Journal of Substance Abuse Treatment 38 (2010) 178 – 190

Regular article

Genetic, personality, and environmental predictors of drug use

in adolescents
Bradley T. Conner, (Ph.D.) a , Gerhard S. Hellemann, (Ph.D.) b ,
Terry L. Ritchie, (Ph.D.) c , Ernest P. Noble, (Ph.D., M.D.) c,⁎
a
Department of Psychology, Temple University, Philadelphia, PA 19122, USA
b
Biostatistics Core, Semel Institute for Neuroscience & Human Behavior, David Geffen School of Medicine, University of California Los Angeles,
Los Angeles, CA 90095, USA
c
Alcohol Research Center, Department of Psychiatry & Biobehavioral Sciences, Semel Institute for Neuroscience & Human Behavior,
David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA 90095, USA
Received 3 February 2009; received in revised form 23 June 2009; accepted 21 July 2009

Abstract

During adolescence there is a significant increase in risk-taking behavior, including experimenting with alcohol and drugs, which can lead
to drug dependence. A new hypothesis regarding the genetic mechanisms that lead to drug use is tested using adolescent Caucasian children
of alcoholics (57 males, 54 females; mean age = 14.5 years) data. Variables included in the study were dopaminergic genes (ANKK1 TaqI A,
DRD2 C957T, DRD4 7R, COMT Val/Met substitution, and SLC6A3 9R) and a GABAergic gene (GABRB3), all combinations of genes, a
count of the number of hypodopaminergic genotypes, personality traits, neurocognitive factors, depressive symptoms, and environmental
factors. Separate predictive models were found for males and females. Hypodopaminergic functioning predicted drug use in males; however,
in females, a deleterious environment was the salient predictor. This preliminary study suggests that it is possible to identify children at risk
for problematic drug use prior to the onset of drug dependence. © 2010 Elsevier Inc. All rights reserved.
Keywords: Dopamine genes; GABA genes; Hypodopaminergic functioning; Drug use; Recursive partitioning; Decision tree analysis

1. Introduction understanding of the role genetics plays in the propensity

As children enter the critical developmental period of
adolescence, there is a significant increase in their risk-taking
behavior, including experimenting with alcohol and drugs
(Arnett, 1992; Spear, 2000; Steinberg, 2008). These
behaviors often lead to devastating outcomes, including
drug abuse and dependence (Chen, Storr, & Anthony, 2009;
Grant & Dawson, 1998). These negative outcomes have the
potential to impact adolescents for the rest of their lives.
Although there has been considerable research on adolescent
risk-taking behavior, there are still many gaps in our
⁎ Corresponding author. UCLA Alcohol Research Center, 760 West-
wood Plaza, Los Angeles, CA, USA. Tel.: +1 310 825 1891; fax: +1 310 206
7309.
E-mail address: epnoble@ucla.edu (E.P. Noble).
to engage in risk-taking behavior. Behavioral genetic
research indicates that as much as 40% to 60% of the
vulnerability to develop dependence on substances is
heritable (Button et al., 2006; Han, McGue, & Iacono,
1999). Identification of the specific genetic mechanisms of
this transmission would be invaluable in the prevention and
treatment of these disorders.
Use of most drugs of abuse, including alcohol, is
associated with release of dopamine in the mesocorticolim-
bic system or “reward” pathway of the brain (Di Chiara &
Imperato, 1988). Activation of this dopamine system induces
feelings of reward and pleasure (Bressan & Crippa, 2005;
Spanagel & Weiss, 1999). However, reduced activity of the
dopamine system (hypodopaminergic functioning) can
trigger drug-seeking behavior (Bowirrat & Oscar-Berman,
2005; Melis, Spiga, & Diana, 2005). Variant alleles can

