Genetic Validation in Alcoholism

Ru-Band Lu, MDProfessor

Department of PsychiatryNational Cheng Kung University, Taiwan

Genetic Component and Alcoholism

Family studies (Merigangas 1990)

Twin studies (Pickens et al. 1991)

Adoption studies (Cadoret et al. 1985)

Genetic component in alcoholism–More than 50 % (Reich et al. 1999)

The Importance of Subtypes Alcoholism

Heterogenesis of mental illnesses

Understanding different

pathogenesis

Different treatments and prognosis

Alcoholism Subtypes

Jellineck (1960) : 2 subtypes

Morey and Skinner (1986) : 3 subtypes

Lesch (1988) : 4 subtypes

Zucker (1987) : 4 subtypes

Cloninger (1987) : 2 subtypes

Babor (1992) : 2 subtypes

Limitations

Lack of cutting point

Different risk factors in different

cultures

Inconvenience for clinical use

Lack of genetic validation

Clinical Subtyping of Alcoholism

(Huang et al., 2004)

Alcoholic patient often meeting most of the diagnostic criteria (DSM-IV-TR, 2000)

More homogeneous mental illness from the diagnostic criteria perspective.

Higher frequent comorbidity with other mental illnesses

Clinical Subtyping of Alcoholism

(Huang et al., 2004)

Pure Alcoholism

Anxiety-Depression Alcoholism

Antisocial Alcoholism

Mixed Alcoholism

Clinical Subtypes of Alcoholism

Pure Alcoholism (Pure ALC)– late onset – without comorbidity with mental illnesses– less alcohol related problems

Anxiety- Depression Alcoholism (ANX/DEP ALC)– early adult onset– comorbid anxiety or depression– heavy alcohol consumption

Clinical Subtypes of Alcoholism

Antisocial Alcoholism (Antisocial ALC) – adolescent onset (very early onset)– comorbid antisocial personality disorder– more alcohol related problems

social consequence

Mixed Alcoholism (Mixed ALC)– comorbid major mental illnesses– comorbid multiple substance disorder– Alcoholism with bipolar disorders among Han

Chinese

DRD2 Gene and Alcoholism (Blum et al. 1990)

An important candidate gene of alcoholism

Controversial results or borderline significance (Reich et al., 1999)

No association with non-subtyped alcoholism among Han Chinese population in Taiwan (Lee et al., 1999)

A: Alcoholics; C: Controls; P: Percentage; F: Allele Frequency

Author (A : C)Alcoholism, N (%) TaqI-A Non-alcoholism (n,%)

A1A1 A1A2 A2A2 A1 (P) A1 (F) A1A1 A1A2 A2A2 A1 (P) A1 (F)

Association

Blum et al. 1990 (22:24)Neiswanger et al.1995(52:30)Noble et al.2000 (92:85)Kono et al. 1997,(21: 93)Ishiguro et al.1998(209:152)

1 (4.5) 13 (59.1) 8 (36.4) 63.6 34.1 0 (0) 4 (16.7) 20

(83.3) 16.7 8.3

1 (2.0) 28 (53.8)

23 (44.2) 55.8 28.8 0 (0) 4 (13.3) 26

(86.7) 13.3 6.7

6 (6.5) 44 (47.8)

42 (45.7) 54.3 30.4 3 (3.5) 13 (15.3) 69

(81.2) 18.8 11.2

6 (28.6) 7 (33.3) 8 (38.1) 61.9 45.2 10 (10.8) 49 (52.7) 34

(36.5) 63.5 37.1

34 (16) 111 (53) 64 (31) 69 43 21 (14) 64 (42) 67 (44) 56 35

No Association

Bolos et al. 1990(40:127) Heinz et al.1996 (97:113)Anghelescu et .2001(243:98)Goldman et al.1998 (276:161)Lu et al. 1996, (20:25)Lee et al. 1999, (128:85)

2 (5.0) 13 (32.5) 25 (62.5) 37.5 21.3 8 (6.3) 30 (23.6) 89

(70.1) 29.9 18.1

3 (3.1) 31 (32) 63 (64.9) 35.1 19.8 4 (3.5) 35 (31.0) 74 (65.5) 34.5 19.0

13 (5.3) 75 (30.9) 155 (63.8) 36.2 20.8 3 (3.1) 32 (32.7) 63

(64.3) 35.8 19.4

96 (35) 130 (47) 50 (18) 82 58.3 63 (39.1) 70 (43.5) 28

(17.4) 82.6 60.3

2 (10) 10 (50) 8 (40) 60 35 3 (13) 14 (58) 7 (29 ) 71 42

28 (21.9) 26 (52) 33 (25.8) 73.9 48 11

(12.9) 44 (51.8) 30 (35.5) 64.7 38.8

DRD2 Gene and Alcoholism

(Huang et al.,2004)

