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Background: Twin and family studies have demonstrated that adolescent alcohol use and behavior problems are influenced by a combination of genetic and environmental factors. More recently, studies have begun to investigate how genetic and environmental influences may interact, with efforts underway to identify specific environmental variables that moderate the expression of genetic predispositions. Previously, we have reported that community-level factors, including urban/rural residency, migration rates, and prevalence of young adults, moderate the importance of genetic effects on alcohol use in late adolescence (ages 16 to 18). Here, we extend these findings to test for moderating effects of these socioregional factors on alcohol use and behavior problems assessed in a younger sample of adolescent Finnish twins. Methods: Using data from the population-based Finnish twin study, FinnTwin12, biometric twin models were fit to data on >1,400 twin pairs to examine the significance of each of the socioregional moderating variables on alcohol use measured at age 14, and behavior problems, measured at age 12. Results: We find no evidence of a moderating role of these socioregional variables on alcohol use; however, there was significant moderation of genetic influences on behavior problems, with effects limited to girls. Genetic influences assumed greater importance in urban settings, communities with greater migration, and communities with a higher percentage of slightly older adolescents. Conclusions: The moderation effects observed on behavior problems in early adolescence paralleled the effects found on alcohol use late in adolescence in an independent sample, providing further support for the idea that behavior problems may represent an earlier manifestation of the predisposition to subsequent alcohol problems. Our findings also support the growing body of evidence suggesting that females may be more susceptible to a variety of environmental influences than males.

Context: As we identify genes involved in psychiatric disorders, the next step will be to study how the risk associated with susceptibility genes manifests across development and in conjunction with the environment. We describe analyses aimed at characterizing the pathway of risk associated with GABRA2, a gene previously associated with adult alcohol dependence, in a community sample of children followed longitudinally from childhood through young adulthood. Objective: To test for an association between GABRA2 and trajectories of externalizing behavior from adolescence to young adulthood and for moderation of genetic effects by parental monitoring. Design: Data were analyzed from the Child Development Project, with yearly assessments conducted since that time. A saliva sample was collected for DNA at the 2006 follow-up, with a 93% response rate in the target sample. Growth mixture modeling was conducted using Mplus to identify trajectories of externalizing behavior and to test for effects of GABRA2 sequence variants and parental monitoring. Setting: Nashville and Knoxville, Tennessee, and Bloomington, Indiana. Participants: A community-based sample of families enrolled at 3 sites as children entered kindergarten in 1987 and 1988. Analyses for the white subset of the sample (n=378) are reported here. Main Outcome Measures: Parental monitoring measured at 11 years of age; Child Behavior Checklist youth reports of externalizing behavior at ages 12, 14, 15, 16, 17, 19, 20, 21, and 22 years. Results: Two classes of externalizing behavior emerged: a stable high externalizing class and a moderate decreasing externalizing behavior class. TheGABRA2 gene was associated with class membership, with subjects who showed persistent elevated trajectories of externalizing behavior more likely to carry the genotype previously associated with increased risk of adult alcohol dependence. A significant interaction with parental monitoring emerged; the association of GABRA2 with externalizing trajectories diminished with high levels of parental monitoring. Conclusions: These analyses underscore the importance of studying genetic effects across development and of identifying environmental factors that moderate risk.

Objective: The low level of response (LR) to alcohol is an endophenotype related to heavier drinking and alcohol problems. Structural equation models (SEMs) indicate LR affects alcohol outcomes (ALCOUT) both directly and through mediation by drinking in peers (PEER), alcohol expectancies (EXPECT), and drinking to cope with stress (COPE), with some variation depending on the sample tested. This article presents the fi rst full test of this LR-based model in young subjects from the Collaborative Study on the Genetics of Alcoholism (COGA). Method: Data were generated from 325 12- to 22-year-old (47.4% male) drinking offspring from COGA families, using the Self- Report of the Effects of Alcohol questionnaire to determine LR early in the drinking career and a validated, structured interview for demography and alcohol use/problem patterns. Standardized questionnaires were used to measure PEER, EXPECT, and COPE, with the model tested through the maximum likelihood estimation for analyses of the variance/covari- ance matrix using both Amos and Mplus. Results: The SEM yielded good fi t characteristics and explained 59% of the variance, with LR relating both directly to ALCOUT and as partially mediated by PEER and COPE. Although GENDER related to both LR and ALCOUT in the model, and AGE related to ALCOUT, the SEM results were invariant across both AGE and GENDER, with generally similar invariant results regarding the presence or absence of an alcohol-use disorder diagnosis. Conclusions: The results support the applicability of the LR-based model of heavy drinking and alcohol problems in the COGA offspring, a group with different demography compared with the two other samples of adolescents tested to date. The modest differences observed across samples will be evaluated in future research to enhance understanding of how the model operates across socioeconomic groups. (J. Stud. Alcohol Drugs 70: 436-445, 2009)