0740-5472/09/$ – see front matter © 2010 Elsevier Inc. All rights reserved.
doi:10.1016/j.jsat.2009.07.004
 B.T. Conner et al. / Journal of Substance Abuse Treatment 38 (2010) 178–190
cause hypodopaminergic functioning through reduced do-
pamine receptor density, blunted response to dopamine, or
enhanced dopamine clearance or metabolism in the reward
pathway (Lachman, 2006). Cessation of chronic drug use
induces a hypodopaminergic state that prompts drug-seeking
behavior in an attempt to address the withdrawal-induced
state (Melis et al., 2005).
These observations are the basis for the development of a
functional hypothesis of drug-seeking and drug use. That
hypothesis is that the presence of a hypodopaminergic state,
regardless of its source, is a primary cause of drug-seeking
behavior. Thus, genetic polymorphisms that cause hypodo-
paminergic functioning may be the causal mechanism of a
genetic predisposition to drug use. This hypothesis is unique
in that it attempts to move the field away from studying the
relationships of genetic polymorphisms to drug use as
associations and, instead, suggests the mechanism that
explains these associations.
The D2 dopamine receptor (DRD2) gene has probably
received the most attention as a candidate for the genetic
transmission of drug use disorders (see Noble, 2003 for
review). The TaqI A is a single nucleotide polymorphism
(SNP rs: 1800497) originally thought to be located at the 3′
region of the DRD2 but which was later found to be located
within exon 8 of an adjacent gene, the ankyrin repeat and
kinase domain containing 1 (ANKK1; Neville, Johnstone, &
Walton, 2004). Neville et al. indicate that the mislocation of
the TaqI A may be attributable to the ANKK1 and the DRD2
being on the same haplotype or the ANKK1 being involved in
reward processes through a signal transduction pathway. The
TaqI A is expressed as two alleles: the A1 and the A2 (Grandy
et al., 1989). Presence of the A1+ genotype (A1/A1, A1/A2),
compared to the A1− genotype (A2/A2), is associated with
reduced DRD2 density (Jönsson et al., 1999; Noble, Blum,
Ritchie, Montgomery, & Sheridan, 1991; Pohjalainen, Rinne,
Någren, Lehikoinen, et al., 1998). This reduction causes
hypodopaminergic functioning in the dopamine reward
pathway. In addition, research supports a predictive relation-
ship from the A1+ genotype to drug use disorders (see Noble,
2003 for review). However, not all studies have supported
this relationship (see Noble, 2003 for review).
Another polymorphism that has received recent attention
is the C957T, a SNP (rs: 6277) at exon 7 of the DRD2 gene
(Duan et al., 2003). This SNP results in the presence of two
alleles: the 957T and the C957. Compared to the T− genotype
(C/C), the T+ genotype (T/T, T/C) is associated with
decreased translation of DRD2 mRNA and diminished
DRD2 mRNA (Duan et al., 2003), leading to decreased
DRD2 density (Hirvonen et al., 2004). This results in
hypodopaminergic functioning, which renders the T +
genotype a good candidate for the transmission of drug use
disorders. In accord with this hypothesis, a recent study has
shown a predictive relationship between the T+ genotype and
alcohol dependence (Hill et al., 2008).
There is evidence that the length of the D4 dopamine
receptor (DRD4) exon 3 variable number of tandem repeats
179
(VNTR) affects DRD4 functioning by modulating the
expression and efficiency of maturation of this receptor
(Schoots & Van Tol, 2003). The 7 repeat (7R) VNTR
requires significantly higher levels of dopamine to produce a
response of the same magnitude as other size VNTRs (Oak,
Oldenhof, & Van Tol, 2000). This reduced sensitivity to
dopamine leads to hypodopaminergic functioning. The 7R
VNTR has been associated with drug use disorders
(McGeary, Esposito-Smythers, Spirito, & Monti, 2007;
Oak et al., 2000). However, not all studies support this
relationship (Lusher, Chandler, & Ball, 2001).
The dopamine transporter protein regulates dopamine-
mediated neurotransmission by rapidly accumulating dopa-
mine that has been released into the synapse (Vandenbergh
et al., 1992). The dopamine transporter gene (SLC6A3 or
DAT1) is localized to chromosome 5p15.3. Within the 3′
noncoding region of DAT1 lies a VNTR polymorphism
(Vandenbergh et al., 1992). The DAT1 9 repeat (9R) VNTR
has been shown to affect gene expression and to enhance
transcription of the dopamine transporter protein (Michel-
haugh, Fiskerstrand, Lovejoy, Bannon, & Quinn, 2001).
This results in enhanced clearance of synaptic dopamine,
leaving reduced amounts of dopamine to activate the
postsynaptic neuron, leading to hypodopaminergic func-
tioning. Presence of the 9R VNTR has been associated with
drug use disorders (Guindalini et al., 2006; Köhnke et al.,
2005); however, not all studies support this relationship
(Vandenbergh et al., 2002).
Catechol-O-methyltransferase (COMT) is an enzyme
involved in the metabolism of dopamine (Lachman et al.,
1996). The 108/158 codon results in a valine (Val) to
methionine (Met) substitution, which reduces COMT
thermostability, even at physiological temperatures, and
significantly decreases enzymatic activity (Lachman et al.,
1996). The Val/Val homozygote is four times more active in
metabolizing dopamine than the Met/Met homozygote
(Lachman et al., 1996). Faster metabolism results in reduced
dopamine availability at the synapse, which diminishes
postsynaptic activation, causing hypodopaminergic func-
tioning. Some studies have shown an association between
the Val variant and drug use disorders (Vandenbergh,
Rodriguez, Miller, Uhl, & Lachman, 1997; Wang et al.,
2001); however, not all studies support this association
(Samochowiec et al., 2006).
Gamma-aminobutyric acid (GABA) receptor genes have
also received some attention as candidates for drug use
disorders. One reason for this is that the dopamine and
GABA systems are functionally interrelated (White, 1996).
Research suggests that dopamine neurons projecting from
the anterior ventral tegmental area to the nucleus accum-
bens are tonically inhibited by GABA through its actions at
the GABAA receptor (Ikemoto, Kohl, & McBride, 1997).
Moreover, it has been shown that alcohol (Theile,
Morikawa, Gonzales, & Morrisett, 2008) or opioid
(Johnson & North, 1992) enhancement of GABAergic
(through GABAA receptor) transmission inhibits the release
180 B.T. Conner et al. / Journal of Substance Abuse Treatment 38 (2010) 178–190
of dopamine in the mesocorticolimbic system. Thus, a
hyperactive GABA system, by inhibiting dopamine release,
could also lead to hypodopaminergic functioning. Because
of this, GABA genes are of interest in the search for causes
of drug use disorders. A dinucleotide repeat (DNR)
polymorphism of the GABA receptor β3 subunit gene
(GABRB3) results in either the presence of the 181-bp G1
or 11 other repeats designated as non-G1 (NG1). Research
indicates that the NG1 is more prevalent in children of
alcoholics (COAs; Namkoong, Cheon, Kim, Jun, & Lee,
2008). Presence of the NG1 has been associated with
alcohol dependence (Noble et al., 1998; Song et al., 2003).
In addition, other GABA receptor genes have also been
associated with this disorder (Edenberg et al., 2004).
To date, the replication of research that indicates that
single polymorphisms predict drug use has been inconsis-
tent, leading to confusion and, at times, lack of confidence in
findings of genetic vulnerability to drug use disorders. This
has also been true for association of single genes with other
complex behavioral disorders (NCI-NHGRI Working Group
on Replication in Association Studies et al., 2007). Although
population stratification has been the popular scapegoat for
the mixed findings in gene association studies (Hutchison,
Stallings, McGeary, & Bryan, 2004), there are a number of
other reasons that might explain these difficulties (Colhoun,
McKeigue, & Smith, 2003; Nestler, 2000).
One reason is that any single gene is likely to have a
relatively small effect on a complex disorder, making
experimental detection difficult (Nestler, 2000). A recent
study (Berggren et al., 2006), using the largest number of
participants to date, found that the TaqI A A1+ genotype was
significantly associated with alcohol dependence; however,
the effect size was small. Mixed findings are expected when
dealing with small effect sizes. The same holds true for the
association of any single gene with other behavioral
disorders (NCI-NHGRI Working Group on Replication in
Association Studies et al., 2007). Further, complex gene–
gene and gene–environment interactions, although frequent-
ly hypothesized as the causal mechanism of drug use
disorders (Colhoun et al., 2003; NCI-NHGRI Working
Group on Replication in Association Studies et al., 2007;
Nestler, 2000), are rarely tested. However, when they are
tested, the findings support the notion that complex gene–
gene relationships may account for inconsistent findings
across different single gene studies (Yang et al., 2008).
Another reason for the difficulties in replicating single-
gene associations with drug use disorders is sex-based
differences in neurochemistry and neuroanatomy (de
Courten-Myers, 1999). More specifically, significant differ-
ences have been found in brain dopaminergic activity
between males and females (Kaasinen, Nägren, Hietala,
Farde, & Rinne, 2001; Pohjalainen, Rinne, Nägren,
Syvälahti, & Hietala, 1998). These sexually dimorphic
brain differences could explain differential prevalence of
drug use disorders between sexes (Feldstein & Miller, 2006;
Limosin et al., 2002). Studies that do not consider sex of the
participant could fail to model the complicated association of
dopaminergic genes with drug use disorders.
A number of additional variables have been reported as
significant predictors of drug use disorders. Among these
are personality traits, positive and negative life events,
negative affect, and neurocognitive functioning (Berman &
Noble, 1995; Conner et al., 2005; Wills, Vaccaro, &
McNamara, 1992). This further complicates etiological
understanding of drug use disorders because any or all of
these variables could interact with genes to better explain
the causal mechanisms underlying the development of
these disorders. Thus, rather than studying the association
of any single gene with drug use, as has been the
prevailing zeitgeist, researchers should study associations
among complex gene–gene and gene–nongene (e.g.,
psychological, neurological, and environmental variables,
which may have their own genetic basis) interactions and
drug use disorders.
The goal of this preliminary, or pilot, study was to
explore the complex relationships among genetic, personal-
ity, mood, neurocognitive, and environmental variables that
contribute to the development of drug use to test the
hypodopaminergic functioning hypothesis proposed above
in a sample of adolescent COAs. This is a good sample for
this exploratory research, as COAs are at an increased risk
for developing drug use disorders when compared to
children whose parents are not dependent on alcohol
(Heath et al., 1997). It is expected, given the previous
research described above, that predictors will vary based on
sex of the participant; so, in addition to conducting this
exploratory analysis within the total sample, similar analyses
were run on the two sexes separately.
2. Materials and methods
2.1. Participants
Participants' parents were primarily recruited through
advertisements on alcoholism treatment center bulletin
boards and in treatment community newspapers. Parents
who were interested in having their children participate
