine metabolite ratio (NMR, a biomarker for rate of nicotine metabolism), and total nicotine equivalents (TNE, a biomarker for total daily nicotine exposure). Results and conclusionsThere was a significant decrease in urine NMR over the 7 weeks after alcohol abstinence (F(2,42)=18.83, p<0.001), indicating a decrease in rate of nicotine metabolism. On average NMR decreased 50.0% from baseline to week 7 (9.6±1.3 vs 4.1±0.6). There was no change in urine TNE across the three sessions, indicating no change daily nicotine intake. The results support the idea that chronic alcohol abuse may increase the rate of nicotine metabolism, which then decreases over time after alcohol cessation. This information may help to inform future smoking cessation interventions in this population.

Brain Responses to Smoking Cues Differ Based on Nicotine Metabolism Rate

BackgroundInherited differences in the rate of metabolism of nicotine, the addictive chemical in tobacco, affect smoking behavior and quitting success. The nicotine metabolite ratio (3′-hydroxycotinine/cotinine) is a reliable measure of nicotine clearance and a well-validated predictive biomarker of response to pharmacotherapy. To clarify the mechanisms underlying these associations, we investigated the neural responses to smoking cues in normal and slow nicotine metabolizers. MethodsTreatment-seeking smokers (N = 69; 30 slow metabolizers and 39 normal metabolizers) completed a visual cue reactivity task during functional magnetic resonance imaging on two separate occasions: once during smoking satiety and once after 24 hours of smoking abstinence. ResultsIn whole-brain analysis, normal (compared with slow) metabolizers exhibited heightened abstinence-induced neural responses to smoking cues in the left caudate, left inferior frontal gyrus, and left frontal pole. These effects were more pronounced when extreme groups of slow and normal metabolizers were examined. Greater activation in the left caudate and left frontal pole was associated with abstinence-induced subjective cravings to smoke. ConclusionsInherited differences in rate of nicotine elimination may drive neural responses to smoking cues during early abstinence, providing a plausible mechanism to explain differences in smoking behaviors and response to cessation treatment. Normal metabolizers may benefit from adjunctive behavioral smoking cessation treatments, such as cue exposure therapy.

CYP2A6 Effects on Subjective Reactions to Initial Smoking Attempt.

In very novice smokers, CYP2A6 genotypes that reduce nicotine metabolism to an intermediate rate may increase smoking risk, relative to both normal and slow rates. The present study examined the hypothesis that intermediate metabolism variants are associated with greater pleasurable effects of the initial smoking attempt than either normal or slow metabolism variants. Participants were novice smokers (N = 261, 65% female) of European descent. Predicted nicotine metabolic rate based on CYP2A6 diplotypes (CYP2A6 Diplotype Predicted Rate [CDPR]) was partitioned into Normal, Intermediate, and Slow categories using a metabolism metric. Subjective reactions to the initial smoking attempt were assessed by the Pleasurable Smoking Experiences (PSE) scale, which was collected within 3 years of the initial smoking attempt. The effect of CDPR on PSE was tested using a generalized linear model in which CDPR was dummy coded and Intermediate CDPR was the reference condition. Gender was included in the model as a control for higher PSE scores by males. Lower PSE scores were associated with Normal CDPR, β = -0.34, P = .008, and Slow CDPR, β = -0.52, P = .001, relative to Intermediate CDPR. Intermediate CDPR-enhanced pleasurable effects of the initial smoking attempt relative to other CYP2A6 variants. This finding is consistent with the hypothesis that the risk effect of Intermediate CDPR on early smoking is a function of optimal pleasurable effects. This study supports our recent hypothesis that CYP2A6 diplotypes that encode intermediate nicotine metabolism rate are associated with enhanced pleasurable events following the initial smoking attempt, compared with diplotypes that encode either normal or slow metabolism. This hypothesis was offered to account for our unexpected previous finding of enhanced smoking risk in very novice smokers associated with intermediate metabolism rate. Our new finding encourages further investigation of time-dependent relations between CYP2A6 effects and smoking motives, and it encourages laboratory study of the mechanisms underlying the initial smoking enhancement in novice smokers associated with intermediate metabolism.

