BACKGROUND: Slow neurotransmission including DARPP-32 signalling is implicated in substance use disorders (SUDs) by experimental systems but not yet in the human aetiology. PPP1R12B, encoding another protein in the DARPP-32 family, hasn't been studied in the brain. METHODS: Brain-regional gene activity was assessed in three different animal models of SUDs for mRNA level alterations. Genetic associations were assessed by meta-analysis of pre-existing dbGaP GWAS datasets for main effects and epistasis with known genetic risks, followed by cell type-specific pathway delineation. Parkinson's disease (PD) was included as a dopamine-related disease control for SUDs. FINDINGS: In animal models of SUDs, environmentally-altered PPP1R12B expression sex-dependently involves motivation-related brain regions. In humans with polysubstance abuse, meta-analysis of pre-existing datasets revealed that PPP1R12B and PPP1R1B, although expressed in dopamine vs. dopamine-recipient neurons, exerted similar interactions with known genetic risks such as ACTR1B and DRD2 in men but with ADH1B, HGFAC and DRD3 in women. These interactions reached genome-wide significances (Pmeta<10-20) for SUDs but not for PD (disease selectivity: P = 4.8 × 10-142, OR = 6.7 for PPP1R12B; P = 8.0 × 10-8, OR = 2.1 for PPP1R1B). CADM2 was the common risk in the molecular signalling regardless of gender and cell type. INTERPRETATION: Gender-dependant slow neurotransmission may convey both genetic and environmental vulnerabilities selectively to SUDs. FUNDING: Grants from National Institute on Drug Abuse (NIDA) and National Institute on Alcohol Abuse and Alcoholism (NIAAA) of U.S.A. and National Natural Science Foundation of China (NSFC).

Department of Computer Information Systems Bentley University Waltham MA 02452 United States of America

Department of Pharmacology and Experimental Therapeutics College of Pharmacy and Pharmaceutical Sciences University of Toledo Toledo Ohio 43614 United States of America

Department of Psychiatry Institute of Psychiatric Research Indiana University School of Medicine Indianapolis Indiana 46202 United States of America

Laboratory of Psychiatric Neurogenomics McLean Hospital Belmont MA 02478 United States of America

Laboratory of Psychiatric Neurogenomics McLean Hospital Belmont MA 02478 United States of America; Department of Neurology the 2nd Affiliated Hospital of Fujian Medical University Quanzhou Fujian Province P R China

National Institute on Drug Abuse and National Institute of Alcohol Abuse and Alcoholism Bethesda Maryland 20892 United States of America

School of Life Science Beijing Institute of Technology 100081 Beijing China

School of Life Science Beijing Institute of Technology 100081 Beijing China; Laboratory of Psychiatric Neurogenomics McLean Hospital Belmont MA 02478 United States of America

Zobrazit více v PubMed

Global, regional, and national comparative risk assessment of 84 behavioural, environmental and occupational, and metabolic risks or clusters of risks for 195 countries and territories, 1990-2017: a systematic analysis for the Global Burden of Disease Study 2017. Lancet. 2018;392(10159):1923–1994. PubMed PMC

Alcohol use and burden for 195 countries and territories, 1990-2016: a systematic analysis for the Global Burden of Disease Study 2016. Lancet. 2018;392(10152):1015–1035. PubMed PMC

Manthey J., Shield K.D., Rylett M., Hasan O.S.M., Probst C., Rehm J. Global alcohol exposure between 1990 and 2017 and forecasts until 2030: a modelling study. Lancet. 2019;393(10190):2493–2502. PubMed

Cates H.M., Benca-Bachman C.E., de Guglielmo G., Schoenrock S.A., Shu C., Kallupi M. National Institute on Drug Abuse genomics consortium white paper: coordinating efforts between human and animal addiction studies. Genes Brain Behav. 2019;18(6):e12577. PubMed PMC

Greengard P. The neurobiology of slow synaptic transmission. Science. 2001;294(5544):1024–1030. PubMed