Genes and Clinical Subtypes of Alcoholism

DRD2 Gene and Subtypes of Alcoholism

Association– Anxiety-Depression Alcoholism (Huang et al., 2004)

No Association – Pure Alcoholism (Huang et al., 2004)

– Antisocial Alcoholism and Non-Alcoholism (Lu et al., 2002)

– Mixed Alcoholism (data not shown)

DRD2 TaqI A and TaqI B Haplotypes Chinese Han Alcohol Dependence in ANX/DEP, and Controls

GroupHaplotype Number

(2n)

Haplotype frequency(%)p-valuea

Linkage disequilibrium

p-valueb

A1/B1 A1/B2 A2/B1 A2/B2 D D′

Controls 304 0.338 0.014 0.033 0.615 0.044c 0.2211 1.000 <0.001

Pure ALCd 142 0.387 0.000 0.014 0.599 0.366e 0.2318 0.999 <0.001

ANX/DEP ALCd 226 0.473 0.000 0.031 0.496 0.005f 0.2401 1.000 <0.001

ANX/DEPd 258 0.353 0.004 0.027 0.616 0.711g 0.2211 1.000 <0.001a p value of Fisher’s Exact testb p value of linkage disequilibrium in each of the four study groupsc Controls vs. Pure ALC vs. ANX/DEP ALC χ2 value=15.788,df=9d ANX/DEP ALC, anxiety-depressive alcohol dependence ; Pure ALC, pure alcohol dependence; ANX/DEP, anxiety-depressive disorders; p value of the haplotype difference between ANX/DEP (p=0.023, χ2 =8.324,df=3); ANX/DEP ALC vs. pure ALC (p=0.122, χ2 =4.057,df=2)e Controls vs. Pure ALC ; χ2 value=3.155,df=3f Controls vs. ANX/DEP ALC ; χ2 value=11.834 , df=3g Controls vs. ANX/DEP ; χ2 value=1.444 , df=3

(Huang et al., 2004)

Haplotype Frequency of the DRD2 TaqI A and TaqI B polymorphisms

Group

Haplotype

Number (2n)

Haplotype frequency (%)

P Value†

Linkage disequilibrium

P Value‡

A1/B1 A1/B2 A2/B1 A2/B2 D D’

Normal Controls 230 0.32 0.01 0.05 0.62 0.10a 0.20 0.94 <0.0001

Antisocial ALC§ 144 0.45 0.01 0.04 0.50 0.12b 0.23 0.97 <0.0001

Antisocial Non-ALC§ 316 0.42 0.01 0.05 0.53 0.14c 0.21 0.95 <0.0001† p value of Fisher’s Exact test‡ p value of linkage disequilibrium in each of two groupsa Antisocial ALC vs. Antisocial Non-ALC vs. Normal Controls, χ2 value= 10.65, df = 6b Antisocial ALC vs. Normal Controls, χ2 value = 5.74 df = 3c Antisocial Non-ALC vs. Normal Controls, χ2 value = 5.54 df = 3§ Antisocial ALC, subjects with antisocial personality disorder and alcohol dependence; Antisocial Non-ALC, subjects with antisocial personality disorder but without alcohol dependence

(Lu et al., 2002)

MAOA Gene and Subtype Alcoholism

Association– Pure Alcoholism (Huang et al., 2007)

– Mixed Alcoholism (Hu et al., 2013)

No Association – Anxiety-Depression Alcoholism (Huang et al.,

2007)– Antisocial ALC or Non-Alcoholism (Lu et al., 2003; Wang et al., 2007)

Frequencies of MAOA polymorphisms in Han Chinese men with alcoholism

and control subjects

Groups/genotypes Sample

size

Promoter-uNTR;no (%)

χ2 P valueSample

size

EcoRV; no.(%)

χ2 P value

3-repeat 4-repeat + -

Control subjects 197 123 (62.4) 74 (37.6) 5.160 0.160 201 165 (62.2) 76 (37.8) 5.174 0.159

Pure ALC 107 53 (49.5) 54 (50.5) 4.736 0.030 108 54 (50.0) 54 (50.8) 4.283 0.039

ANX/DEP ALC 118 72 (61.0) 46 (39.0) 0.063 0.802 118 45 (38.1) 45 (38.1) 0.003 0.954

MAO A= monoamine oxidase-A; VNTR= variable number of tandem repeats; EcoRV=restriction enzyme rs 1137070; pure ALC= pure alcohol dependence; ANX/DEP ALC= alcohol dependence and anxiety or depression or both.*Control subjects v.s. pure ALC v.s. ANX/DEP ALC; df=3+Control subjects v.s. pure ALC Control subjects v.s. ANX/DEP ALC (Huang et al., 2007)