Abstract IA47: Resolving the complex problem of tobacco-caused lung cancer disparities in the U.S

Abstract Existing data show that lung cancer disparities are not explained by cigarette smoking dose and duration alone. Data from the Multiethnic Cohort Study show that Native Hawaiians and African Americans who smoked 10 cigarettes per day have an elevated risk of lung cancer compared to Japanese American, White, and Latino smokers. Prior studies also suggest that African Americans have slower nicotine metabolism compared to Whites. It remains unclear why we consistently observe slower rates of nicotine metabolism among African Americans, but higher rates of lung cancer since studies suggest that smokers with low levels of CPY2A6 activity may be less efficient in bioactivating tobacco smoke pre-carcinogens to carcinogens. Little is known about nicotine metabolism among Native Hawaiians and Filipinos who have disproportionate and unexplained lung cancer rates like African Americans. This study compares biomarkers of tobacco smoke exposure in Native Hawaiian, Filipino, and White young adult daily smokers. We hypothesized that Native Hawaiians and Filipinos, like African Americans, would have slower nicotine metabolism compared to Whites. We collected data on sociodemographics, smoking history, and psychosocial risk factors among young adult daily smokers aged 18-35. We measured height, weight, and carbon monoxide levels among all smokers. A saliva sample was collected from each smoker using standard passive drool procedures. The geometric means were calculated for nicotine, cotinine, trans 3' hydroxycotinine, the nicotine metabolite ratio, and expired carbon monoxide and the data were compared among racial/ethnic groups. Two analysis of covariance models tested biomarker differences among racial/ethnic groups. Model 1, the unadjusted model, contained no covariates. Model 2 included gender, body mass index (BMI), menthol smoking status, Hispanic ethnicity, and the number of cigarettes smoked per day as covariates. The sample included 44% Native Hawaiians, 16% Filipinos, and 40% Whites (n=186). Twenty-four percent of young adults were of Hispanic origin, 48% were females, and smokers had a mean BMI of 28. Forty-one percent of Filipino smokers reported Hispanic ethnicity compared to 27% of Native Hawaiian and 15% of White smokers (p&amp;lt; 0.02). Native Hawaiians had a higher BMI than Filipinos and Whites (31.9, 24.5, 24.8, p&amp;lt;0.001). Eighty-seven percent of Native Hawaiians, 72% of Filipinos, and 48% of White young adults reported menthol cigarette use. There were no differences in smoking duration or smoking consumption among racial/ethnic groups. Smokers had smoked daily for 1.5 years and smoked a mean of 14.4 cigarettes per day. In the unadjusted and adjusted models, cotinine levels were higher among Native Hawaiians compared to Whites, but these differences were not significant among racial/ethnic groups. In the unadjusted model, the nicotine metabolic ratio was significantly lower in Native Hawaiian and Filipino smokers compared to White smokers (NMR: 0.17, 0.15, 0.27, p&amp;lt;0.001). In the adjusted model, the nicotine metabolite ratio remained significantly lower in Native Hawaiian and Filipino smokers compared to White smokers (NMR: 0.20, 0.19, 0.33, p&amp;lt;0.001). In summary, our data show that Native Hawaiian and Filipino daily smokers have slower nicotine metabolism than White daily smokers as indicated by the nicotine metabolite ratio. Studies show that the nicotine metabolite ratio is a reliable phenotypic marker for CYP2A6 activity. However, is it not clear whether the algorithm suggesting that slow nicotine metabolizers have reduced lung cancer risk applies to all racial/ethnic groups. Resolving the complex problem of lung cancer disparities requires further investigation of multiple biological pathways to determine which lung cancer risk algorithms apply to specific racial/ethnic groups. Such investigations will allow to us to improve community, clinical, and policy-based interventions to reduce disparities. Citation Format: Pebbles Fagan, Eric T. Moolchan, Pallav Pokhrel, Thaddeus Herzog, Kevin Cassel, Ian Pagano, Adrian Franke, Joseph Keawe'aimoku Kaholokula, Angela Sy, Linda A. Alexander, Dennis R. Trinidad, Kari-Lyn Sakuma, Carl Anderson Johnson, Alyssa M. Antonio, Dorothy Jorgensen, Tania Lynch, Crissy Kawamoto, Mark S. Clanton. Resolving the complex problem of tobacco-caused lung cancer disparities in the U.S. [abstract]. In: Proceedings of the Seventh AACR Conference on The Science of Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; Nov 9-12, 2014; San Antonio, TX. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2015;24(10 Suppl):Abstract nr IA47.