Goodman A. Neurobiology of addiction. An integrative review. Biochemical pharmacology. 2008;75(1):266–322. PubMed

Takahashi S., Ohshima T., Cho A., Sreenath T., Iadarola M.J., Pant H.C. Increased activity of cyclin-dependent kinase 5 leads to attenuation of cocaine-mediated dopamine signaling. Proc Natl Acad Sci U S A. 2005;102(5):1737–1742. PubMed PMC

Zhu H., Lee M., Guan F., Agatsuma S., Scott D., Fabrizio K. DARPP-32 phosphorylation opposes the behavioral effects of nicotine. Biol. Psychiatry. 2005;58(12):981–989. PubMed

Nuutinen S., Kiianmaa K., Panula P. DARPP-32 and Akt regulation in ethanol-preferring AA and ethanol-avoiding ANA rats. Neurosci Lett. 2011;503(1):31–36. PubMed

Liu M., Jiang Y., Wedow R., Li Y., Brazel D.M., Chen F. Association studies of up to 1.2 million individuals yield new insights into the genetic etiology of tobacco and alcohol use. Nat Genet. 2019;51(2):237–244. PubMed PMC

Sanchez-Roige S., Palmer A.A., Fontanillas P., Elson S.L., Adams M.J., Howard D.M. Genome-Wide Association Study Meta-Analysis of the Alcohol Use Disorders Identification Test (AUDIT) in Two Population-Based Cohorts. Am J Psychiatry. 2019;176(2):107–118. PubMed PMC

Sanchez-Roige S., Palmer A.A., Clarke T.K. Recent Efforts to Dissect the Genetic Basis of Alcohol Use and Abuse. Biol. Psychiatry. 2019 PubMed PMC

Zhao J., Chen C., Bell R.L., Qing H., Lin Z. Identification of HIVEP2 as a dopaminergic transcription factor related to substance use disorders in rats and humans. Transl Psychiatry. 2019;9(1):247. PubMed PMC

Sun L., Wang C., Hu Y.Q. Utilizing mutual information for detecting rare and common variants associated with a categorical trait. PeerJ. 2016;4:e2139. PubMed PMC

Kleiber M.L., Mantha K., Stringer R.L., Singh S.M. Neurodevelopmental alcohol exposure elicits long-term changes to gene expression that alter distinct molecular pathways dependent on timing of exposure. J Neurodev Disord. 2013;5(1):6. PubMed PMC

Bell R.L., Rodd Z.A., Lumeng L., Murphy J.M., McBride W.J. The alcohol-preferring P rat and animal models of excessive alcohol drinking. Addict Biol. 2006;11(3–4):270–288. PubMed

Rice J., Coutellier L., Weiner J.L., Gu C. Region-specific interneuron demyelination and heightened anxiety-like behavior induced by adolescent binge alcohol treatment. Acta Neuropathol Commun. 2019;7(1):173. PubMed PMC

Coleman L.G., Jr., Liu W., Oguz I., Styner M., Crews F.T. Adolescent binge ethanol treatment alters adult brain regional volumes, cortical extracellular matrix protein and behavioral flexibility. Pharmacol. Biochem. Behav. 2014;116:142–151. PubMed PMC

Kuhn C. Emergence of sex differences in the development of substance use and abuse during adolescence. Pharmacol. Ther. 2015;153:55–78. PubMed PMC

Kota D., Alajaji M., Bagdas D., Selley D.E., Sim-Selley L.J., Damaj M.I. Early adolescent nicotine exposure affects later-life hippocampal mu-opioid receptors activity and morphine reward but not physical dependence in male mice. Pharmacol. Biochem. Behav. 2018;173:58–64. PubMed PMC

Logsdon G.A., Vollger M.R., Eichler E.E. Long-read human genome sequencing and its applications. Nat Rev Genet. 2020 PubMed PMC

Volkow N.D. Personalizing the Treatment of Substance Use Disorders. Am J Psychiatry. 2020;177(2):113–116. PubMed