The abuse of licit and illicit drugs constitutes one of the leading public health problems in the world. There are tens of millions of alcohol or drug abusers and more than a billion smokers in the world (WHO, 2002). Each year, substance abuse results in the loss of tens of millions of dollars due to health-care costs and lost productivity (Cartwright, 2008; Rehm, Taylor, & Room, 2006). Substance abuse also shortens lives, increases risk for chronic disabling illness, and results in untold social costs in terms of broken families, ruined careers, and violent victimization (Goldman, Oroszi, & Ducci, 2005). While effective treatments for addiction exist, the amelioration of the societal burden of substance abuse will require more effective prevention. Behavioral genetic research in this area is part of a larger effort aimed at improving prevention and intervention efforts by bringing about a better understanding of the origins of substance use disorders (O’Brien, 2008). Behavioral genetic research has helped to transform the addictions research field. No longer are substance use disorders considered a form of moral weakness or an outgrowth of an unresolved dynamic conflict. Rather, there is now widespread recognition that substance use disorders have a neurological basis and are genetically influenced. The present chapter provides an overview of the behavioral genetic research that has helped to achieve this transition. This is an extremely active area for behavioral genetic research, especially over the past 10 years, and it is well beyond the scope of the present review to comprehensively cover all relevant research. This review is consequently selective but intended to be representative. The scope of our review includes behavioral genetic research on the use and abuse of both common licit (i.e., alcohol) and illicit

Genomic studies of cannabis use disorders have been limited. The cannabinoid receptor 1 gene (CNR1) on chromosome 6q14-15 is an excellent candidate gene for cannabis dependence due to the important role of the G-protein coupled receptor encoded by this gene in the rewarding effects of Delta9-tetrahydrocannabinol. Previous studies have found equivocal evidence for an association between SNPs in CNR1 and a general vulnerability to substance use disorders. We investigate the association between 9 SNPs spanning CNR1 and cannabis dependence in 1,923 individuals. Two SNPs that were previously associated with cannabis dependence in other studies were also significant with this phenotype in our analyses [rs806368 (P = 0.05) and rs806380 (P = 0.009)]. Haplotype analyses revealed the association to be largely driven by the SNP rs806380. These results suggest a role for the cannabinoid receptor 1 gene in cannabis dependence.

Background: Although use of illicit drugs shows varying degree of heritability, the influence of shared and unique environmental factors predominate among adolescents. We explored factors predicting use of cannabis and other illicit drugs among Finnish adolescent twins. Methods: We used longitudinal data from the FinnTwin 12-17 study with baseline at age 11-12 and follow-up at ages 14 and 171/2, including 4138 individuals. The outcome was self-reported ever use of cannabis or other illicit drugs at age 171/2. The potential predictors were measures reported by the twins, their parents or teachers. As individual factors we tested smoking, alcohol use, behavioral and emotional problems; as peer factors: number of smoking friends and acquaintances with drug experience; as family factors: parental substance use, socio-economic status and pre-natal exposure to nicotine. We used logistic regression models, controlling for twinship, age and sex, to compute odds ratios (OR) for each potential predictor. To adjust for within-family confounds, we conducted conditional logistic regressions among 246 twin pairs discordant for drug use. Results: 13.5% of subjects had initiated use of cannabis or other illicit drugs by age of 171/2. When adjusted for within-family confounds, smoking, drinking, and aggressiveness, as well as smoking and drug use among peers predicted use of illicit drugs. In the final regression model, the significant predictors were female sex, early smoking onset, drinking to intoxication, having smoking peers and acquaintances with drug experience, father's weekly drinking to intoxication, and aggressive behavior among boys. Smoking initiation by age of 12 was the most powerful predictor among individuals (OR = 26, p < 0.001) and within discordant pairs (OR = 22, p < 0.001). Conclusions: Early onset smoking is a powerful predictor for subsequent use of illicit drugs among Finnish adolescents, but the causal nature of this relationship needs to be clarified. (C) 2008 Elsevier Ireland Ltd. All rights reserved.

Objective: Research in animals has shown that exposure to stressors during pregnancy is associated with offspring behavioural disorders. We aimed to study the effect of in utero exposure to the Chernobyl disaster in 1986, and maternal anxiety presumably associated with that exposure, on behaviour disorder observed at age 14. Method: Exposed (n = 232) and non-exposed Finnish twins (n = 572) were compared. A semi-structured interview was used to assess lifetime symptoms of depression, generalized anxiety disorder, attention deficit hyperactivity disorder, conduct disorder and oppositional defiant disorder symptoms. Results: Adolescents who were exposed from the second trimester in pregnancy onwards, had a 2.32-fold risk (95% CI: 1.13-4.72) of having lifetime depression symptoms, an increased risk of fulfilling DSM-III-R criteria of a major depressive disorder (OR = 2.48, 95% CI: 1.06-5.7), and a 2.01-fold risk (95% CI: 1.14-3.52) of having attention deficit hyperactivity disorder symptoms. Conclusion: Perturbations in fetal brain development during the second trimester may be associated with the increased prevalence of depressive and attention deficit hyperactivity disorder symptoms.

Objective: To test whether variation in the gene encoding the enzyme catechol-O-methyltransferase (COMT), which catalyzes the breakdown of dopamine and other catecholamine neurotransmitters, is associated with the risk for alcohol dependence and habitual smoking. Methods: Single nucleotide polymophisms (SNPs) were genotyped in a sample of 219 multiplex alcohol-dependent families of European American descent from the Collaborative Study on the Genetics of Alcoholism (COGA). Family-based tests of association were performed to evaluate the evidence of association between the 18 SNPs distributed throughout COMT, including the functional Val158Met polymorphism, and the phenotypes of alcohol dependence, early onset alcohol dependence, habitual smoking, and comorbid alcohol dependence and habitual smoking. Results: No significant, consistent evidence of association was found with alcohol dependence, early onset alcohol dependence, habitual smoking or the comorbid phenotype. There was no evidence that the functional Val158Met polymorphism, previously reported to be associated with these phenotypes, was associated with any of them. Conclusion: Despite the substantial size of this study, we did not find evidence to support an association between alcohol dependence or habitual smoking and variation in COMT.