A Genome-Wide Association Study of a Biomarker of Nicotine Metabolism.

Individuals with fast nicotine metabolism typically smoke more and thus have a greater risk for smoking-induced diseases. Further, the efficacy of smoking cessation pharmacotherapy is dependent on the rate of nicotine metabolism. Our objective was to use nicotine metabolite ratio (NMR), an established biomarker of nicotine metabolism rate, in a genome-wide association study (GWAS) to identify novel genetic variants influencing nicotine metabolism. A heritability estimate of 0.81 (95% CI 0.70–0.88) was obtained for NMR using monozygotic and dizygotic twins of the FinnTwin cohort. We performed a GWAS in cotinine-verified current smokers of three Finnish cohorts (FinnTwin, Young Finns Study, FINRISK2007), followed by a meta-analysis of 1518 subjects, and annotated the genome-wide significant SNPs with methylation quantitative loci (meQTL) analyses. We detected association on 19q13 with 719 SNPs exceeding genome-wide significance within a 4.2 Mb region. The strongest evidence for association emerged for CYP2A6 (min p = 5.77E-86, in intron 4), the main metabolic enzyme for nicotine. Other interesting genes with genome-wide significant signals included CYP2B6, CYP2A7, EGLN2, and NUMBL. Conditional analyses revealed three independent signals on 19q13, all located within or in the immediate vicinity of CYP2A6. A genetic risk score constructed using the independent signals showed association with smoking quantity (p = 0.0019) in two independent Finnish samples. Our meQTL results showed that methylation values of 16 CpG sites within the region are affected by genotypes of the genome-wide significant SNPs, and according to causal inference test, for some of the SNPs the effect on NMR is mediated through methylation. To our knowledge, this is the first GWAS on NMR. Our results enclose three independent novel signals on 19q13.2. The detected CYP2A6 variants explain a strikingly large fraction of variance (up to 31%) in NMR in these study samples. Further, we provide evidence for plausible epigenetic mechanisms influencing NMR.

Decreased Nicotinic Receptor Availability in Smokers with Slow Rates of Nicotine Metabolism.

The nicotine metabolite ratio (NMR), a stable measure of hepatic nicotine metabolism via the CYP2A6 pathway and total nicotine clearance, is a predictive biomarker of response to nicotine replacement therapy, with increased quit rates in slower metabolizers. Nicotine binds directly to nicotinic acetylcholine receptors (nAChRs) to exert its psychoactive effects. This study examined the relationship between NMR and nAChR (α4β2* subtype) availability using PET imaging of the radiotracer 2-(18)F-fluoro-3-(2(S)-azetidinylmethoxy)pyridine (2-(18)F-FA-85380, or 2-(18)F-FA). Twenty-four smokers-12 slow metabolizers (NMR < 0.26) and 12 normal metabolizers (NMR ≥ 0.26)-underwent 2-(18)F-FA-PET brain imaging after overnight nicotine abstinence (18 h before scanning), using a validated bolus-plus-infusion protocol. Availability of nAChRs was compared between NMR groups in a priori volumes of interest, with total distribution volume (VT/fP) being the measure of nAChR availability. Cravings to smoke were assessed before and after the scans. Thalamic nAChR α4β2* availability was significantly reduced in slow nicotine metabolizers (P = 0.04). Slow metabolizers exhibited greater reductions in cravings after scanning than normal metabolizers; however, craving was unrelated to nAChR availability. The rate of nicotine metabolism is associated with thalamic nAChR availability. Additional studies could examine whether altered nAChR availability underlies the differences in treatment response between slow and normal metabolizers of nicotine.

Changes in the rate of nicotine metabolism across pregnancy: a longitudinal study.