Liu K., Yu J., Zhao J., Zhou Y., Xiong N., Xu J. (AZI2)3′UTR Is a New SLC6A3 Downregulator Associated with an Epistatic Protection Against Substance Use Disorders. Mol. Neurobiol. 2018;55(7):5611–5622. PubMed PMC

Volkow N.D., Baler R.D. Addiction science: uncovering neurobiological complexity. Neuropharmacology. 2014;76 Pt B:235–249. PubMed PMC

Volkow N.D., Wise R.A., Baler R. The dopamine motive system: implications for drug and food addiction. Nature reviews Neuroscience. 2017;18(12):741–752. PubMed

Paxinos G F.K. Elsevier Academic Press; San Diego, CA: 2001. The mouse brain in stereotaxic coordinates: second edition.

Salay L.D., Ishiko N., Huberman A.D. A midline thalamic circuit determines reactions to visual threat. Nature. 2018;557(7704):183–189. PubMed PMC

Jasinska A.J., Chen B.T., Bonci A., Stein E.A. Dorsal medial prefrontal cortex (MPFC) circuitry in rodent models of cocaine use: implications for drug addiction therapies. Addict Biol. 2015;20(2):215–226. PubMed PMC

Meye F.J., Soiza-Reilly M., Smit T., Diana M.A., Schwarz M.K., Mameli M. Shifted pallidal co-release of GABA and glutamate in habenula drives cocaine withdrawal and relapse. Nat. Neurosci. 2016;19(8):1019–1024. PubMed

Kim H.J., Lee J.H., Yun K., Kim J.H. Alterations in Striatal Circuits Underlying Addiction-Like Behaviors. Mol Cells. 2017;40(6):379–385. PubMed PMC

Charan J., Kantharia N.D. How to calculate sample size in animal studies? J Pharmacol Pharmacother. 2013;4(4):303–306. PubMed PMC

Rifas L., Towler D.A., Avioli L.V. Gestational exposure to ethanol suppresses msx2 expression in developing mouse embryos. Proc Natl Acad Sci U S A. 1997;94(14):7549–7554. PubMed PMC

Xu W., Huo L., Li J., Xu C., Wang S., Yang Y. Effects of Alcohol on Mitochondrial Functions of Cumulus Cells in Mice. Cell Reprogram. 2017;19(2):123–131. PubMed

Rajikin M.H., Latif E.S., Mar M.R., Mat Top A.G., Mokhtar N.M. Deleterious effects of nicotine on the ultrastructure of oocytes: role of gamma-tocotrienol. Medical science monitor: international medical journal of experimental and clinical research. 2009;15(12) BR378-83. PubMed

Asadi E., Jahanshahi M., Golalipour M.J. Effect of vitamin e on oocytes apoptosis in nicotine-treated mice. Iran J Basic Med Sci. 2012;15(3):880–884. PubMed PMC

Pan Y., Luo X., Liu X., Wu L.Y., Zhang Q., Wang L. Genome-wide association studies of maximum number of drinks. J Psychiatr Res. 2013;47(11):1717–1724. PubMed PMC

Bierut L.J., Agrawal A., Bucholz K.K., Doheny K.F., Laurie C., Pugh E. A genome-wide association study of alcohol dependence. Proc Natl Acad Sci U S A. 2010;107(11):5082–5087. PubMed PMC

Liu Q.R., Canseco-Alba A., Zhang H.Y., Tagliaferro P., Chung M., Dennis E. Cannabinoid type 2 receptors in dopamine neurons inhibits psychomotor behaviors, alters anxiety, depression and alcohol preference. Sci Rep. 2017;7(1):17410. PubMed PMC

Anderson C.A., Pettersson F.H., Clarke G.M., Cardon L.R., Morris A.P., Zondervan K.T. Data quality control in genetic case-control association studies. Nat Protoc. 2010;5(9):1564–1573. PubMed PMC

Kennedy J.L., Xiong N., Yu J., Zai C.C., Pouget J.G., Li J. Increased Nigral SLC6A3 Activity in Schizophrenia Patients: findings From the Toronto-McLean Cohorts. Schizophr Bull. 2016;42(3):772–781. PubMed PMC