Background: There is an extensive and inconsistent literature on the association of the dopamine D2 receptor gene (DRD2) with alcohol dependence. Conflicting results have been attributed to differences in the severity of the alcohol dependence phenotype across studies, failure to exclude related disorders from comparison groups, and artifacts of population-stratification. Recently the genetic polymorphism most widely analyzed in DRD2, Taq1A, has been discovered to reside in a neighboring gene, ankyrin repeat and kinase domain containing 1 (ANKK1), located 10 kb downstream from DRD2. Methods: To more carefully characterize evidence for association across this region, we genotyped 26 single nucleotide polymorphisms (SNPs) spanning DRD2 and ANKK1 in a sample of 219 Caucasian families (n = 1,923) from the Collaborative Study on the Genetics of Alcoholism (COGA), making this the most extensive analysis to date of association between this region and alcohol dependence. We used family-based analyses robust to population-stratification, and we made use of rich phenotypic data to analyze alcohol dependence and subtypes hypothesized in the literature to be more directly influenced by DRD2. Results: We found that the evidence for association is strongest in the 5′ linkage disequilibrium block of ANKK1 (that does not contain Taq1A), with weak evidence of association with a small number of SNPs in DRD2. The association in ANKK1 is strongest among the subsets of alcoholics with medical complications and with antisocial personality disorder. Conclusions: More extensive genotyping across DRD2 and ANKK1 suggests that the association with alcohol dependence observed in this region may be due to genetic variants in the ANKK1 gene. ANKK1 is involved in signal transduction pathways and is a plausible biological candidate for involvement in addictive disorders.

Background: Studies have found that genomic variation in the gene SNCA, which encodes the protein a-synuclein, may contribute to the variation in alcohol consumption in an inbred rat model of alcohol preference. Studies in humans have provided support for an association between SNCA and craving for alcohol. Methods: To examine the role of this gene in alcohol dependence and related phenotypes, 30 single nucleotide polymorphisms (SNPs) were genotyped across the SNCA gene in a sample of 219 multiplex alcoholic families of European American descent. Two phenotypes, alcohol dependence and alcohol craving, were analyzed using the pedigree disequilibrium test. Results: There was no evidence of association between any of the SNCA SNPs and alcohol dependence (p � 0.13). In contrast, 8 SNPs provided evidence of association (po0.05) with the phenotype of alcohol craving. Haplotype analysis further supported evidence of an association with alcohol craving; a haplotype encompassing SNPs in intron 4 through the region downstream of the gene was overtransmitted to cravers and a second haplotype was overtransmitted to noncravers. Conclusions: These results suggest that variation in SNCA contributes to alcohol craving, a common, although not uniform, feature of alcohol dependence.

Background A novel family of G protein-coupled receptors, TAS2Rs, has recently been characterized and linked to sensitivity to bitter taste compounds. We have previously reported that a missense mutation in the TAS2R16 gene reduces the sensitivity of the receptor to bitter-taste stimuli and that it is associated with risk for alcohol dependence. Other family-based studies on the genetic transmittance of taste perception have previously demonstrated a correlation between genetic variation in TAS2R38 and sensitivity to bitter-taste compounds such as phenylthiocarbamide (PTC) and 6-n-propylthiouracil (PROP). Haplotypes resulting from 3 common nonsynonymous coding single-nucleotide polymorphisms in the TAS2R38 gene have been shown to alter receptor functions and taste sensitivity to PTC and PROP. The perceived bitterness of PROP has also been associated with oral sensation and drinking behaviors. Methods We used family-based association methods to test for association between TAS2R38 haplotypes and alcohol dependence as well as a measure of alcohol consumption (Maxdrinks) and age of onset of drinking behaviors in a sample of families densely affected with alcoholism. We have also extended our analysis of TAS2R16 to include the Maxdrinks phenotype. Results A positive correlation was observed between TAS2R38 haplotypes and Maxdrinks in Collaborative Study on the Genetics of Alcoholism (COGA) high-risk women of African-American origin. The common taster haplotype is significantly associated with a lower mean Maxdrinks compared with the other haplotypes. Similarly, the allele of TAS2R16 that is associated with a lower risk for alcohol dependence is also associated with lower mean Maxdrinks scores in African-American families. In contrast to the previously reported significant association between TAS2R16 and alcohol dependence, we found no evidence that TAS2R38 haplotypes influence alcohol dependence in the COGA dataset. Conclusion Functional variants in both TAS2R16 and TAS2R38 correlate with alcohol consumption in African-American families.

s / Drug and Alcohol Dependence 146 (2015) e202–e284 e267 Inhibitory control in patients with cocaine abuse is equally disrupted by generic and personally salient cocaine images Gregory J. DiGirolamo1,2, David Smelson2, Nathan Guevremont2, Gerardo Gonzalez2 1 Psychology, College of the Holy Cross, Worcester, MA, United States 2 Psychiatry, University of Massachusetts, Medical School, Worcester, MA, United States Aims: Todetermine the extent that salient cocaine cues increase breakdown of control in patients with cocaine abuse, wemeasured control over behavior in a task where patients must inhibit an impulsive action and instead look away from a suddenly appearing image. Cueswere personally salient cocaine or neutral stimuli (participants handled them and rated their craving; photos of them touching the cues were used during the task) and generic cocaine (photos of cocaine-related cues) and neutral stimuli. Methods: Twenty-five treatment-seeking patients (M=37.2 yrs; 19males)with a history of cocaine abuse (M=14.6 yrs) and use of cocaine within the last month (16.2 days of use) were inducted into the study. Initial assessments included Voris cocaine craving measures and anti-saccade performance. Results: Voris craving scores were significantly higher (t=5.82, p< .0001) after handling the cocaine cues (M=27.3) than after handling the neutral cues (M=13.3). Inhibitory control was significantly worse (t=2.1, p< .05) for the salient cocaine cues (M=37%) than the salient neutral cues (M=33%). Likewise, errorswere significantly higher (t=2.8, p< .01) for the generic cocaine cues (M=34%) than the generic neutral cues (M=29%). Importantly, there was no hint of a statistical (t