MAOA-uVNTR and DRD2 gene interaction for the risk of BP with ALC

 Model ALC+BPB OR 95% CI p-Value

MAOA-uVNTR 3-repeat −0.839 0.432 0.198–0.942 0.035*

DRD2 TaqI A1/A1 −0.123 0.884 0.323–2.419 0.810

DRD2 TaqI A1/A2 −1.182 0.833 0.392–1.770 0.635 MAOA-uVNTR 3-repeat∗DRD2 TaqI A1/A1 1.186 3.273 0.810–13.224 0.096

MAOA-uVNTR 3-repeat∗DRD2 TaqI A1/A2

1.239 3.451 1.240–9.608 0.018**

ALC+BP: alcoholism with bipolar disorder. B, coefficients; CI, confidence interval; OR, odds ratio.Reference groups are: MAOA-uVNTR 4-repeat, DRD2 TaqI A2/A2 genotypes, and controls;covarying for age. ** pb0.05. (Hu et al., 2013)

MAOA Haplotypes for the uVNTR and EcoRV RFLP Polymorphisms in Different Groups

Sample(N)

Haplotype Number (Frequency x100%)

χ2 p-value(2R +) (3R -) (3R +) (4R -) (4R +)

Community control 77 1 (1.3) 2 (2.6) 40 (51.9) 34 (44.2) 0 (0.0) 11.58 0.480

Jail control (ASB) 38 1 (2.6) 1 (2.6) 20 (52.6) 14 (36.8) 2 (5.3)

Antisocial Non- ALC 50 1 (2.0) 1 (2.0) 27 (54.0) 19 (38.0) 2 (4.0)

Antisocial ALC 41 0 (0.0) 1 (2.4) 26 (63.4) 11 (26.8) 3 (7.3)

ASPD: Antisocial Personality Disorder without meeting the diagnosis of Alcohol DependenceAntisocial ALC: Antisocial Personality Disorder with meeting the diagnosis of Alcohol

Dependence ASB: Antisocial Behavior without meeting the diagnosis of ASPD-S-ALC or ASPD-C-ALC MAO A-uVNTR : Variable number of tandem repeat locating at upstream of the MAO A gene2R, 3R and 4R= 2 repeats, 3 repeats and 4 repeats (+) and (-) = Presence and absence of restriction sites for EcoRV (Wang et al., 2007)

ADH1B , ALDH2 Gene and Subtypes of Alcoholism

ADH1B*2 and/or ALDH2*2 Allele

Protection Pure, Anxiety-Depression, Mixed (data not shown)

and Antisocial NON-Alcoholism (Lu et al., 2005; 2012)

No protection Antisocial Alcoholism (Lu et al., 2005; 2012)

ADH1B for the Risk of Antisocial Alcoholics

Community controls vs. antisocial alcoholics

Variable Regression coefficient

Standard error Odds ratio 95%

confidence interval P-value

ADH1Ba

ADH1B*2/*2 -0.379 0.315 0.684 0.369-1.269 0.229

Age 0.082 0.021 1.086 1.043-1.131 <0.001

aCoding of ADH1B genotypes are *1/*1 and *1/ *2=0, *2/ *2=1;reference group is ADH1B *1/ *1 and *1/ *2.

(Lu et al., 2005;2012)

Dependent variable in regression model

Total ASPD (N= 297)a

(Antisocial ALC and Antisocial Non-ALC)

Antisocial ALC (N=133)b Antisocial Non-ALC (N=164)c

B Sig. Odds

ratio

95%CI B Sig. Odds

ratio

95%CI B Sig. Odds

ratio

95%CI

Lower

Bound

Upper Bound

Lower Bound

Upper

Bound

Lower Bound

Upper Bound

DRD2 A1 -.10

.78 .91 .45 1.81 .36 .45 1.43 .57 3.62

-.35 .38 .70 .32 1.55

DRD2 A2 .24 .38 1.27 .75 2.13 .43 .26 1.53 .73 3.22

.15 .60 1.16 .66 2.05

ALDH2 *1*1

-.24

.40 .79 .45 1.38 .64 .09 1.89 .90 3.96

-1.0

1

.01 .36 .18 .74

DRD2*A1+

X ALDH2*1*1

1.10

.04 3.01 1.05

8.62 .42 .52 1.53 .42 5.52

1.69

.01 5.39 1.53 18.98

DRD2*A2 X ALDH2*1*1

.10 .81 1.10 .51 2.36 -.37 .46 .69 .26 1.85

.57 .24 1.76 .69 4.50

Logistic regression of ALDH2 and DRD2 genes for ASPD, antisocial ALC and non ALC

a Antisocial Participants (Antisocial ALC and Antisocial Non-ALC) vs. Healthy Controls.b Antisocial ALC vs. Healthy Controls. c Antisocial Non-ALC vs. Healthy Controls; A1+ (*A1/*A1 or*A1/*A2) Reference category: normal control, DRD2 A2A2 genotype, ALDH2*1*2+*2*2 genotype.