contacted the UCLA Alcohol Research Center. On this
telephone call, they were given more detailed information
and a brief initial screening interview. During a subsequent
interview conducted at the UCLA Alcohol Research
Center, the Structured Clinical Interview for DSM-IV
Disorders (First, Spitzer, Gibbon, & Williams, 1996) was
administered to all parents to establish the presence or
absence of alcohol dependence as well as the presence or
absence of other drug abuse/dependence and psychiatric
disorders. To be included in the study, participants had to
have at least one parent who met the Diagnostic and
Statistical Manual of Mental Disorders, Fourth Edition
(DSM-IV-TR; American Psychiatric Association, 2000)
criteria for alcohol dependence.
 B.T. Conner et al. / Journal of Substance Abuse Treatment 38 (2010) 178–190
As part of the overall study protocol, adolescent
participants were excluded from the study if they had
hearing or visual impairment (unless corrected to normal),
color blindness, or a history of head injury with a loss of
consciousness greater than 5 minutes or other evidence of
organic brain damage. These exclusions were necessary
because of the event-related potential electroencephalogra-
phy data that were collected but were not used in this study.
Ethnicity/Race was reported by the COAs' parents about
themselves, their children, and their parents. In this study,
to control for population stratification effects, the sample
used was restricted to non-Hispanic Caucasian participants
of European descent. This left a final sample of 111
Caucasian adolescent COAs (57 males and 54 females,
mean age = 14.5 years).
All children, as well as their parents, gave informed
assent. Participants were monetarily compensated for their
participation. The research protocol had the approval of the
UCLA Institutional Review Board.
2.2. Genotyping procedures
A 10-ml blood sample was drawn from each participant.
The amplification of DNA was carried out using a Perkin
Elmer GeneAmp 9600 thermocycler.
2.2.1. ANKK1 TaqI A SNP
The ANKK1 TaqI A polymorphism was determined
using a standard polymerase chain reaction (PCR) procedure
(Grandy, Zhang, & Civelli, 1993). The PCR product was
digested with TaqI restriction endonuclease, and the resultant
products were resolved by 2% agarose gel electrophoresis.
Two alleles were obtained: the A1 allele (the uncleaved 310-
bp fragment) and the A2 allele (the cleaved 180- and 130-bp
fragments). The presence of the A1 homozygote (A1/A1) or
heterozygote (A1/A2) was considered a genetic risk marker
for hypodopaminergic functioning compared to the A2
homozygote (A2/A2).
2.2.2. DRD2 C957T SNP
The DRD2 C957T SNP was determined by a standard
PCR procedure (Duan et al., 2003). The PCR product was
digested with TaqI restriction endonuclease, and the resultant
products were resolved by 2.5% agarose gel electrophoresis.
Two alleles were obtained: the T allele (uncleaved 196-bp
fragment) and the C allele (the cleaved 174- and 22-bp
fragments). The presence of the T homozygote (T/T) or
heterozygote (T/C) was considered a genetic risk marker for
hypodopaminergic functioning compared to the C homozy-
gote (C/C).
2.2.3. DRD4 Exon III VNTR
The DRD4 VNTRs were determined using a standard
PCR procedure (Lichter et al., 1993). The VNTRs in the
PCR product were resolved by 2.5% agarose gel electro-
phoresis. A number of VNTRs were obtained. The repeats
181
(R) included the 2R (379 bp), 3R (427 bp), 4R (475 bp), 5R
(523 bp), 6R (571 bp), 7R (619 bp), and 8R (667 bp) alleles.
The presence of the 7R homozygote (7R/7R) or heterozygote
(7R/N7R) was considered a genetic risk marker for
hypodopaminergic functioning compared to the absence of
the 7R (N7R/N7R).
2.2.4. Dopamine transporter (DAT1) VNTR
The DAT1 VNTRs were determined by a standard PCR
procedure (Vandenbergh et al., 1992). The VNTRs in the
PCR product were resolved by 2.5% agarose gel electro-
phoresis. VNTRs of interest included the 7R (360 bp), 8R
(400 bp), 9R (440 bp), 10R (480 bp), 11R (520 bp), and 12R
(560 bp). The presence of the 9R homozygote (9R/9R) or
heterozygote (9R/N9R) was considered a genetic risk marker
for hypodopaminergic functioning compared to the absence
of the 9R (N9R/N9R).
2.2.5. COMT Val108Met SNP
COMT polymorphism was determined by a standard
PCR procedure (Hoda et al., 1996). The resultant PCR
product was digested by N1aIII restriction endonuclease,
and the DNA fragments resolved by 4% agarose gel
electrophoresis. The Val-108 allele gives DNA fragments of
136 and 81 bp. The Met-108 allele gives DNA fragments of
96, 81, and 40 bp. The Val homozygote (Val/Val) or
heterozygote (Val/Met) was considered a genetic risk
marker for hypodopaminergic functioning compared to the
Met homozygote (Met/Met).
2.2.6. GABRB3 CA-dinucleotide repeat
The GABRB3 CA-dinucleotide repeat polymorphism
was determined by a modified (Noble et al., 1998) PCR
procedure (Mutirangura, Ledbetter, Kuwano, Chinault, &
Ledbetter, 1992). The PCR products were resolved using
polyacrylamide gel electrophoresis and the 33P-labeled DNA
fragments revealed by autoradiography. Twelve CA-repeat
alleles of interest were designated as follows: G1 (181 bp)
and NG1: G2 (183 bp), G3 (185 bp), G4 (187 bp), G5 (189
bp), G6 (191 bp), G7 (193 bp), G8 (195 bp), G9 (197 bp),
G10 (199 bp), G11 (201 bp), and G12 (203 bp). The
presence of either the NG1 homozygote (NG1/NG1) or
heterozygote (NG1/G1) was considered a genetic risk
marker for hypodopaminergic functioning compared to the
G1 homozygote (G1/G1).
2.2.7. Genotype combinations
In addition to examining each of the genotypes listed
above, all possible genotype combinations (pairs, triplets,
quadruplets, quintuplets, and sextuplets) were entered as
predictors. In addition, the presence of each genotype that
coded for hypodopaminergic functioning was scored as 1,
and the presence of the alternative genotype was scored as 0.
Scores were then summed across locations similar to the
method used by Harlaar et al. (2005). This resulted in a count
of the number of hypodopaminergic genotypes per
182 B.T. Conner et al. / Journal of Substance Abuse Treatment 38 (2010) 178–190
participant, hereafter referred to as the hypodopaminergic
genetic risk variable.
2.3. Predictor variables
A number of predictor variables were included in this
exploratory analysis in addition to the genetic data. These
measures were selected because previous research has
suggested that these variables are significant predictors of
the onset of drug use in adolescence. Although this list of
predictors is certainly not exhaustive, it represents a robust
collection of variables that could better account for variations
in drug use in adolescence.
2.3.1. Visuospatial ability
Visuospatial ability, assessed using a computerized
version (Berman & Noble, 1995) of the Judgment of Line
Orientation Test (JLOT; Benton, Varney, & Hamsher,
1978), was used as a measure of neurocognitive
functioning. Previous research suggests links between
neurocognitive functioning and drug use (Shoal &
Giancola, 2001).
2.3.2. Personality
Psychoticism (P), extraversion (E), and neuroticism
(N) were assessed with the Eysenck Personality Ques-
tionnaire-Junior (Eysenck & Eysenck, 1975). Novelty
Seeking (NS), Harm Avoidance (HA), and Reward
Dependence (RD) were assessed with the Junior Tem-
perament and Character Inventory (Luby, Svrakic,
McCallum, Przybeck, & Cloninger, 1999). Previous
research indicates that some of these personality variables
are predictive of drug use, including P (Conner et al.,
2005; Sáiz et al., 2003), E (Merenäkk et al., 2003), and
N (Florsheim et al., 2007). Research also supports
predictive relationships between NS (Cloninger, Sigvards-
son, & Bohman, 1988; Mâsse & Tremblay, 1997), HA
(Cloninger et al., 1988; Tcheremissine, Lane, Cherek, &
Pietras, 2003), and RD (Tcheremissine et al., 2003) and
drug use in adolescence.
2.3.3. Positive and negative life events
Positive and negative life events were measured using
the Life Stressors and Social Resources Inventory-Youth
scale (Moos & Moos, 1994). These variables were
counts of positive (range = 0–45) and negative (range =
0–78) life events across multiple domains (i.e., in
school, at home) that occurred in the 12 months prior
to the interview. Previous research suggests that both
positive (Scheier, Botvin, & Miller, 1999; Wills et al.,
1992) and negative (Taylor, 2006; Wills et al., 1992) life
events are related to drug use among adolescents.
Although stress scores were also available for use as
predictor variables, almost all of the participants had
high stress scores, which significantly restricted their
utility in these analyses.
2.3.4. Depressive symptoms
Depressive symptoms were assessed using the Beck
Depression Inventory-II (BDI-II; Beck, 1996). The BDI-II is
a 21-item state-based depressive symptoms self-report
questionnaire. Multiple research studies suggest that depres-
sive symptoms are predictive of drug use (Aseltine, Gore, &
Colten, 1998; Fleming, Mason, Mazza, Abbott, & Catalano,
2008; King, Iacono, & McGue, 2004).
2.4. Outcome variable—drug use
The self-report Health Behavior Questionnaire (Jessor,
Donovan, & Costa, 1992) was administered to the
participants. On this questionnaire, participants indicated
drugs that they have tried at least once from the following
list of drugs: alcohol, nicotine, marijuana, cocaine/crack,
heroin or other opiates, phenylcyclidine, hallucinogen,
nitrous oxide, gamma-hydroxybutyric acid, amphetamine,
and barbiturates. A count variable was computed by scoring
“I have tried…” as 1 and “I have not tried…” as 0 and
summing across all drug types. This method of measuring
drug use severity in adolescents was developed by Wills
et al. (1992) and used by Conner et al. (2005) in prior
research. The number of drugs tried has been hypothesized
to be a precursor for the later development of drug use
disorders (Robins & Przybeck, 1991). Finally, recent
research conducted to create a substance use disorder
(SUD) severity scale using item response theory found that
SUD severity significantly predicted the number of different
drugs tried by the age of 19, which the authors used as a test
of construct and concurrent validity for their SUD scale
(Kirisci et al., 2006).
2.5. Statistical analyses
Data were analyzed using recursive partitioning (RP), a
type of exploratory decision tree analysis (Zhang & Singer,
1999). RP is a non- or semiparametric data mining procedure
that builds a “tree” that identifies exhaustive orthogonal
subgroups of a population, called “leaves” or “nodes,” whose
members share common characteristics that influence the
dependent variable (Lemon, Roy, Clark, Friedmann, &
Rakowski, 2003). The data are then split into smaller and
smaller nodes using a recursive procedure until each group is
perfectly homogenous. This recursive splitting takes place
separately for each “branch” of the tree.
RP is an ideal method when searching for candidate genes
and haplotypes (Zaykin & Young, 2005) and has been used
successfully in drug abuse research (Hellemann, Conner,
Anglin, & Longshore, 2009). The strength of this method is
that it allows for analysis of highly dimensional and highly
correlated data in small samples (Breiman, Freidman,
Olshen, & Stone, 1984). It detects and describes complex
interactions between predictor variables even in extreme
cases where the number of predictors is larger than the
number of independent observations. It identifies a
 B.T. Conner et al. / Journal of Substance Abuse Treatment 38 (2010) 178–190 183