Purcell S., Neale B., Todd-Brown K., Thomas L., Ferreira M.A., Bender D. PLINK: a tool set for whole-genome association and population-based linkage analyses. Am J Hum Genet. 2007;81(3):559–575. PubMed PMC

Ueki M., Cordell H.J. Improved statistics for genome-wide interaction analysis. PLoS Genet. 2012;8(4) PubMed PMC

Gu Z., Gu L., Eils R., Schlesner M., Brors B. circlize Implements and enhances circular visualization in R. Bioinformatics. 2014;30(19):2811–2812. PubMed

Hook P.W., McClymont S.A., Cannon G.H., Law W.D., Morton A.J., Goff L.A. Single-Cell RNA-Seq of Mouse Dopaminergic Neurons Informs Candidate Gene Selection for Sporadic Parkinson Disease. Am J Hum Genet. 2018;102(3):427–446. PubMed PMC

Ho H., Both M., Siniard A., Sharma S., Notwell J.H., Wallace M. A Guide to Single-Cell Transcriptomics in Adult Rodent Brain: the Medium Spiny Neuron Transcriptome Revisited. Front Cell Neurosci. 2018;12:159. PubMed PMC

Darmanis S., Sloan S.A., Zhang Y., Enge M., Caneda C., Shuer L.M. A survey of human brain transcriptome diversity at the single cell level. Proc Natl Acad Sci U S A. 2015;112(23):7285–7290. PubMed PMC

Heiman M., Schaefer A., Gong S., Peterson J.D., Day M., Ramsey K.E. A translational profiling approach for the molecular characterization of CNS cell types. Cell. 2008;135(4):738–748. PubMed PMC

Li J., Long X., Hu J., Bi J., Zhou T., Guo X. Multiple pathways for natural product treatment of Parkinson's disease: a mini review. Phytomedicine: international journal of phytotherapy and phytopharmacology. 2019;60 PubMed

Zhao Y., Xiong N., Liu Y., Zhou Y., Li N., Qing H. Human dopamine transporter gene: differential regulation of 18-kb haplotypes. Pharmacogenomics. 2013;14(12):1481–1494. PubMed PMC

Lin Z., Zhao Y., Chung C.Y., Zhou Y., Xiong N., Glatt C.E. High regulatability favors genetic selection in SLC18A2, a vesicular monoamine transporter essential for life. Faseb j. 2010;24(7):2191–2200. PubMed PMC

Lontay B., Serfozo Z., Gergely P., Ito M., Hartshorne D.J., Erdodi F. Localization of myosin phosphatase target subunit 1 in rat brain and in primary cultures of neuronal cells. J. Comp. Neurol. 2004;478(1):72–87. PubMed

Augier E., Barbier E., Dulman R.S., Licheri V., Augier G., Domi E. A molecular mechanism for choosing alcohol over an alternative reward. Science. 2018;360(6395):1321–1326. PubMed

Caspers S., Rockner M.E., Jockwitz C., Bittner N., Teumer A., Herms S. Pathway-Specific Genetic Risk for Alzheimer's Disease Differentiates Regional Patterns of Cortical Atrophy in Older Adults. Cerebral cortex. 2019 PubMed

von Essen M.R., Sondergaard H.B., Petersen E.R.S., Sellebjerg F. IL-6, IL-12, and IL-23 STAT-Pathway Genetic Risk and Responsiveness of Lymphocytes in Patients with Multiple Sclerosis. Cells. 2019;8(3) PubMed PMC

Walsh N., Zhang H., Hyland P.L., Yang Q., Mocci E., Zhang M. Agnostic Pathway/Gene Set Analysis of Genome-Wide Association Data Identifies Associations for Pancreatic Cancer. J. Natl. Cancer Inst. 2018 PubMed PMC

van de Haar J., Canisius S., Yu M.K., Voest E.E., Wessels L.F.A., Ideker T. Identifying Epistasis in Cancer Genomes: a Delicate Affair. Cell. 2019;177(6):1375–1383. PubMed PMC