AimsIncreased nicotine metabolism during pregnancy could explain why nicotine replacement therapy (NRT) appears to be less effective on smoking cessation in pregnancy than in non‐pregnant smokers, but little is known about nicotine metabolism across pregnancy. This study was conducted to determine when changes in nicotine metabolism occur during pregnancy and to describe the magnitude of these changes.DesignLongitudinal cohort study of pregnant smokers' nicotine metabolite ratio (NMR).Setting and Participants101 pregnant smokers recruited from hospital antenatal clinics in Nottingham, UK were asked to provide saliva samples at 8–14 weeks (n = 98), 18–22 weeks (n = 65), 32–36 weeks gestation (n = 47), 4 weeks postpartum (n = 44) and 12 weeks postpartum (n = 47).MeasurementsNicotine metabolite ratio (NMR) was measured using the ratio of cotinine to its primary metabolite trans‐3'‐hydroxycotinine. Multi‐level modelling was used to detect any overall difference in NMR between time points. The 12 week postpartum NMR was compared with the NMRs collected antenatally and 4 weeks postpartum.FindingsNMR changed over time (p = 0.0006). Compared with NMR at 12 weeks postpartum, NMR was significantly higher at 18–22 weeks (26% higher, 95% CI 12% to 38%) and 32–36 weeks (23% higher, 95% CI 9% to 35%). There was no significant difference between the 8‐14 weeks gestation or 4 weeks postpartum NMR and 12 weeks postpartum.ConclusionsNicotine metabolism appears to be faster during pregnancy; this faster metabolism is apparent from 18 to 22 weeks of pregnancy and appears to fall by 4 weeks after childbirth.

Nicotine metabolite ratio (3-hydroxycotinine/cotinine) in plasma and urine by different analytical methods and laboratories: implications for clinical implementation.

The highly genetically variable enzyme CYP2A6 metabolizes nicotine to cotinine (COT) and COT to trans-3'-hydroxycotinine (3HC). The nicotine metabolite ratio (NMR, 3HC/COT) is commonly used as a biomarker of CYP2A6 enzymatic activity, rate of nicotine metabolism, and total nicotine clearance; NMR is associated with numerous smoking phenotypes, including smoking cessation. Our objective was to investigate the impact of different measurement methods, at different sites, on plasma and urinary NMR measures from ad libitum smokers. Plasma (n = 35) and urine (n = 35) samples were sent to eight different laboratories, which used similar and different methods of COT and 3HC measurements to derive the NMR. We used Bland-Altman analysis to assess agreement, and Pearson correlations to evaluate associations, between NMR measured by different methods. Measures of plasma NMR were in strong agreement between methods according to Bland-Altman analysis (ratios, 0.82-1.16) and were highly correlated (all Pearson r > 0.96, P < 0.0001). Measures of urinary NMR were in relatively weaker agreement (ratios 0.62-1.71) and less strongly correlated (Pearson r values of 0.66-0.98, P < 0.0001) between different methods. Plasma and urinary COT and 3HC concentrations, while weaker than NMR, also showed good agreement in plasma, which was better than that in urine, as was observed for NMR. Plasma is a very reliable biologic source for the determination of NMR, robust to differences in these analytical protocols or assessment site. Together this indicates a reduced need for differential interpretation of plasma NMR results based on the approach used, allowing for direct comparison of different studies.

Rate of Nicotine Metabolism and Smoking Cessation Outcomes in a Community-based Sample of Treatment-Seeking Smokers

BackgroundIn samples from controlled randomized clinical trials, a smoker’s rate of nicotine metabolism, measured by the 3-hydroxycotinine to cotinine ratio (NMR), predicts response to transdermal nicotine. Replication of this relationship in community-based samples of treatment-seeking smokers may help guide the implementation of the NMR for personalized treatment for nicotine dependence. MethodsData from a community-based sample of treatment seeking smokers (N=499) who received 8weeks of transdermal nicotine and 4 behavioral counseling sessions were used to evaluate associations between the NMR and smoking cessation. Secondary outcomes included withdrawal and craving, depression and anxiety, side effects, and treatment adherence. ResultsThe NMR was a significant predictor of abstinence (OR=.56, 95% CI: 0.33-0.95, p=.03), with faster metabolizers showing lower quit rates than slower metabolizers (24% vs. 33%). Faster nicotine metabolizers exhibited significantly higher levels of anxiety symptoms over time during treatment, vs. slower metabolizers (NMR x Time interaction: F[3,357]=3.29, p=.02). NMR was not associated with changes in withdrawal, craving, depression, side effects, and treatment adherence (p’s>.05). ConclusionsIn a community-based sample of treatment-seeking smokers, faster nicotine metabolizers were significantly less likely to quit smoking and showed higher rates of anxiety symptoms during a smoking cessation treatment program, vs. slower nicotine metabolizers. These results provide further evidence that transdermal nicotine is less effective for faster nicotine metabolizers and suggest the need to address cessation-induced anxiety symptoms among these smokers to increase the chances for successful smoking cessation.