parsimonious subset of predictor variables that is sufficient
to predict the specified outcome variable.
As splitting continues until perfect homogeneity is
reached, the final RP tree is likely overfit to the data from
the particular sample under investigation. The goal,
however, is to specify a model that will be applicable
to any random sample from the population. As with most
stepwise procedures, the question arises of when to stop
fitting the model to the data. The second stage of the RP
procedure consists of using cross validation to prune the
full tree to an “appropriate size” to reach this goal. This
is ideally done using the decision rules specified by the
tree on a new sample from the population (test sample)
and evaluating its ability to accurately predict group
membership. When such a dataset is not available,
however, other methods must be employed. One com-
monly used method is to bootstrap the original sample.
Bootstrapping consists of repeated reestimation of the RP
tree using random samples (with replacement) from the
original dataset. The bootstrap cross-validation procedure
estimates the cross-validated prediction error, and related
standard error, for different numbers of splits. Appropriate
tree size is usually determined by cost, model complexity,
or cross validation. Breiman et al. (1984) suggested using
a ±1 standard error rule to choose appropriate tree size. In
other words, a good-fitting tree is one with the least
number of splits and the smallest cross-validation error,
given that the tree's cross-validation error plus its
standard error is less than 1 (per definition, the relative
error of a model with no splits).
In this study, RP was used to determine the best
predictors of drug use among 75 potential predictor
variables: age, six genotypes, 57 specific combinations of
the six genotypes, the count of hypodopaminergic geno-
types, six personality scale scores, BDI-II scores, JLOT
scores, and positive and negative life events scores. The
data were analyzed in three groups: the entire sample,
males, and females, as previous research indicates that sex
could be an important moderating variable (Flannery,
Vazsonyi, Torquati, & Fridrich, 1994; Jang, Livesley, &
Vernon, 1997; Kaasinen et al., 2001; Pohjalainen, Rinne,
Nägren, Syvälahti, et al., 1998). The cut-point estimation
was based on the Poisson distribution to account for the
effect of the right skew of count data of rare events. The
choice of the splitting rule was verified by evaluating
alternative splitting rules using the cross-validated error as
the criterion. The size of the terminal nodes was not preset.
The cross validation was based on 100 bootstrapped
samples. The rpart package version 3.1-33 (Therneau &
Atkinson, 2006), as implemented in version 2.4 of the R
software package, was used to estimate the RP trees.
3. Results
Table 1 presents descriptive data on the variables in the
study. Table 2 presents the gene frequency data. Table 3
presents the correlations between each of the predictor
variables and the outcome variable. Results from Hardy–
Weinberg Equilibrium analyses revealed that genotype
frequencies in this sample did not differ significantly from
what would be expected in the general population.
3.1. Analysis of the entire sample
The RP analysis of the entire sample of COAs (n = 111)
produced a model that had a relative error = 0.77 and a cross-
validation error = 0.91 (SE = 0.16), indicating that this model
explains approximately 23% of the variance in the entire
sample and, on average, 9% in the bootstrap cross-validation
procedure. However, given the size of the standard error, the
9% variance explained is not significantly different from
zero, indicating a model with unacceptable fit. Based on
previous research, this result was not unexpected and
supported the decision to evaluate separately the two sex-
based subsamples.