Aims: Motivational interviewing (MI) has been shown to be effective as a short-term therapeutic intervention for alcohol abuse. However, the researchon theeffectivenessofMI forotherdrugpopulations is mixed. The inconsistent findings may, in part, be due to the severity of problems experienced by these populations and the relative brevity of the 1–3 sessions typical of an MI intervention. The current study is a Stage 2 clinical trial that addresses these gaps in the literature by studying a methamphetamine (MA) dependent population and extending MI therapy to 9 sessions. Methods: Participants were randomized to (1) a 9-session intensive MI intervention (n=111) or (2) one session of standard MI with a time equivalence activity (n=106). All participants were followed at 2, 4, and 6months. Data collection included days of MA use, ASI drug, and ASI psychiatric severity. Results: Longitudinal analyses were estimated via the use of random intercept models assessing averaged treatment effects from baseline. Both conditions had significant decreases in MA use and ASI drug scores though significant differences between MI conditions were not observed. However, ASI psychiatric severity scores significantly declined for the intensive MI condition but not for the standard MI condition. Further examination of psychiatric symptoms showed significant decreases in the number of days of psychological problems compared to the standardMI condition and significant decreases in the number of days of depression for the intensive MI condition. Conclusions: This clinical trial focused on MA dependent individuals, a population that frequently presentswith serious psychiatric symptomatology at treatment entry, thereby complicating treatment efforts. Our findings demonstrate that standard MI may be equally beneficial in helpingMA clients reduce theirMA use and problem severity but intensive MI may help alleviate psychological problems not adequately addressed in shorterMI interventions. Future study should include long-term follow-up and examine the elements of MI therapy that may work as moderators to successful recovery. Financial support: NIDA R01 DA024714.

The cholinergic neurotransmitter system is thought to be involved in many aspects of memory, attention, and higher cognition. In the Collaborative Study on the Genetics of Alcoholism (COGA) sample, we have previously reported linkage and association to the cholinergic muscarinic 2 receptor gene (CHRM2) on chromosome 7 with evoked EEG oscillations (Jones et al. 2004), providing evidence that this gene may be involved in human brain dynamics and cognition. In addition, a small number of genetic markers were genotyped in CHRM2 in the Minnesota Twin and Family Study (Comings et al. 2003) and a Dutch family study (Gosso et al. 2006, in press) and both research groups found evidence that this gene may be involved in intelligence. In the COGA sample, we have extensively genotyped SNPs within and flanking the CHRM2 gene. We find evidence of association with multiple SNPs across CHRM2 and Performance IQ, as measured by the Wechsler Adult Intelligence Scale-Revised (WAIS-R). These results remain significant after taking into account alcohol dependence and depression diagnoses in the sample.

Background: A wealth of literature supports the role of γ-aminobutyric acid (GABA) in neurobiological pathways contributing to alcohol dependence and related phenotypes. Animal studies have consistently tied rodent homologs of the GABAA receptor genes on human chromosome 5q to alcohol-related behaviors; however, human studies have produced mixed results. Family-based association analyses previously conducted in the Collaborative Study on the Genetics of Alcoholism (COGA) sample yielded no evidence of association with Diagnostic and Statistical Manual of Mental Disorder—fourth edition (DSM-IV) alcohol dependence and these genes. As a follow-up to that study, we examined several alcohol-related behaviors in the COGA sample as follows: (1) a broader definition of alcohol dependence, including DSM-III-R symptoms and Feighner criteria (referred to as COGA alcohol dependence); (2) withdrawal; (3) history of alcohol-induced blackouts; (4) level of response to alcohol; (5) age of onset of regular drinking; and (6) age at first drunkenness. Methods: Family-based association tests were conducted, using multiple single-nucleotide polymorphisms (SNPs) in each of the 4 GABAA receptor genes on chromosome 5q. Results: In GABRA1, we found evidence of association with several of the drinking behavior phenotypes, including COGA alcohol dependence, history of blackouts, age at first drunkenness, and level of response to alcohol. We did not find consistent evidence of association with the remaining genes and any of the phenotypes. Conclusions: We found evidence for association between GABRA1 and COGA alcohol dependence, history of blackouts, age at first drunkenness, and level of response to alcohol. These analyses suggest that efforts to characterize genetic contributions to alcohol dependence may benefit by examining alcohol-related behaviors in addition to clinical alcohol dependence diagnoses.