Lack of Associations of CHRNA5-A3-B4 Genetic Variants with Smoking Cessation Treatment Outcomes in Caucasian Smokers despite Associations with Baseline Smoking.

CHRNA5-A3-B4 variants, rs16969968, rs588765 and rs578776, are consistently associated with tobacco consumption among smokers, but the association with smoking cessation is less consistent. Among the studies that reported significant associations with cessation, the effects were observed in smokers treated with placebo treatment in some studies and conversely in those receiving active pharmacological therapy (bupropion and nicotine replacement therapies) in others. Thus, it remains unclear whether CHRNA5-A3-B4 is a useful marker for optimizing smoking cessation. Using data from 654 Caucasian smokers treated with placebo, nicotine patch or varenicline, we investigated whether CHRNA5-A3-B4 variants were associated with smoking cessation outcomes, and whether there were significant genotype-by-treatment or haplotype-by-treatment interactions. We observed no significant associations between CHRNA5-A3-B4 variants and smoking cessation, despite replicating previous associations with baseline tobacco consumption. At end of treatment the effect size on smoking cessation in the placebo, patch and varenicline groups for rs16969968 [GG vs. GA+AA] was OR = 0.66 (P = 0.23), OR = 1.01 (P = 0.99), and OR = 1.30 (P = 0.36) respectively, of rs588765 [CC vs. CT+TT] was OR = 0.96 (P = 0.90), OR = 0.84 (P = 0.58), and OR = 0.74 (P = 0.29) respectively, and for rs578776 [GG vs. GA+AA] on smoking cessation was OR = 1.02 (P = 0.95), OR = 0.75 (P = 0.35), and OR = 1.20 (P = 0.51) respectively. Furthermore, we observed no associations with cessation using the CHRNA5-A3-B4 haplotype (constructed using rs16969968 and rs588765), nor did we observe any significant genotype-by-treatment interactions, with or without adjusting for the rate of nicotine metabolism (all P>0.05). We also observed no significant genetic associations with 6 month or 12 month smoking abstinence. In conclusion, we found no association between CHRNA5-A3-B4 variants and smoking cessation rates in this clinical trial; however, as expected, significant associations with baseline tobacco consumption were replicated. Our data suggest that CHRNA5-A3-B4 gene variants do not exhibit a robust association with smoking cessation and are unlikely to be useful for clinically optimizing smoking cessation pharmacotherapy for Caucasian smokers.

Nicotine Metabolism in Young Adult Daily Menthol and Nonmenthol Smokers.

Menthol cigarette smoking may increase the risk for tobacco smoke exposure and inhibit nicotine metabolism in the liver. Nicotine metabolism is primarily mediated by the enzyme CYP2A6 and the nicotine metabolite ratio (NMR = trans 3' hydroxycotinine/cotinine) is a phenotypic proxy for CYP2A6 activity. No studies have examined differences in this biomarker among young adult daily menthol and nonmenthol smokers. This study compares biomarkers of tobacco smoke exposure among young adult daily menthol and nonmenthol smokers. Saliva cotinine and carbon monoxide were measured in a multiethnic sample of daily smokers aged 18-35 (n = 186). Nicotine, cotinine, the cotinine/cigarette per day ratio, trans 3' hydroxycotinine, the NMR, and expired carbon monoxide were compared. The geometric means for nicotine, cotinine, and the cotinine/cigarette per day ratio did not significantly differ between menthol and nonmenthol smokers. The NMR was significantly lower among menthol compared with nonmenthol smokers after adjusting for race/ethnicity, gender, body mass index, and cigarette smoked per day (0.19 vs. 0.24, P = .03). White menthol smokers had significantly higher cotinine/cigarettes per day ratio than white nonmenthol smokers in the adjusted model. White menthol smokers had a lower NMR in the unadjusted model (0.24 vs. 0.31, P = .05) and the differences remained marginally significant in the adjusted model (0.28 vs. 0.34, P = .06). We did not observe these differences in Native Hawaiians and Filipinos. Young adult daily menthol smokers have slower rates of nicotine metabolism than nonmenthol smokers. Studies are needed to determine the utility of this biomarker for smoking cessation treatment assignments.