Table 1
Variables in the analysis of predictors of drug use in COAs
Variable Total (N = 111) Male (n = 57) Female (n = 54)
Age, M (SD) 14.5 (1.3) 14.5 (1.2) 14.6 (1.4)
Personality, M (SD)
Psychoticism 3.06 (2.83) 4.02 (3.09) 2.07 (2.15)
Extraversion 19.07 (3.59) 19.59 (3.13) 18.54 (3.98)
Neuroticism 9.65 (4.46) 9.57 (4.72) 9.74 (4.22)
NS 8.77 (3.13) 8.48 (3.06) 9.08 (3.20)
RD 4.86 (2.19) 4.36 (2.17) 5.40 (2.10)
HA 6.56 (3.93) 6.89 (3.97) 6.21 (3.89)
Visuospatial Ability, M (SD) 23.64 (4.34) 24.40 (4.81) 22.90 (3.73)
Depressive Symptoms, M (SD) 4.07 (6.08) 3.91 (6.45) 4.24 (5.72)
Negative Life Events, M (SD) 8.52 (5.12) 9.04 (4.81) 8.00 (5.58)
Positive Life Events, M (SD) 10.06 (3.48) 10.26 (3.51) 9.85 (3.47)
Number of drugs tried
M (SD) 1.76 (1.93) 1.79 (1.72) 1.72 (2.15)
Median 1.00 2.00 1.00
184 B.T. Conner et al. / Journal of Substance Abuse Treatment 38 (2010) 178–190

