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Association, interaction, and replication analysis of genes encoding serotonin transporter and 5-HT3 receptor subunits A and B in alcohol dependence

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Abstract

On the basis of the converging evidence showing regulation of drinking behavior by 5-HT3AB receptors and the serotonin transporter, we hypothesized that the interactive effects of genetic variations in the genes HTR3A, HTR3B, and SLC6A4 confer greater susceptibility to alcohol dependence (AD) than do their effects individually. We examined the associations of AD with 22 SNPs across HTR3A, HTR3B, and two functional variants in SLC6A4 in 500 AD and 280 healthy control individuals of European descent. We found that the alleles of the low-frequency SNPs rs33940208:T in HTR3A and rs2276305:A in HTR3B were inversely and nominally significantly associated with AD with odds ratio (OR) and 95 % confidence interval of 0.212 and 0.073, 0.616 (P = 0.004) and 0.261 and 0.088, 0.777 (P = 0.016), respectively. Further, our gene-by-gene interaction analysis revealed that two four-variant models that differed by only one SNP carried a risk for AD (empirical P < 1 × 10−6 for prediction accuracy of the two models based on 106 permutations). Subsequent analysis of these two interaction models revealed an OR of 2.71 and 2.80, respectively, for AD (P < 0.001) in carriers of genotype combinations 5′-HTTLPR:LL/LS(SLC6A4)–rs1042173:TT/TG(SLC6A4)–rs1176744:AC(HTR3B)–rs3782025:AG(HTR3B) and 5′-HTTLPR:LL/LS(SLC6A4)–rs10160548:GT/TT(HTR3A)–rs1176744:AC(HTR3B)–rs3782025:AG(HTR3B). Combining all five genotypes resulted in an OR of 3.095 (P = 2.0 × 10−4) for AD. Inspired by these findings, we conducted the analysis in an independent sample, OZ-ALC-GWAS (N = 6699), obtained from the NIH dbGAP database, which confirmed the findings, not only for all three risk genotype combinations (Z = 4.384, P = 1.0 × 10−5; Z = 3.155, P = 1.6 × 10−3; and Z = 3.389, P = 7.0 × 10−4, respectively), but also protective effects for rs33940208:T (χ 2 = 3.316, P = 0.0686) and rs2276305:A (χ 2 = 7.224, P = 0.007). These findings reveal significant interactive effects among variants in SLC6A4–HTR3A–HTR3B affecting AD. Further studies are needed to confirm these findings and characterize the molecular mechanisms underlying these effects.

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Acknowledgments

This study was funded by grants from the National Institute on Drug Abuse to MDL (Grants R01 DA012844 and 5 R01DA013783) and from the National Institute on Alcohol Abuse and Alcoholism to BAJ (Grants 7 U10 AA011776-10, 1 N01 AA001016-000, 7 R01 AA010522-12, and 5 R01 AA012964-06) and NA-D (Grant 5 K23 AA000329-06). We are grateful to Dr. Shaolin Wang for his help with the statistical analyses. Additionally, we are thankful to the NIH GWAS data repository for providing us access to their dataset, through project 771 to Ming D. Li, under the title of “Genome-wide association analysis for addiction and type 2 diabetes”. GWAS of Alcohol Research using Australian twins and their families (OZ-ALC): Funding support for the [CIDR-OZ-ALC GWAS] was provided through the [Center for Inherited Disease Research (CIDR)] and the National Institute on Alcohol Abuse and Alcoholism (NIAAA). [CIDR-OZ-ALC GWAS] is a genome-wide association study funded as part of NIAAA grant 5 R01 AA013320-04. Assistance with phenotype harmonization and genotype cleaning, as well as with general study coordination, was provided by the [CIDR-OZ-ALC GWAS]. Assistance with data cleaning was provided by the National Center for Biotechnology Information. Support for collection of datasets and samples was provided by the MARC: Risk Mechanisms in Alcoholism and Comorbidity (MARC; P60 AA011998-11). Funding support for genotyping, which was performed at the Johns Hopkins University Center for Inherited Disease Research, was provided by the NIH GEI (U01HG004438), the NIAAA, and the NIH contract “High throughput genotyping for studying the genetic contributions to human disease” (HHSN268200782096C).

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Authors and Affiliations

  1. Department of Psychiatry and Neurobehavioral Sciences, University of Virginia, 1670 Discovery Drive, Charlottesville, VA, 22911, USA

    Chamindi Seneviratne, Jason Franklin, Katherine Beckett, Nassima Ait-Daoud, Bankole A. Johnson & Ming D. Li

  2. Department of Public Health Sciences, University of Virginia, Charlottesville, VA, USA

    Jennie Z. Ma

  3. ACT Center for Tobacco Treatment, Education and Research, Department of Otolaryngology, University of Mississippi Medical Center, Jackson, USA

    Thomas J. Payne

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  1. Chamindi Seneviratne
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  2. Jason Franklin
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  3. Katherine Beckett
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  5. Nassima Ait-Daoud
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Correspondence to Ming D. Li.

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Seneviratne, C., Franklin, J., Beckett, K. et al. Association, interaction, and replication analysis of genes encoding serotonin transporter and 5-HT3 receptor subunits A and B in alcohol dependence. Hum Genet 132, 1165–1176 (2013). https://doi.org/10.1007/s00439-013-1319-y

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