Linkage evidence indicated that gene(s) located on chromosome 4q, in the region of the alcohol dehydrogenase (ADH) genes, affected risk for alcoholism. We genotyped 110 single nucleotide polymorphisms (SNPs) across the seven ADH genes and analyzed their association with alcoholism in a set of families with multiple alcoholic members, using the pedigree disequilibrium test. There was strong evidence that variations in ADH4 are associated with alcoholism: 12 SNPs were significantly associated. The region of strongest association ran from intron 1 to 19.5 kb beyond the 3 0 end of the gene. Haplotype tag SNPs were selected for the block in the ADH4 gene that provided evidence of association and subsequently used in association analysis; the haplotype was significantly associated with alcoholism (P 5 0.01) There was weaker evidence that variations in ADH1A and ADH1B might also play a role in modifying risk. Among African-Americans, there was evidence that the ADH1B*3 allele was protective.

Background Depressions that fail to meet the diagnostic criteria for major depressive disorder (MDD) may be underdiagnosed and undertreated in adolescent population. Traditionally, they are not considered as serious conditions and the phenomenological nature and clinical correlates of these disorders are largely unknown. In the present study, we used a large, representative and age-standardized sample of adolescents to examine the phenomenology and clinical correlates of minor depression, a poorly understood condition included in the category of Depressive Disorder Not Otherwise Specified in Diagnostic and Statistical Manual of Mental Disorders–Fourth Edition–Text Revised (DSM-IV-TR). Methods 909 girls and 945 boys, with mean age of 14, were interviewed by professionals using the Semi-Structured Assessment for the Genetics of Alcoholism (SSAGA). Results Although clearly milder condition than MDD, minor depression was associated with marked suicidal thoughts, plans and attempts, recurrences and a high degree of comorbidity. At this early age, despite that 14% of adolescents under 15 had suffered from depressive conditions with severe clinical implications, most of them failing to meet the diagnostic threshold for MDD, only 1.7% had received any psychiatric treatment. 40% of depressive adolescents who had attempted suicide had no contact with mental health services. Limitations Analyzed in a cross-sectional setting, no conclusions about long-term implications could be made. Conclusions The results highlight the clinical and public health significance of non-MDD depressions, e.g. minor depression, which need to be more carefully identified and treated at early age.

Genetic and environmental effects are not independent sources of influence on behavior, but rather, genotype can influence an individual's exposure to certain environments, how that individual experiences the environment, and the degree of influence that certain environments exert. This phenomenon is called gene-environment correlation. Three types of gene-environment correlation are discussed: passive, evocative, and active. Examples of each type of gene-environment correlation, and support for these effects from the literature are presented. Keywords: gene-environment correlation; passive; active; evocative

The progression to alcohol dependence unfolds across multiple stages, including the decision to initiate use, the development of regular patterns of use, and (for some individuals) the subsequent development of problems associated with alcohol use. Using data from two population-based, longitudinal twin studies, FinnTwin16 (FT16) and FinnTwin12 (FT12), we applied multiple stage genetic models (Heath et al., Twin Res. 5 (2002) 113) to better understand the extent to which genetic and environmental influences impact the initiation of alcohol use, frequency of use in adolescence and young adulthood, and alcohol problems in young adulthood. Shared environmental factors played a large role in initiation, and a more moderate role on frequency of use, and it was largely the same influences acting across these stages of use. However, there was no significant evidence of shared environmental influences on alcohol problems in early adulthood. Problems were largely influenced by genetic factors that overlapped with genetic influences on frequency of use. Unique environmental factors were largely specific to each stage, with some overlap between alcohol problems and frequency of use at age 25.

Conduct disorder (CD), attention deficit hyperactivity disorder (ADHD), and oppositional defiant disorder (ODD) are common childhood externalizing disorders that frequently co-occur. However, the causes of their comorbidity are not well understood. To address that question, we analyzed data from >600 Finnish twin pairs, who completed standardized interviews at age 14. Behavior genetic methods were used to examine how genetic/environmental factors contribute to each disorder's symptoms and to their covariation. We found significant genetic effects on each disorder with only modest evidence of shared environmental influences. Our data suggest the comorbidity among CD, ADHD, and ODD is primarily explained by shared genetic influences; however, each disorder was also under unique genetic influence, supporting the distinction of each disorder.

The use of endophenotypes has been proposed as a strategy to aid gene identification efforts for complex phenotypes [Gottesman, I. I., and Shields J. (1972). Schizophrenia and Genetics: A Twin Study Vantage Point. London: Academic]. As part of the Collaborative Study of the Genetics of Alcoholism (COGA) project, we have analyzed electrophysiological endophenotypes, in addition to clinical diagnoses, as part of our effort to identify genes involved in the predisposition to alcohol dependence. In this paper we summarize published results from linkage and association analyses of two chromosomal regions in which the use of endophenotypes has successfully led to the identification of genes associated with alcohol dependence [GABRA2 (Edenberg et al., (2004). Am. J. Hum. Genet. 74:705-714) and CHRM2 (Wang et al., (2004). Hum. Mol. Genet. 13:1903-1911)]. Our experience in the COGA project has been that the analysis of endophenotypes provides several advantages over diagnostic phenotypes, including the strength and localization of the linkage signal. Our results provide an illustration of the successful use of endophenotypes to identify genes involved in the predisposition to a complex psychiatric phenotype, a strategy originally proposed by Gottesman and Shields in 1972.