Table 2 0.18). The best predictor of drug use was the hypodopami-
Genotype frequencies nergic genetic risk variable. Males who had 4 or more
Gene Total (N = 111) Male (n = 57) Female (n = 54) hypodopaminergic genotypes tried a mean of 3.02 drugs
DRD2 TaqI A SNP compared to males who had less than four hypodopaminer-
A2/A2 67 (61) 38 (67) 29 (54) gic genotypes, who tried a mean of 1.29 drugs. These two
A1/A2 39 (35) 18 (31) 21 (39) groups were further split into seven distinctive groups. Males
A1/A1 5 (4) 1 (2) 4 (7)
who had four or more hypodopaminergic genotypes and a
DRD2 C957 SNP
C/C 29 (26) 17 (30) 12 (22) RD score equal to or greater than 5.5 tried a mean of 4.27
C/T 58 (52) 28 (49) 30 (56) drugs, the highest drug use. Males who had less than four
T/T 24 (22) 12 (21) 12 (22) hypodopaminergic genotypes, a Psychoticism score of less
DRD4 exon III VNTR than 2.5, and a RD score of less than 6 tried a mean of 0.10
N7R/N7R 73 (66) 36 (63) 37 (68)
drugs, the lowest drug use.
N7R/7R 33 (30) 18 (32) 15 (28)
7R/7R 5 (4) 3 (5) 2 (4) In addition to the groups with the highest and lowest
DAT1 VNTR drug use, the male COAs were further divided into three
N9R/N9R 59 (53) 32 (56) 27 (50) intermediate nodes. Males who had less than four
N9R/9R 48 (43) 22 (39) 26 (48) hypodopaminergic genotypes and a Psychoticism Score
9R/9R 4 (4) 3 (5) 1 (2)
of 2.5 or more, and the DRD4 heterozygote (7R/N7R)
COMT Val108Met SNP
Met/Met 27 (24) 15 (27) 12 (23) tried a mean of 0.66 drugs, compared to males who had
Val/Met 62 (56) 31 (54) 31 (57) the DRD4 homozygotes (7R/7R, N7R/N7R), who had
Val/Val 22 (20) 11 (19) 11 (20) tried a mean of 2.39 drugs. The latter group was further
GABRB3 DNR a divided on their number of hypodopaminergic genotypes.
G1/G1 14 (14) 4 (8) 10 (20)
Participants who had 3 hypodopaminergic genotypes tried
G1/NG1 41 (41) 21 (42) 20 (41)
NG1/NG1 44 (45) 25 (50) 19 (39) a mean of 3.25 drugs compared to those who had less than
Hypodopaminergic genotype count three hypodopaminergic genotypes, who tried a mean of
M (SD) 3.23 (0.97) 3.14 (1.00) 3.31 (0.95) 1.07 drugs.
Median 3.00 3.00 3.00
Values are expressed as number (percentage). 3.3. Analysis of the female subsample
a
Twelve participants (seven males, five females) were missing
GABRB3 data. The RP tree for the female subsample of COAs is
presented in Fig. 2. The overall fit of the model was good in
3.2. Analysis of the male subsample that this model explained 53% of the variability in this
sample, and on average, 20% in the bootstrap cross-
The RP tree for the male subsample of COAs is shown in validation procedure (relative error = 0.47, cross-validation
Fig. 1. The overall fit of the model was good in that it error = 0.80, SE = 0.15). The best predictor of drug use was
explained 68% of the variability in this sample and, on Negative Life Events. Female COAs who reported less than
average, 21% in the bootstrap cross-validation procedure 7.5 Negative Life Events in the previous 12 months tried a
(relative error = 0.32, cross-validation error = 0.79, SE = mean of 0.85 drugs. Those that reported 7.5 or more