Nicotine Metabolite Ratio Is Associated With Lozenge Use But Not Quitting in Smokeless Tobacco Users.

The nicotine metabolite ratio (NMR) of 3'-hydroxycotinine to cotinine is a noninvasive marker of the rate of nicotine metabolism. Fast metabolism (ie, a high NMR) is associated with lower cigarette smoking abstinence rates using transdermal nicotine replacement. We evaluated whether the NMR can be used to predict self-reported nicotine lozenge use and tobacco abstinence among smokeless tobacco users treated for tobacco dependence. This was a secondary analysis of data from one arm of a large trial. Participants received quitting support materials and 4-mg nicotine lozenges by mail plus three coaching phone calls. Saliva kits were mailed for collection of saliva samples, which were analyzed for cotinine and 3'-hydroxycotinine. Self-reported tobacco and lozenge use were assessed at 3 months. Analyses were performed using Spearman rank correlation and logistic regression. Of the 160 saliva collection kits mailed, 152 were returned. The NMR was not significantly correlated with the baseline amount of smokeless tobacco used, the number of years of tobacco use, or the level of tobacco dependence as measured by the Severson Smokeless Tobacco Dependency Scale. The NMR was positively correlated with lozenge use (r = 0.21, P = .015), but it did not predict self-reported 7-day point prevalence abstinence at 3 months. Fast metabolizers may need to self-administer more nicotine replacement in the form of nicotine lozenges to achieve the same clinical response achieved by slower metabolizers using fewer lozenges.

CYP2A6 Longitudinal Effects in Young Smokers.

The present study sought to identify time-dependent within-participant effects of CYP2A6 genotypes on smoking frequency and nicotine dependence in young smokers. Predicted nicotine metabolic rate based on CYP2A6 diplotypes (CYP2A6 diplotype predicted rate [CDPR]) was partitioned into Normal, Intermediate, and Slow categories using a metabolism metric. Growth-curve models characterized baseline and longitudinal CDPR effects with data from eight longitudinal assessments during a 6-year period (from approximately age 16-22) in young smokers of European descent (N = 296, 57% female) who had smoked less than 100 cigarettes lifetime at baseline and more than that amount by Year 6. Phenotypes were number of days smoked during the previous 30 days and a youth version of the Nicotine Dependence Syndrome Scale (NDSS). A zero-inflated Poisson growth-curve model was used to account for the preponderance of zero days smoked. At baseline, Intermediate CDPR was a risk factor relative to both Normal and Slow CDPR for smoking frequency and the NDSS. Slow CDPR was associated with the highest probability of smoking discontinuation at baseline. However, due to CDPR time trend differences, by young adulthood these baseline effects had been reordered such that the greatest risks for smoking frequency and the NDSS were associated with Normal CDPR. Reduced metabolism CYP2A6 genotypes are associated with both risk and protective effects in novice smokers. However, differences in the time-by-CDPR effects result in a reordering of genotype effects such that normal metabolism becomes the risk variant by young adulthood, as has been reliably reported in older smokers.

The Nicotine Metabolite Ratio in Pregnancy Measured by trans-3'-Hydroxycotinine to Cotinine Ratio: Characteristics and Relationship With Smoking Cessation.

Nicotine replacement therapy (NRT) helps nonpregnant smokers quit, but there is no evidence that standard dose NRT is effective in pregnancy. As nicotine metabolism increases in pregnancy, this could reduce NRT efficacy. Using the ratio of trans-3'-hydroxycotinine to cotinine, the nicotine metabolite ratio (NMR), we investigated relationships between the rate of nicotine metabolism, maternal characteristics and smoking cessation in pregnant women recruited to a randomized controlled trial of NRT. Data from 1,050 pregnant smokers in the Smoking, Nicotine and Pregnancy trial who were of 12-24 weeks gestation had exhaled carbon monoxide readings of ≥8 ppm at recruitment and who were randomized to NRT or placebo patches were used. Linear and logistic regression investigated associations between maternal characteristics and NMR and also between NMR and subsequent validated cessation from smoking. Six hundred and sixty-two women (63%) provided blood samples for NMR estimation. Higher NMR was associated with increased cigarette consumption prior to pregnancy. At 1 month (odds ratio [OR] = 0.87; 95% CI = 0.76-0.99; p = .043) and delivery (OR = 0.79; 95% CI = 0.66-0.95; p = .010), there was a significant negative association between a 0.1 unit increase in NMR and odds of achieving cessation after adjusting for possible confounders. There was no evidence for an interaction between a 0.1 unit increase in NMR and treatment assignment on the odds of cessation at 1 month post-quit date (p = .556). Pregnant women who metabolize nicotine more rapidly are less likely to achieve cessation when they try to quit smoking. There is no evidence that NRT is more effective in women who metabolize nicotine more slowly.