Background: Alcohol misuse and conduct disorder (CD) are strongly associated in adolescents. Is their association due to shared genetic and environmental liabilities? We addressed that question with data obtained from structured interviews of 14-year-old Finnish twins. Methods: A total of 1854 twins completed face-to-face interviews. Univariate models, allowing for sex limitation, were fit to symptom counts for Diagnostic and Statistical Manual of Mental Disorders, Third Edition, Revised diagnoses of CD and alcoholism to examine their genetic and environmental influences. Then, bivariate models evaluated the extent to which genetic and environmental influences contributed to the covariation between symptom counts for the two disorders. Results: A total of 822 twins (44% of the sample) reported one or more symptoms of CD; alcohol dependence symptoms were much less common, reported by only 12%. The correlation between the two symptom counts was 0.50. Models fit to the twin data demonstrated that CD symptoms were under significant genetic influence in both boys and girls, although those influences were predominantly sex specific. In contrast, alcohol dependence symptoms were, at this age, under sex-specific effects of common environments, with no evidence of genetic influences. Accordingly, the substantial covariation between alcohol dependence symptoms and those of CD was attributed entirely to shared environmental effects. Conclusions: At age 14, genetic influences on alcohol dependence symptoms are negligible, and the correlated liabilities between these symptoms and those of CD are to be found in environmental factors that are common to both.

Several correlated phenotypes, alcohol dependence, major depressive syndrome, and an endophenotype of electrophysiological measurements, event-related oscillations (EROs), have demonstrated linkage on the long arm of chromosome 7. Recently, we reported both linkage and association between polymorphisms in the gene encoding the muscarinic acetylcholine receptor M2 (CHRM2) and EROs. In this study, we evaluated whether genetic variation in the CHRM2 gene is also a risk factor for the correlated clinical characteristics of alcoholism and depression. The CHRM2 gene contains a single coding exon and a large 5' untranslated region encoded by multiple exons that can be alternatively spliced. Families were recruited through an alcohol dependent proband, and multiplex pedigrees were selected for genetic analyses. We examined 11 single nucleotide polymorphisms (SNPs) spanning the CHRM2 gene in these families. Using the UNPHASED pedigree disequilibrium test (PDTPHASE), three SNPs (one in intron 4 and two in intron 5) showed highly significant association with alcoholism (P=0.004-0.007). Two SNPs (both in intron 4) were significantly associated with major depressive syndrome (P=0.004 and 0.017). Haplotype analyses revealed that the most common haplotype (>40% frequency), T-T-T (rs1824024-rs2061174-rs324650), was under-transmitted to affected individuals with alcohol dependence and major depressive syndrome. Different complementary haplotypes were over-transmitted in alcohol dependent and depressed individuals. These findings provide strong evidence that variants within or close to the CHRM2 locus influence risk for two common psychiatric disorders.

Background: Suicide and suicidal behavior are prevalent among individuals with psychiatric disorders, including alcohol dependence. A genome screen was performed in multiplex alcohol dependent families ascertained as part of the Collaborative Study on the Genetics of Alcoholism to identify chromosomal regions of interest related to two types of suicidal behavior: suicide attempts and suicidality. Methods: Sibling pair analyses were used to conduct linkage analyses using both qualitative and quantitative suicide-related phenotypes. The qualitative trait of “suicide attempts” was examined using 59 affected sibling pairs. The quantitative trait of “suicidality” was examined using all possible 1366 sibling pairs and 705 independent sibling pairs. Results: For the qualitative phenotype suicide attempts, chromosome 2 yielded a maximum lod score of 4.2. For the quantitative suicidality index, a maximum lod score of 1.8 was observed on chromosome 3, and a lod score of 1.5 was found on chromosome 1. Conclusions: This study represents the first genome-wide scan of suicidal behavior. Significant evidence of linkage was found on chromosome 2 for the phenotype suicide attempts, the same chromosomal region previously reported to be linked to alcohol dependence in this sample. This finding does not seem to be due solely to an association between suicide and alcohol dependence. There was more modest evidence of linkage to chromosomes 1 and 3 for suicidality; however, these findings did not reach statistical significance. There was no overlap in findings for these two phenotypes of suicidal behavior.

While behavioral genetic studies have suggested that childhood conduct disorder is under genetic influence, studies aimed at gene identification are lacking. This study represents the first genome-wide linkage analysis directed toward identifying genes contributing to conduct disorder. Genome screens of retrospectively reported childhood conduct disorder and conduct disorder symptomatology were carried out in the genetically informative adult sample collected as part of the Collaborative Study on the Genetics of Alcoholism (COGA). The results suggest that regions on chromosomes 19 and 2 may contain genes conferring risk to conduct disorder. Interestingly, the same region on chromosome 2 has also been linked to alcohol dependence in this sample. Childhood conduct disorder is known to be associated with the susceptibility for future alcohol problems. Taken together, these findings suggest that some of the genes contributing to alcohol dependence in adulthood may also contribute to conduct disorder in childhood.

We have reported genetic linkage between bipolar disorder and markers on chromosome 6q16.3-22.1 in the National Institute of Mental Health Genetics Initiative wave 3 pedigrees. Here we test for: 1) robustness of the linkage to differing analysis methods, genotyping error, and gender-specific maps; 2) parent-of-origin effects; and 3) interaction with markers within the schizophrenia linkage region on chromosome 6p.Members of 245 families ascertained through a sibling pair affected with bipolar I or schizoaffective-bipolar disorder were genotyped with 18 markers spanning chromosome 6. Nonparametric linkage analysis was performed.Linkage to 6q is robust to analysis method, gender-specific map differences, and genotyping error. The locus confers a 1.4-fold increased risk. Affected siblings share the maternal more often than the paternal chromosome (p =.006), which could reflect a maternal parent-of-origin effect. There is a positive correlation between family-specific linkage scores on 6q and those on 6p22.2 (r =.26; p.0001). Linkage analysis for each locus conditioned on evidence of linkage to the other increases the evidence for linkage at both loci (p.0005). Logarithm of the odds (LOD) scores increased from 2.26 to 5.42 on 6q and from.35 to 2.26 on 6p22.2.These results support linkage of bipolar disorder to 6q, uncover a maternal parent-of-origin effect, and demonstrate an interaction of this locus with one on chromosome 6p22.2, previously linked only to schizophrenia.