Table 3
Correlations among the predictors and the outcome variable
Variable combination Total (N = 111) Male (n = 57) Female (n = 54)
Age M (SD) and NDT 0.34 ⁎⁎⁎ 0.20 0.43 ⁎⁎⁎
Personality
Psychoticism and NDT 0.16 0.18 0.23
Extraversion and NDT 0.21 ⁎ 0.24 0.21
Neuroticism and NDT 0.12 0.02 0.21
NS and NDT 0.22 ⁎ 0.09 0.31 ⁎
RD and NDT 0.16 0.24 0.10
HA and NDT −0.18 −0.16 −0.19
Visuospatial Ability and NDT −0.19 −0.11 −0.25
Depressive Symptoms and NDT 0.04 −0.01 0.08
Negative Life Events and NDT 0.09 0.29 −0.08
Positive Life Events and NDT −0.19 −0.05 −0.30
Hypodopaminergic Genotype Count and NDT 0.94 0.41 ⁎⁎ −0.14
Numbers are Pearson r values. NDT = number of drugs tried.
⁎ p b .05.
⁎⁎ p b .01.
⁎⁎⁎ p b .001.
 B.T. Conner et al. / Journal of Substance Abuse Treatment 38 (2010) 178–190 185

Fig. 1. Decision tree for the male subsample (n = 57). The first number within each node represents the mean number of drugs tried for the group, and the second
number represents the size of the group. Circles represent internal nodes that are split further, and squares represent terminal nodes. Relative error = 0.32, cross-
validation error = 0.79 (SE = 0.18).

Negative Life Events in the previous 12 months tried a mean
of 2.81 drugs. The female COAs were further divided into
four separate nodes based on two additional splits. Among
female COAs that reported 7.5 or more Negative Life
Events, those who scored 3.5 or more on the HA scale tried a
mean of 1.78 drugs compared to those who scored less than
3.5 on the HA scale, who tried a mean of 5.62 drugs, the
highest drug use. Among those females that had less than 7.5
Negative Life Events, those who scored 9.5 or more on the
NS scale tried a mean of 1.62 drugs compared to those that
scored lower than 9.5 on the NS scale, who tried a mean of
0.28 drugs, the lowest drug use.