(Lu et al., 2005;2012)

Specific Meanings

A candidate gene reported for many times, still controversial; this gene still the pathogenesis of the subtype mental illness

The commonly reason (Parsian et al., 1991)

phenotypic heterogeneity different definitions of control groups different racially or ethnically mixed study

populations

DRD2 gene a pathogenesis of ANX/DEP alcoholism

Specific Meanings

The ALDH2*1 risk drinking and un-subtype alcoholism (Chen et al. 1999)

50% of Han Chinese belonging ALDH2*1/*1 but nearly

100% in Western populations (Agarwal and

Goedde, 1992)

ASPD only 0.1 - 0.3% in Han Chinese (Hwu et al. 1988), but

3.3% among Caucasian Americans (Goodwin &

Hamilton 2003)

Specific Meanings

70% ASPD with alcoholism higher than 15-20% alcoholism from community in Western population (Goodwin & Hamilton 2003)

6% ASPD with alcoholism (from jail) higher than1.5-3%alcoholism from community in Han Chinese (Hwu et al. 1988; Lu et al., 2013)

ASPD demonstration higher prevalence rate of alcoholism than general population in Han Chinese and Western populations

Alcoholism one of the symptoms of ASPD?

Specific Meanings

A Dutch family with a MAOA deficiency in exon 8, male demonstration severe aggressive behavior and borderline mental retardation (Brunner et al., 1993)

Difference genotype frequencies in exons 8 and 14 of MAOA genes Higher ASPD with alcoholism than controls (Parsian., 1999)

MAOA 3-repeat & early childhood maltreatment development higher antisocial behavior in adult male (Capri., 2002)

Specific Meanings

Aggressive and/or antisocial behavior, the major

symptoms of ASPD, but not the diagnosis of ASPD

Probable reasons for the positive or negative finding

of ASPD lack of antisocial non-alcoholism as a

controls

(Lu et al., 2005;

2012)

The Roles of MAO, ADH and ALDH in Dopamine

Catabolism

Dopamine Metabolism

ARs

ADH

DOPET

DOPET: 3,4-dihydroxyphenylethanolARs : aldose (aldehyde) reductaseDOPAL:3,4-dihydroxyphenylacetaldehydeCOMT:catechol-O-methyltransferase

DOPAL

(Feldman et al., 1997)

Gene to Gene and Clinical Subtypes of

Alcoholism

ALDH2 and MAOA Gene

Association

– MAOA-uVNTR 3-repeat and ALDH2 gene

Antisocial Alcoholism

Antisocial Non-Alcoholism

– ALDH2 x MAOA in ANX/DEP, Antisocial & Mixed alcoholism (Lee et al., 2009; 2010; Hu et al., 2013)

Antisocial ALC and Non-ALC after Stratification for

MAOA-uVNTR polymorphismsGene and Group Sampl

e Size (n)

Genotype number (Frequency, %)

χ2 p-value Odds ratio

95% Confiden

ce IntervalALDH2*1/

*1ALDH2*1/*2+*2/*2

MAOA 3-repeat

Antisocial ALC 81 49(60.5) 32(39.5) 18.787

1.46E-05

3.913 2.085-7.344

Antisocial Non-ALC

96 27(28.1) 69(71.9)

MAOA 4-repeat

Antisocial ALC 51 26(51) 25(49) 1.178 0.278 1.502 0.719-3.137

Antisocial Non-ALC

66 27(40.9) 39(59.1)Breslow-Day Homogeneity test of odds ratio: χ2=3.772, df=1, p=0.052 (Lee et al., 2009)