In 1989 the National Institute of Mental Health began a collaborative effort to identify genes for bipolar disorder. The first 97 pedigrees showed evidence of linkage to chromosomes 1, 6, 7, 10, 16, and 22 (Nurnberger et al 1997). An additional 56 bipolar families have been genotyped, and the combined sample of 153 pedigrees studied.Three hierarchical affection status models were analyzed with 513 simple sequence repeat markers; 298 were common across all pedigrees. The primary analysis was a nonparametric genome-wide scan. We performed conditional analyses based on epistasis or heterogeneity for five regions.One region, on 16p13, was significant at the genome-wide p.05 level. Four additional chromosomal regions (20p12, 11p15, 6q24, and 10p12) showed nominally significant linkage findings (p/=.01). Conditional analysis assuming epistasis identified a significant increase in linkage at four regions. Families linked to 6q24 showed a significant increase in nonparametric logarithms of the odds (NPL) scores at 5q11 and 7q21. Epistasis also was observed between 20p12 and 13q21, and 16p13 and 9q21.The findings are presented in rank order of nominal significance. Several of these regions have been previously implicated in independent studies of either bipolar disorder or schizophrenia. The strongest finding is at 16p13 at D16S748 with an NPL of 3.3, there is evidence of epistasis between this locus and 9q21. Application of conditional analyses is potentially useful in larger sample collections to identify susceptibility genes of modest influence that may not be identified in a genome-wide scan aimed to identify single gene effects.

Family characteristics influence children's behavioral development, but so do variations in schools, neighborhoods, and communities. We documented extrafamilial environmental effects by fitting maximum likelihood models to questionnaire data collected from double dyads consisting of twins and their classmate controls. The classmate controls in each double dyad were genetic strangers living in separate households, but they shared school, neighborhood, and community environments with their yoked twin pair and with one another. At ages 11 to 12, the control classmates showed significant similarities in religious practices and smoking and drinking patterns, demonstrating that environmental influences outside the family affect children's behavioral development. Familial self-selection of residential neighborhoods may have contributed to these results, but direct effects of variation across communities, neighborhoods, and schools cannot be dismissed, and such effects warrant further study.

We conducted genomewide linkage analyses on 1,152 individuals from 250 families segregating for bipolar disorder and related affective illnesses. These pedigrees were ascertained at 10 sites in the United States, through a proband with bipolar I affective disorder and a sibling with bipolar I or schizoaffective disorder, bipolar type. Uniform methods of ascertainment and assessment were used at all sites. A 9-cM screen was performed by use of 391 markers, with an average heterozygosity of 0.76. Multipoint, nonparametric linkage analyses were conducted in affected relative pairs. Additionally, simulation analyses were performed to determine genomewide significance levels for this study. Three hierarchical models of affection were analyzed. Significant evidence for linkage (genomewide P

Alcohol dependence is a clinically and etiologically heterogeneous disorder. Accordingly, a variety of subtypes of alcohol-dependent individuals have been proposed, and multiple operational definitions of alcohol use, abuse, and dependence have been used in linkage analyses directed toward detecting genes involved in alcohol use and problems. Here, we develop quantitative phenotypes that characterize drinking patterns among both alcoholic and nonalcoholic subjects, and use these phenotypes in subsequent linkage analyses.More than 9000 individuals from alcoholic and control families were administered a semistructured interview and personality questionnaire as part of the initial stage of the Collaborative Study on the Genetics of Alcoholism (COGA). A principal component analysis was conducted on items that captured many of the dimensions of drinking and related behaviors, including aspects of alcohol use, antisocial behavior and affective disturbance when drinking, and personality. Factor scores were computed for all individuals. Nonparametric linkage analyses were conducted on these factor scores, in the initial COGA sample consisting of 987 individuals from 105 extended families, and in a replication sample consisting of 1295 individuals from 157 extended families.Three factors were identified, accounting for 68% of the total variance. The most promising regions of linkage appeared for factor 2, on which higher scores indicate a later age of onset of regular drinking and higher harm avoidance. Chromosome 1 yielded consistent evidence of linkage in both samples, with a maximum lod score of 3.3 when the samples were combined for analysis. Consistent suggestion of linkage also was found to chromosome 15.Developing novel phenotypes that more accurately model the effect of influential genes may help efforts to detect genes involved in complex disorders. Applying principal component analysis in the COGA sample provided support for some regions of linkage previously reported in COGA, and identified other new, promising regions of linkage.

Nuclear magnetic resonance assays allow for measurement of a wide range of metabolic phenotypes. We report here the results of a GWAS on 8,330 Finnish individuals genotyped and imputed at 7.7 million SNPs for a range of 216 serum metabolic phenotypes assessed by NMR of serum samples. We identified significant associations (P < 2.31 - 10 g-10) at 31 loci, including 11 for which there have not been previous reports of associations to a metabolic trait or disorder. Analyses of Finnish twin pairs suggested that the metabolic measures reported here show higher heritability than comparable conventional metabolic phenotypes. In accordance with our expectations, SNPs at the 31 loci associated with individual metabolites account for a greater proportion of the genetic component of trait variance (up to 40%) than is typically observed for conventional serum metabolic phenotypes. The identification of such associations may provide substantial insight into cardiometabolic disorders. © 2012 Nature America, Inc. All rights reserved.