Importance of national context in the translation of personalised treatments for smoking cessation

Caryn Lerman and colleagues1 present the results of a placebo-controlled randomised trial of two smoking cessation treatments, nicotine replacement therapy patch and varenicline, in a sample of adult, daily smokers. Participants were stratified within each treatment group according to a biomarker of nicotine metabolism, the nicotine metabolite ratio (NMR; 3-hydroxycotinine:cotinine). The NMR reflects the rates of nicotine and cotinine metabolism, which are affected primarily by the activity of the CYP2A6 enzyme, which is itself affected by genetic and environmental factors.

Nicotine Dependence, Nicotine Metabolism, and the Extent of Compensation in Response to Reduced Nicotine Content Cigarettes.

The Food and Drug Administration has the authority to regulate tobacco product constituents, including nicotine, to promote public health. Reducing the nicotine content in cigarettes may lead to lower levels of addiction. Smokers however may compensate by smoking more cigarettes and/or smoking more intensely. The objective of this study was to test whether individual differences in the level of nicotine dependence (as measured by the Fagerstrom Test of Cigarette Dependence [FTCD]) and/or the rate of nicotine metabolism influence smoking behavior and exposure to tobacco toxicants when smokers are switched to reduced nicotine content cigarettes (RNC). Data from 51 participants from a previously published clinical trial of RNC were analyzed. Nicotine content of cigarettes was progressively reduced over 6 months and measures of smoking behavior, as well as nicotine metabolites and tobacco smoke toxicant exposure, CYP2A6 and nicotinic CHRNA5-A3-B4 (rs1051730) genotype were measured. Higher baseline FTCD predicted smoking more cigarettes per day (CPD), higher cotinine and smoke toxicant levels while smoking RNC throughout the study, with no interaction by RNC level. Time to first cigarette (TFC) was associated with differences in compensation. TFC within 10 min was associated with a greater increase in CPD compared to TFC greater than 10 min. Neither rate of nicotine metabolism, nor CYP2A6 or nicotinic receptor genotype, had an effect on the outcome variables of interest. FTCD is associated with overall exposure to nicotine and other constituents of tobacco smoke, while a short TFC is associated with an increased compensatory response after switching to RNC.

Comparison of questionnaire responses with biomarkers of tobacco smoke exposure in a Canadian birth cohort at three months of age

Background Exposure to tobacco smoke increases the risk for several adverse health effects in children including wheeze, asthma, and asthma exacerbation. Accurately assessing tobacco smoke exposure is important for understanding and preventing these health effects. Questionnaires are a flexible and relatively inexpensive method of assessing exposure, but biomarkers of tobacco smoke exposure are considered more accurate. We developed questionnaire-based exposure models predicting urinary levels of biomarkers cotinine and trans-3’-hydroxycotinine (3HC) (metabolites of nicotine) in 3-month old infants using parent-reported questionnaire responses about tobacco smoke exposure from the Canadian Healthy Infant Longitudinal Development (CHILD) Study. Methods We used a manual model building process to build multiple linear regression models predicting urinary concentrations of cotinine, 3HC, and the sum of cotinine and 3HC on a molar basis (Cot+3HC) for 987, 1003, and 983 infants, respectively. Questions were included on the infant’s exposure assessed at 3 months of age and tobacco smoke odour in the home. We also included questions on maternal smoking status and history, passive exposure, and family socio-economic status assessed during pregnancy, as potential indirect measures of the infant’s exposure at 3 months. Adjusted R2 values were maximized in the final models. Results During pregnancy, the prevalence of maternal smoking was 2.4 %, and 115 (11.4 %) mothers reported smoking by at least 1 person at home. Of the 144 (14.3 %) infants whose mothers reported that smoking occurred at home when their child was 3 months, 129 (89.6%) and 136 (94.4%) had cotinine and 3HC levels above the detection limit (0.03 ng/mL), respectively. Of the 811 infants who had no parent-reported exposure at 3 months, 538 (66.3%) and 715 (88.2%) had detectable cotinine and 3HC levels, respectively. After correcting for urine dilution, the geometric mean levels were 0.085 ng/mL for cotinine, 0.20 ng/mL for 3HC, and 1.62 picomole/mL for Cot+3HC. The final questionnaire models explained 43.4%, 41.0%, and 42.9% of the variance in cotinine, 3HC, and Cot+3HC levels, respectively. Conclusions Our results indicate that exposure of these infants to tobacco smoke is not completely captured by questionnaires, suggesting that exposure assessment could be improved by using a combination of biomarker and questionnaire methods. Though more detectable, the inclusion of 3HC did not increase the ability of the questionnaires to explain variance in metabolite levels, but 3HC may be important since the ratio of 3HC to cotinine can be used to quantify the rate of nicotine metabolism and variation within populations