Logistic regression of interaction for MAOA and ALDH2 gene and the

risk of ANX/DEP ALC

B SE p-value Odds Ratio

95% Confidence Interval for Odds ratio

Lower Bound

Upper Bound

MAOAa -.519 .426

. .233 .595 .258 1.372

ALDH2b -2.142 .412

2.0E-7 .117 .052 .263

ALDH2 *MAOA

1.017 .512

.036 2.918 1.070 7.962

B, coefficients; SE, standard errors; p<0.05 is considered significantReference group: normal control, MAOA-uVNTR 3 allele, ALDH2 *1*1 genotypeaMAOA-uVNTR 4-repeat to 3-repeat allelebALDH2 *1*2+*2*2 to *1*1 genotypeDRD2 x MAOA x ALDH2 gene in ANX/DEP Alcoholism (Lee et al., 2010)

Logistic regression analysis of MAOA gene and ALDH2 gene and their interaction for the risk of Antisocial ALC

a. The reference category is: Antisocial Non ALC, MAOA 4-repeat, ALDH2*1/*2+*2/*2

Antisocial ALC vs Antisocial non ALCa

B df. Sig. Exp (B)

95% Confidence Interval for Exp (B)

Lower Bound Upper Bound

ALDH2 .383 1 .313 1.466 .697 3.084

MAOA -.410 1 .227 .663 .341 1.291

ALDH2 * MAOA 1.074 1 .032 2.927 1.099 7.795

(Lee et al., 2009)

MAOA-uVNTR and DRD2 gene interaction for the risk of (bipolar disorder) BP with ALC

 Model ALC+BPB OR 95% CI p-Value

MAOA-uVNTR 3-repeat −0.839 0.432 0.198–0.942 0.035

DRD2 TaqI A1/A1 −0.123 0.884 0.323–2.419 0.810

DRD2 TaqI A1/A2 −1.182 0.833 0.392–1.770 0.635 MAOA 3-repeat∗DRD2 TaqI A1/A1 1.186 3.273 0.810–13.224 0.096

MAOA 3-repeat∗DRD2 TaqI A1/A2

1.239 3.451 1.240–9.608 0.018

ALC+BP: alcoholism with bipolar disorder. B, coefficients; CI, confidence interval; OR, odds ratio.Reference groups are: MAOA-uVNTR 4-repeat, DRD2 TaqI A2/A2 genotypes, and controls;covarying for age.. (Hu et al., 2013)

ALDH2 and DRD2 Gene Interaction

Association

– ALDH2*1/*1, DRD2 A1/A1 genotypes and ANX/DEP

Antisocial Non-Alcoholism and ASPD (Huang et al.,2004; Lu

et el.,2012)

– ALDH2 * DRD2 gene in ASPD and Antisocial Non-

Alcoholism

(Lu et el., 2012)

No Association

– ALDH2 & DRD2 gene and Pure (Huang et el., 2004)

– Mixed Alcoholism (data not shown)

VariableANX/DEP ALC Pure ALC

Odds ratio 95% CI P-value Odds

ratio 95% CI P-value

ALDH2* 1/* 1

DRD2A1/A1 3.58 1.36-9.48 0.010 1.565 0.47-5.22 0.466

DRD2A1/A2 2.10 1.02-4.32 0.043 2.278 0.99-5.23 0.052

ALDH2* 1/*2&*2/*2

DRD2A1/A1 1.60 0.43-6.04 0.485 0.857 0.15-5.01 0.864

DRD2A1/A2 2.45 0.85-7.060 0.096 1.572 0.46-5.39 0.472

Multiple Logistic Regression Analysis of the DRD2 TaqI A for Risk of Alcohol Dependence with Stratification of ALDH2 genotypes

The genotype of ALDH2* 2/* 2 was not found in the alcohol-dependence subjects.Reference group is DRD2 A2/A2.Model 1:Neither controlling for age, gender nor stratification of ADH1B and Model2:Stratification of ADH1B and controlling for gender and age. Model 3:Stratification of ALDH2 and controlling for gender and age.

(Huang et al., 2004)

Genotype Subgroup DRD2 genotype distribution (and %) Χ2 p value

A1A1 A1A2 A2A2Antisocial ALC vs. ControlsALDH2*1/*1 Antisocial ALC 16(21.0) 30(39.5) 30(39.5) 4.211 0.122

Normal control 12(10.6) 46(40.7) 55(48.7)ALDH2*1/*2+*2/*2 Antisocial ALC 12(21.0) 29(50.9) 16(28.1) 1.530 0.465

Normal control 24(18.3) 58(44.3) 49(37.4)Antisocial Non ALC vs. ControlsALDH2*1/*1 Antisocial Non

ALC13(23.2) 28(50.0) 15(26.8) 9.084 0.011

Normal control 12(10.6) 46(40.7) 55(48.7)ALDH2*1/*2+*2/*2 Antisocial Non

ALC14(13.0) 55(50.9) 39(36.1) 1.649 0.483

Normal control 24(18.3) 58(44.3) 49(37.4)Antisocial personality disorder vs. ControlsALDH2*1/*1 Antisocial PD 29(22.0) 58(43.9) 45(34.1) 8.008 0.018