Alcoholism is a complex disease with both genetic and environmental risk factors. To identify genes that affect the risk for alcoholism, we systematically ascertained and carefully assessed individuals in families with multiple alcoholics. Linkage and association analyses suggested that a region of chromosome 4p contained genes affecting a quantitative endophenotype, brain oscillations in the beta frequency range (13-28 Hz), and the risk for alcoholism. To identify the individual genes that affect these phenotypes, we performed linkage disequilibrium analyses of 69 single-nucleotide polymorphism (SNPs) within a cluster of four GABA(A) receptor genes, GABRG1, GABRA2, GABRA4, and GABRB1, at the center of the linked region. GABA(A) receptors mediate important effects of alcohol and also modulate beta frequencies. Thirty-one SNPs in GABRA2, but only 1 of the 20 SNPs in the flanking genes, showed significant association with alcoholism. Twenty-five of the GABRA2 SNPs, but only one of the SNPs in the flanking genes, were associated with the brain oscillations in the beta frequency. The region of strongest association with alcohol dependence extended from intron 3 past the 3' end of GABRA2; all 43 of the consecutive three-SNP haplotypes in this region of GABRA2 were highly significant. A three-SNP haplotype was associated with alcoholism, with P=.000000022. No coding differences were found between the high-risk and low-risk haplotypes, suggesting that the effect is mediated through gene regulation. The very strong association of GABRA2 with both alcohol dependence and the beta frequency of the electroencephalogram, combined with biological evidence for a role of this gene in both phenotypes, suggest that GABRA2 might influence susceptibility to alcohol dependence by modulating the level of neural excitation.

We conducted a genomewide linkage screen of a simple heavy-smoking quantitative trait, the maximum number of cigarettes smoked in a 24-h period, using two independent samples: 289 Australian and 155 Finnish nuclear multiplex families, all of which were of European ancestry and were targeted for DNA analysis by use of probands with a heavy-smoking phenotype. We analyzed the trait, using a regression of identity-by-descent allele sharing on the sum and difference of the trait values for relative pairs. Suggestive linkage was detected on chromosome 22 at 27-29 cM in each sample, with a LOD score of 5.98 at 26.96 cM in the combined sample. After additional markers were used to localize the signal, the LOD score was 5.21 at 25.46 cM. To assess the statistical significance of the LOD score in the combined sample, 1,000 simulated genomewide screens were conducted, resulting in an empirical P value of .006 for the LOD score of 5.21. This linkage signal is driven mainly by the microsatellite marker D22S315 (22.59 cM), which had a single-point LOD score of 5.41 in the combined sample and an empirical P value

A lower level of education often co-occurs with alcohol problems, but factors underlying this co-occurrence are not well understood. Specifically, whether these outcomes share part of their underlying genetic influences has not been widely studied. Educational level also reflects various environmental influences that may moderate the genetic etiology of alcohol problems, but gene-environment interactions between educational attainment and alcohol problems are unknown.We studied the two non-mutually exclusive possibilities of common genetic influences and gene-environment interaction between alcohol problems and low education using a population-based sample (n = 4,858) of Finnish young adult twins (M(age) = 24.5 years, range: 22.8-28.6 years). Alcohol problems were assessed with the Rutgers Alcohol Problem Index and self-reported maximum number of drinks consumed in a 24-hour period. Years of education, based on completed and ongoing studies, represented educational level.Educational level was inversely associated with alcohol problems in young adulthood, and this association was most parsimoniously explained by overlapping genetic influences. Independent of this co-occurrence, higher education was associated with increased relative importance of genetic influences on alcohol problems, whereas environmental factors had a greater effect among twins with lower education.Our findings suggest a complex relationship between educational level and alcohol problems in young adulthood. Lower education is related to higher levels of alcohol problems, and this co-occurrence is influenced by genetic factors affecting both phenotypes. In addition, educational level moderates the importance of genetic and environmental influences on alcohol problems, possibly reflecting differences in social-control mechanisms related to educational level.

Alcohol use and internalizing problems are often positively associated during adolescence and adulthood. However, the basis of this relationship remains poorly understood, and longitudinal data collected in population-based samples could improve the development of etiological models.Using a prospective population-based U.K. cohort, the current study examined the relationship between frequency of drinking during adolescence (ages 13-15, N = 7,100) with problems with depression and anxiety at average age 17 years 10 months (n = 4,292). Analyses were conducted separately by sex and adjusted by the inclusion of potential individual- and familial-level confounders.Among boys, drinking frequency was positively associated with later depression but not anxiety. This association was robust to adjustment for covariates/confounders. Among girls, drinking frequency was related to later depression and anxiety in univariable analyses. In multivariable analyses, only the association with depression remained after adjustment for covariates/confounders. RESULTS were comparable across sexes, although the effect size of drinking frequency was higher among boys.Higher adolescent alcohol use, even at sub-clinical levels, is associated with an increased risk of later problems with depression but may not be associated with an aggregate measure of anxiety. Future research should consider the possibility of differential relationships between multiple measures of adolescent alcohol use and distinct internalizing outcomes later in development.