Fig. 2. Decision tree for the female subsample (n = 54). The first number within each node represents the mean number of drugs tried for the group, and the
second number represents the size of the group. Circles represent internal nodes that are split further, and squares represent terminal nodes. Relative error = 0.47,
cross-validation error = 0.80 (SE = 0.15).
186 B.T. Conner et al. / Journal of Substance Abuse Treatment 38 (2010) 178–190
4. Discussion
Using RP to search through a large set of genetic,
personality, neurocognitive, affective, and life event vari-
ables, this exploratory study was not able to identify a stable
model of predictors of drug use, nor support the hypodopa-
minergic hypothesis proposed in the introduction in the total
sample of adolescent Caucasian COAs. However, stable
models were identified for males and females when analyzed
separately. In males, hypodopaminergic genetic risk was the
salient predictor of the number of drugs tried, whereas in
females, it was an environmental factor, Negative Life
Events. These findings are consistent with a behavioral–
genetic study (Jang et al., 1997) of twins that showed that
drug use in males was significantly determined by genetic
factors, whereas in females, it was wholly determined by
environmental factors. This finding is also consistent with
additional research suggesting significant differences in the
pathways to drug use between males and females (Derringer,
Krueger, McGue, & Iacono, 2008; Han et al., 1999;
Kashdan, Vetter, & Collins, 2005).
In this study, the decision tree for males identified the
number of hypodopaminergic genotypes as the most
important variable in determining drug use. The other
important variables in the male model were the personality
traits Psychoticism and Reward Dependence, with higher
scores on each being predictive of a greater number of drugs
tried. Individuals who score high compared to those who
score average to low on measures of Psychoticism tend to
have less control over their emotions and exhibit excessive
aggressive or impulsive behaviors (Eysenck & Eysenck,
1975). Psychoticism has been shown to be positively
correlated with drug use and drug use disorders (Conner
et al., 2005). Individuals who score high compared to those
who score low on measures of Reward Dependence tend to
show a greater bias in the maintenance of behavior in
response to cues of social reward (Cloninger, 1987).
Previous research has found a positive predictive relation-
ship between higher reward dependence and drug use among
adolescents (Deas & Thomas, 2002).
For the females, two personality traits, NS and HA, were
identified as important in predicting drug use after dividing
the sample based on the number of Negative Life Events they
had experienced. These splits were not surprising, as
previous studies have found higher NS and lower HA in
children to be strongly predictive of drug use and abuse
(Cloninger et al., 1988).
The present exploratory study supported the hypothesis
developed in the introduction by identifying hypodopami-
nergic genotypes as the best predictor of drug use behavior in
male adolescent COAs. It is in accord with prior research that
identified a hypodopaminergic state as a causal mechanism
in the development of drug use disorders (Melis et al., 2005).
It is also consistent with recent functional Magnetic
Resonance Imaging (fMRI) studies indicating that lower
DRD2 levels, determined by hypodopaminergic A1 geno-
type, affected performance on reward-based learning tasks
(Cohen, Krohn-Grimberghe, Elger, & Weber, 2007) and
individual ability to learn from errors (Klein et al., 2007).
This indicates that decreased sensitivity to negative con-
sequences may help explain increased risk for drug use
disorders. The findings of this study are also bolstered by the
research that indicates that hyperdopaminergic functioning
serves as a protective factor against the development of drug
use disorders (Volkow et al., 2006).
The limitations of this study should be considered when
interpreting the findings. This study was exploratory,
meaning that replication of the findings in confirmatory
studies needs to occur prior to extensive inference of the
meaning of these relationships. The sample was composed of
Caucasian adolescent COAs, limiting the generalizability of
the findings to this population. In addition, there is the
possibility that certain nonmeasured or nonincluded vari-
ables could explain the findings reported herein, as with all
nonexperimental studies. An additional limitation of this
study was the criterion variable. Although previous research
suggests that the number of drugs tried is a good proxy
measure of substance use severity (research reviewed
above), quantity or frequency of drug use may have provided
more relevant information on adolescent substance use
severity. Unfortunately, because this sample was composed
of young adolescents, the frequency and quantity of use
variables were too highly skewed and poorly distributed to
be of use in these analyses. Future research should include
these variables as outcomes whenever possible. Using an
SUD scale (e.g., Kirisci et al., 2006) may also improve the
generalizability of these findings.
Additional ways to build on the current results in future
research would be to include other candidate genes that
influence how reward is interpreted in the brain, additional
measures of environmental factors that affect drug use
behaviors, and more comprehensive measures of neurocog-
nitive functioning in a confirmatory analysis. Future
analyses should also specifically test the hypodopaminergic
hypothesis in multiple and varied samples to continue to
provide support for hypodopaminergic functioning as a
causal mechanism for the onset of drug use and the
development of drug use disorders.
It is also important to consider the strengths of this study.
One of the strengths is the use of RP in the analysis of the
data. This allowed for the identification of a pattern of
relationships that predicts drug use, which not only holds
true for this modest sample but generalizes to the entire
population from which this sample was drawn, as shown by
the boot-strapped cross validation.
An additional strength of this study is that this is the first
time that the cumulative effect of multiple genotypes coding
for hypodopaminergic functioning, regardless of their
genomic location, has been identified as an important
predictor of drug use in males. This has important implica-
tions regarding the mixed findings observed when only single
genes are studied. The findings suggest that a source of
 B.T. Conner et al. / Journal of Substance Abuse Treatment 38 (2010) 178–190
variation in the predictive relationship from any single
candidate gene to drug use, beyond itself, is the variation of
other hypodopaminergic genotypes. In other words, in any
study assessing a single gene, the strength of the predictive
relationship or its effect size, will depend on the distribution
of the unobserved hypodopaminergic genotypes. Variations
in these distributions across different studies are a likely
cause of nonreplicable single gene findings. Thus, studying
multiple polymorphic locations that lead to hypodopaminer-
gic functioning in the reward pathway should lead to larger
effect sizes and increased replicability of findings of the
genetic contribution to drug use disorders. In addition, at least
for males, this study supports a hypothesis that describes the
mechanism by which genetic polymorphisms contribute to
drug seeking and drug use, not just associations between
genotypes and complex human behavior.
A further strength of this study was the identification of
sex differences and the development of two separate
predictive models. Males with hypodopaminergic function-
ing are more likely to use drugs that stimulate the
mesocorticolimbic system than those with normal dopami-
nergic functioning. In contrast, among females, those living
in environments rife with negative events are at increased
risk for using more drugs and more types of drugs, which
increases their risk of developing drug use disorders. These
findings also have important theoretical implications. First,
males and females may start using drugs and develop drug
use disorders for entirely different reasons. Second, studies
that fail to consider these sex differences may have difficulty
detecting significant findings or may misrepresent the
etiology of drug use disorders if the sample is dominated
by one sex.
There are practical implications of both the method used
and the findings of this study. Identifying those at risk prior
to the onset of drug use disorders allows for early diversion
and prevention and, thus, avoidance of the negative
outcomes associated with such use. The decision trees
generated by RP identify those at increased risk for
developing drug use disorders during an early stage of
their lives. In addition, the results suggest that different
groups of at-risk individuals would benefit from interven-
tions that are specifically tailored to their risk profile.
Acknowledgments
This research was supported by the Christopher D.
Smithers Foundation, Inc., New York. The authors would
like to thank Ms. Bonita Porch for her editorial assistance.
This study was presented in part at the 115th Annual
Convention of the American Psychological Association, San
Francisco, CA, August 19, 2007.
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