Normal control 12(10.6) 46(40.7) 55(48.7)ALDH2*1/*2+*2/*2 Antisocial PD 26(15.8) 84(50.9) 55(33.3) 1.298 0.522

Normal control 24(18.3) 58(44.3) 49(37.4)

DRD2TaqI A in Han Chinese Subjects after Stratification for ALDH2 polymorphisms

(Lu et al., 2010; 2011)

Dependent variable in regression model

Total ASPD (N= 297)a

(Antisocial ALC and Antisocial Non-ALC)

Antisocial ALC (N=133)b Antisocial Non-ALC (N=164)c

B Sig. Odds

ratio

95%CI B Sig. Odds

ratio

95%CI B Sig. Odds

ratio

95%CI

Lower

Bound

Upper Bound

Lower Bound

Upper

Bound

Lower Bound

Upper

Bound

DRD2 A1 -.10 .78 .91 .45 1.81 .36 .45 1.43 .57 3.62 -.35 .38 .70 .32 1.55

DRD2 A2 .24 .38 1.27 .75 2.13 .43 .26 1.53 .73 3.22 .15 .60 1.16 .66 2.05

ALDH2 *1/*1 -.24 .40 .79 .45 1.38 .64 .09 1.89 .90 3.96 -1.01 .01 .36 .18 .74

DRD2*A1/+ A1 * ALDH2*1/*1

1.10

.04 3.01 1.05 8.62 .42 .52 1.53 .42 5.52 1.69 .01 5.39 1.53 18.98

DRD2*A2 /*A2 * ALDH2*1/*1

.10 .81 1.10 .51 2.36 -.37 .46 .69 .26 1.85 .57 .24 1.76 .69 4.50

Logistic regression of ALDH2 and DRD2 genes for ASPD, antisocial ALC and non ALC

a Antisocial Participants (Antisocial ALC and Antisocial Non-ALC) vs. Healthy Controls.b Antisocial ALC vs. Healthy Controls. c Antisocial Non-ALC vs. Healthy Controls; A1+ (*A1/*A1 or*A1/*A2) Reference category: normal control, DRD2 A2A2 genotype, ALDH2*1*2+*2*2 genotype.

(Lu et al., 2010; 2011)

Specific Meanings

Stratification of ALDH2*1/*1, DRD2 gene ASPD and ANX/DEP ALC (Lu et al., 2012).

54.3% ASPD with anxiety disorder in western population

Positive correlation with ASPD and anxiety disorder. (Goodwin & Hamilton., 2003)

Anxiety disorders and ASPD associated with higher odds of alcohol use disorder (Goodwin and Hamilton, 2003)

Specific Meanings

ALDH2 * DRD2 gene and BP (Lee et al., 2010)

70-90% BP with anxiety disorder in Western, 20-40% in Han Chinese populations (Chang et al., 2012).

50% of Han Chinese belonging ALDH2*1/*1 but nearly 100% in Western populations (Agarwal and Goedde, 1992)

ALDH2 gene relation to alcoholism, ASPD, anxiety disorder & bipolar disorder.

In Summary DRD2 Association

─ ANX/DEP Alcoholism MAOA Association

─ Pure and Mixed Alcoholism ALDH2 Association

─ Pure, ANX/DEP Alcoholism and Antisocial non-alcoholism

ADH1B and ALDH2 Association─ Pure, ANX/DEP and Mixed Alcoholism

In Summary ALDH2*1/*1, DRD2 A1 /A1

− Anxiety-Depression Alcoholism− ASPD and Antisocial Non-ALC

MAOA 3-repeat ,ALDH2 Association– Antisocial Alcoholism

MAOA X ALDH2 Association– Anxiety-Depression Alcoholism– Antisocial Alcoholism– Mixed Alcoholism

ALDH2 X DRD2 Association– ASPD and Antisocial Non-ALC

Ongoing Studies

Preliminary studies of low dose (5mg) of memantine : (6 M )

Pure and anxiety-depression alcoholism

1/3 completing discontinue drinking;

1/3 markedly reduced drinking;

1/3 poor response

Antisocial and Mixed Alcoholism (bipolar alcoholism)

Poorer response than pure and anxiety-depression

alcoholism

CommentsPossible mechanism of mematine

effect Inhibition NADPH in microglial and stimulation

neurotrophic factors (BDNF etc.) in astroglial

cells

Plasma level of memantine (5mg/day): 10-50

ng/ml (mean 16

ng/ml). No effect in NMDA receptor (required > 86.3

– 431.5 ng/ml) (Parsons et al., 1999)

Conclusion More studies about MAO, ADH1B , ALDH2 and

dopamine/5-HT related genes in different ethnics of major mental illnesses

The lower prevalence of a disease, the higher statistic meaning for a fixed number of trait loci (Rice et al., 2001)

Han Chinese populations playing a specific contribution to the molecular genetic studies of alcoholism (Lu et al., 1999)

Inflammatory press & neurodegenoration involving alcoholism & substance use disorders.