Sohail M., Vakhrusheva O.A., Sul J.H., Pulit S.L., Francioli L.C., van den Berg L.H. Negative selection in humans and fruit flies involves synergistic epistasis. Science. 2017;356(6337):539–542. PubMed PMC

Campbell R.F., McGrath P.T., Paaby A.B. Analysis of Epistasis in Natural Traits Using Model Organisms. Trends in genetics: TIG. 2018;34(11):883–898. PubMed PMC

Kirino Y., Bertsias G., Ishigatsubo Y., Mizuki N., Tugal-Tutkun I., Seyahi E. Genome-wide association analysis identifies new susceptibility loci for Behcet's disease and epistasis between HLA-B*51 and ERAP1. Nat. Genet. 2013;45(2):202–207. PubMed PMC

Syenina A., Vijaykrishna D., Gan E.S., Tan H.C., Choy M.M., Siriphanitchakorn T. Positive epistasis between viral polymerase and the 3′ untranslated region of its genome reveals the epidemiologic fitness of dengue virus. Proc Natl Acad Sci U S A. 2020;117(20):11038–11047. PubMed PMC

Kemble H., Eisenhauer C., Couce A., Chapron A., Magnan M., Gautier G. Flux, toxicity, and expression costs generate complex genetic interactions in a metabolic pathway. Sci Adv. 2020;6(23):eabb2236. PubMed PMC

Hammerling M.J., Fritz B.R., Yoesep D.J., Kim D.S., Carlson E.D., Jewett M.C. In vitro ribosome synthesis and evolution through ribosome display. Nat Commun. 2020;11(1):1108. PubMed PMC

Karageorgi M., Groen S.C., Sumbul F., Pelaez J.N., Verster K.I., Aguilar J.M. Genome editing retraces the evolution of toxin resistance in the monarch butterfly. Nature. 2019;574(7778):409–412. PubMed PMC

Trindade S., Sousa A., Xavier K.B., Dionisio F., Ferreira M.G., Gordo I. Positive epistasis drives the acquisition of multidrug resistance. PLoS Genet. 2009;5(7) PubMed PMC

Tang Y.Y., Posner M.I., Rothbart M.K., Volkow N.D. Circuitry of self-control and its role in reducing addiction. Trends Cogn. Sci. (Regul. Ed.) 2015;19(8):439–444. PubMed

Koob G.F., Volkow N.D. Neurobiology of addiction: a neurocircuitry analysis. The lancet Psychiatry. 2016;3(8):760–773. PubMed PMC

Gordon T.L., Meehan S.M., Schechter M.D. P and NP rats respond differently to the discriminative stimulus effects of nicotine. Pharmacol. Biochem. Behav. 1993;45(2):305–308. PubMed

Gordon T.L., Meehan S.M., Schechter M.D. Differential effects of nicotine but not cathinone on motor activity of P and NP rats. Pharmacol. Biochem. Behav. 1993;44(3):657–659. PubMed

Le A.D., Li Z., Funk D., Shram M., Li T.K., Shaham Y. Increased vulnerability to nicotine self-administration and relapse in alcohol-naive offspring of rats selectively bred for high alcohol intake. J Neurosci. 2006;26(6):1872–1879. PubMed PMC

Stewart R.B., Li T.K. The neurobiology of alcoholism in genetically selected rat models. Alcohol Health Res World. 1997;21(2):169–176. PubMed PMC

Moya I.M., Castaldo S.A., Van den Mooter L., Soheily S., Sansores-Garcia L., Jacobs J. Peritumoral activation of the Hippo pathway effectors YAP and TAZ suppresses liver cancer in mice. Science. 2019;366(6468):1029–1034. PubMed

Vriens K., Christen S., Parik S., Broekaert D., Yoshinaga K., Talebi A. Evidence for an alternative fatty acid desaturation pathway increasing cancer plasticity. Nature. 2019;566(7744):403–406. PubMed PMC

The dopamine transporter gene SLC6A3: multidisease risks