Variation in P450 oxidoreductase (POR) A503V and flavin-containing monooxygenase (FMO)-3 E158K is associated with minor alterations in nicotine metabolism, but does not alter cigarette consumption

The rates of nicotine metabolism differ widely, even after controlling for genetic variation in the major nicotine-metabolizing enzyme, CYP2A6. Genetic variants in an additional nicotine-metabolizing enzyme, flavin-containing monooxygenase (FMO)-3, and an obligate microsomal CYP-supportive enzyme, cytochrome P450 oxidoreductase (POR), were investigated. We examined the impact of FMO3 E158K and POR A503V before and after stratifying by CYP2A6 metabolism group. In 130 nonsmokers of African descent who received 4 mg oral nicotine, FMO3 158K trended toward slower nicotine metabolism in reduced CYP2A6 metabolizers (P=0.07) only, whereas POR 503V was associated with faster CYP2A6 activity (nicotine metabolite ratio) in normal (P=0.03), but not reduced, CYP2A6 metabolizers. Neither FMO3 158K nor POR 503V significantly altered the nicotine metabolic ratio (N=659), cigarette consumption (N=667), or urine total nicotine equivalents (N=418) in smokers of African descent. Thus, FMO3 E158K and POR A503V are minor sources of nicotine metabolism variation, insufficient to appreciably alter smoking.

Novel CYP2A6 variants identified in African Americans are associated with slow nicotine metabolism in vitro and in vivo

Nicotine, the main addictive ingredient in tobacco, is metabolically inactivated to cotinine primarily by the hepatic enzyme CYP2A6. Considerable genetic variation in the CYP2A6 gene results in large variation in the rates of nicotine metabolism, which in turn alters smoking behaviours (e.g. amount of cigarettes smoked, risk for dependence and success in smoking cessation). The aim of this study was to identify and characterize novel variants in CYP2A6. The CYP2A6 gene from African American phenotypically slow nicotine metabolizers was sequenced and seven novel variants were identified [CYP2A6*39 (V68M), CYP2A6*40 (I149M), CYP2A6*41 (R265Q), CYP2A6*42 (I268T), CYP2A6*43 (T303I), CYP2A6*44 (E390K), CYP2A6*44 (L462P)]. Variants were introduced into a bi-cistronic cDNA expression construct containing CYP2A6 and P450 oxidoreductase and assessed for protein expression, enzymatic activity and stability as evaluated using western blotting and nicotine metabolism. Genotyping assays were developed and allelic frequencies were assessed in 534 African Americans. The variants showed significantly lower protein expression (P<0.001) when compared with the wild-type as well as reduced metabolism of nicotine to cotinine when controlling for cDNA expression using P450 oxidoreductase (P<0.001). The variants also showed reduced stability at 37°C. Allelic frequencies ranged from 0.1 to 0.6% with a collective genotype frequency of 3.2%; the impact in vitro correlated significantly with in-vivo activity (R(2)=0.40-0.48, P<0.05). Together, those with a novel variant had significantly lower nicotine metabolism in vivo than those without genetic variants (P<0.01). Here, we identified a number of novel variants with reduced/loss of CYP2A6 activity, increasing our understanding of CYP2A6 genetic variability.