Alcoholism: An important role in substance use disorder and addiction behavior

Acknowledgements

Professor Kenneth K. Kidd, Ph. D. Professor Ting-Kai Li, Ph. D. Research Scientist Judy R. Kidd, Ph.D Professor Jean-Shih Chen, Ph. D Research Scientist Andrew J. Pakstis, Ph. D. Professor Shin-Jiun Yin, Ph. D. Professor Huei-Chen Ko, Ph. D Associate Professor San-Yuan Huang, M.D., Ph.

D. Associate Professor Yaun-Hwa Chou, M.D., Ph. D. Assistant Professor Wei-Wen Lin, M.D., Ph. D. Assistant Professor Jia-Fu Lee, M.D., Ph. D.

Acknowledgements

Assistant Professor Yi-Chyan Chen, M.D., Ph. D.

Assistant Professor Michael V. Osier, Ph. D. Assistant Professor Tso-Jen Wang, M.D., Ph.

D. Hsin-Yi, Lo M.D., Ph. D. student Cheng-Yi Hahn, M.D., M.S., Ph. D. student Sheng-Yu Lee, M.D., M.S. Ms. Ju-Ping Weng, M.S. Ms. Yi-Syuan Wu, M.S. Ms. Pei-Ling Wu, M.S. Mr. Tsun-En Lu, M.S. Ms. Yu-Shan Wang.

THANK YOU FOR YOUR ATTENTION

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Drugs or toxin (Extro-neurotoxin)

Act

ivat

ion

Genetic factors(Endo-neurotoxin)

Microglia

(Working Model)

Dopaminergic

Neuronal

Damage

NADPH oxidase (PHOX)

Activated

O2O2 • –

NADPH NADP+ + H +

Resting

gp91

p22rap1a

p40

p47rac2

p67

a MAC-1

LPS

out

in

PKC

PP

Naloxone,Dextromethorphan Gly-Gly-Phe

O2

O2 • –

Dual Functions of Superoxide Radicals

gp91

Microglia

MAC1

TLR4LBP

CD14

PHOX

ROS

H2O2

O2 • –

TNF-a IL-1

NO •

ONOO -2) Gene Expression

1) Neurotoxicity

NF-kB,AP-1

PKCLPS

LPS

DA neurons

Transfection of gp91PHOX, a membrane subunit of PHOX, increases [3H]-naloxone binding

0

100

200

300

[3H]-(-)-Nal

Bin

ding

Cap

acity

(% W

T)

WT

[3H]-(+)-Nal

*

Gp91-transfected

400COS-7 Cells (WT)

Ligand Binding Assay

StableTransfection

COS-7 gp91 (gp91)

gp91

*

Serotonin Metabolism (Svensson et al. 1999)

The metabolism pattern of serotonin (5-HT). MAO = monoamine oxidase; ADH = alcohol dehydrogenase; ALR = aldehyde/aldose reductase; ALDH = aldehyde dehydrogenase; 5-HIAL = 5-hydroxyindole-3-acetaldehyde;5-HTOL = 5-hydroxytryptophol; 5-HIAA = 5-hydroxyindole-3-acetic acid

ADH1B*2 allele: protective factor

ALDH 2 *2 allele: protective factor (Thomasson et al. 1991)

Ethanol Metabolic Genes and Alcoholism

ALDH2 deficiency, which slows the elimination of acetaldehyde (Thomasson et al. 1991)

Higher acetaldehyde levels generated by the more active ADH isozymes should deter heavy drinking (Thomasson et al. 1991)

Ethanol Metabolic Genes and Alcoholism

(Thomasson et al. 1991)

a Number of individuals in group. Note that some exons did not amplify well or gave ambiguous results; thus some individuals were excluded.b Fraction of group with each genotype; because of rounding errors, some groups’ frequencies do not sum to 1.00.c Alcoholics are significantly different from nonalcoholics (P<.002). The AD H2 genotype distribution among alcoholics did not fit the Hardy-Weinberg equilibrium; all other genotype distribution did.d Alcoholics are significantly different from nonalcoholics (P<.005).

ADH and ALDHGenotype Frequencies and Allele

FrequenciesGenotype Frequencyb Allele Frequency

Group (Na) ADH1B*1/*1 ADH1B*1/*2 ADH1B*2/*2 ADH1B*1 ADH1B*2

Nonalcoholics (47)……. 0.06 0.40 0.53 0.27 0.73

Alcoholics (49)………… 0.37c 0.31c 0.33c 0.52d 0.48d

Genotype Frequencyb Allele Frequency

ALDH2*1/*1 ALDH2*1/*2 ALDH2*2/*2 ALDH2*1 ALDH2*2

Nonalcoholics (50)……. 0.52 0.36 0.12 0.70 0.30

Alcoholics (50)………… 0.88d 0.12d 0.00d 0.94d 0.06d

Contradictory Findings of ADH

ADH1B*1 among most of world population (~90%) (Bosron et al.

1993)

ADH1B*2 among Han Chinese (~70%) (Osier et al. 2002)

ADH1B*2 among Taiwanese aborigines (70-86%) (Chen et al. 1996)

Alcohol prevalence (30-50%) among Taiwanese aborigines >15 times Han Chinese in Taiwan

No support the variance of ADH1B related to rice (Kidd et al., 2010)

Ethnic genetic heterogeneity of ADH1B*2

Figure 1. Blood ethanol concentrations after administration of a low-to-moderate dose of ethanol (0.3g/kg) to men with different ADH2 and ALDH2 allelotypes during 4-h period. All subjects were homozygous ADH3*1. The genotypic groups are denoted as ADH2*1/*1, ALDH2*1/*1 (, n=12); ADH2*1/*2, ALDH2*1/*1 (, n=10); ADH2*2/*2, ALDH2*1/*1 (, n=11); ADH2*1/*1, ALDH2*1/*2 (, n=10); ADH2*1/*2, ALDH2*1/*2 (, n=12); ADH2*2/*2, ALDH2*1/*2 (, n=11). Vertical bars (only the upper or lower portion shown) represent standard errors of the means. Statistically significant differences between the groups at corresponding times by ANOVA:ap<0.001 vs. ADH2*1/*1, ALDH2*1/*1 bp<0.01 vs. ADH2*1/*2, ALDH2*1/*1cp<0.001 vs. ADH2*2/*2, ALDH2*1/*1 dp<0.05 vs. ADH2*2/*2, ALDH2*1/*2

(Peng et al. 1999)

Figure 2. Alterations of blood ethanol in men with homozygous ALDH2*1/*1 and heterozygous ALDH2*1/*2 genotypes, both included three different ADH2 allelotypes (see Fig. 1), following a low-to-moderate dose of ethanol (0.3g/kg) during 4-h period. All subjects were homozygous ADH3*1 . The genotypic groups are denoted as ALDH2*1/*1 (, n=33) and ALDH2*1/*2 (, n=33). Vertical bars (both the upper or lower portion shown) represent standard errors of the means. Statistically significant differences between the groups at corresponding times by Student’s t-test:ap<0.05 vs. ALDH2*1/*1 bp<0.01 vs. ALDH2*1/*1 cp<0.001 vs. ALDH2*1/*1

(Peng et al. 1999)

Figure 3. Blood acetaldehyde concentrations after administration of a low-to-moderate dose of ethanol (0.3g/kg) to men with different ADH2 and ALDH2 allelotypes during 4-h period. All subjects were homozygous ADH3*1. The genotypic groups are denoted as ADH2*1/*1, ALDH2*1/*1 (, n=12); ADH2*1/*2, ALDH2*1/*1 (, n=10); ADH2*2/*2, ALDH2*1/*1 (, n=11); ADH2*1/*1, ALDH2*1/*2 (, n=10); ADH2*1/*2, ALDH2*1/*2 (, n=12); ADH2*2/*2, ALDH2*1/*2 (, n=11). Vertical bars (only the upper or lower portion shown) represent standard errors of the means. Statistically significant differences between the groups at corresponding times by ANOVA:ap<0.05 vs. ADH2*1/*1, ALDH2*1/*1 bp<0.01 vs. ADH2*1/*2, ALDH2*1/*1cp<0.05 vs. ADH2*1/*2, ALDH2*1/*1 dp<0.01 vs. ADH2*1/*2, ALDH2*1/*1ep<0.05 vs. ADH2*2/*2, ALDH2*1/*1 fp<0.01 vs. ADH2*2/*2, ALDH2*1/*1

(Peng et al. 1999)

Process of DSM Diagnosis

DSM-III : reliability of mental illnesses

DSM-III-R → DSM-IV-TR–developed the validity for diagnostic

criteria