title: A heroin addiction severity-associated intronic single nucleotide polymorphism modulates alternative pre-mRNA splicing of the μ opioid receptor gene OPRM1 via hnRNPH interactions.
abstract: Single nucleotide polymorphisms (SNPs) in the OPRM1 gene have been associated with vulnerability to opioid dependence. The current study identifies an association of an intronic SNP (rs9479757) with the severity of heroin addiction among Han-Chinese male heroin addicts. Individual SNP analysis and haplotype-based analysis with additional SNPs in the OPRM1 locus showed that mild heroin addiction was associated with the AG genotype, whereas severe heroin addiction was associated with the GG genotype. In vitro studies such as electrophoretic mobility shift assay, minigene, siRNA, and antisense morpholino oligonucleotide studies have identified heterogeneous nuclear ribonucleoprotein H (hnRNPH) as the major binding partner for the G-containing SNP site. The G-to-A transition weakens hnRNPH binding and facilitates exon 2 skipping, leading to altered expressions of OPRM1 splice-variant mRNAs and hMOR-1 proteins. Similar changes in splicing and hMOR-1 proteins were observed in human postmortem prefrontal cortex with the AG genotype of this SNP when compared with the GG genotype. Interestingly, the altered splicing led to an increase in hMOR-1 protein levels despite decreased hMOR-1 mRNA levels, which is likely contributed by a concurrent increase in single transmembrane domain variants that have a chaperone-like function on MOR-1 protein stability. Our studies delineate the role of this SNP as a modifier of OPRM1 alternative splicing via hnRNPH interactions, and suggest a functional link between an SNP-containing splicing modifier and the severity of heroin addiction.
authors.affiliation: Department of Neurology and the Molecular Pharmacology & Chemistry Program, and.
authors.forename: Jin
authors.initials: J
authors.lastname: Xu
authors.affiliation: Department of Neurology and the Molecular Pharmacology & Chemistry Program, and.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Lu
authors.affiliation: Department of Neurology and the Molecular Pharmacology & Chemistry Program, and.
authors.forename: Mingming
authors.initials: M
authors.lastname: Xu
authors.affiliation: Department of Neurology and the Molecular Pharmacology & Chemistry Program, and.
authors.forename: Ling
authors.initials: L
authors.lastname: Pan
authors.affiliation: Department of Neurology and the Molecular Pharmacology & Chemistry Program, and.
authors.forename: Yi
authors.initials: Y
authors.lastname: Deng
authors.affiliation: Laboratory of Behavioral Neuroscience, Ningbo Addiction Research and Treatment Center, School of Medicine, Ningbo University, Ningbo, Zhejiang 315211, People's Republic of China.
authors.forename: Xiaohu
authors.initials: X
authors.lastname: Xie
authors.affiliation: Laboratory of Behavioral Neuroscience, Ningbo Addiction Research and Treatment Center, School of Medicine, Ningbo University, Ningbo, Zhejiang 315211, People's Republic of China.
authors.forename: Huifen
authors.initials: H
authors.lastname: Liu
authors.affiliation: Laboratory of Clinical Biochemistry, Ningbo No. 2 Hospital, Zhejiang 315100, People's Republic of China.
authors.forename: Shixiong
authors.initials: S
authors.lastname: Ding
authors.affiliation: Departments of Psychiatry and Neuroscience, Icahn School of Medicine at Mount Sinai, New York, New York 10029, and.
authors.forename: Yasmin L
authors.initials: YL
authors.lastname: Hurd
authors.affiliation: Department of Neurology and the Molecular Pharmacology & Chemistry Program, and.
authors.forename: Gavril W
authors.identifier: http://orcid.org/0000-0003-0060-6996
authors.initials: GW
authors.lastname: Pasternak
authors.affiliation: Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, New York 10065.
authors.forename: Robert J
authors.identifier: http://orcid.org/0000-0003-3539-5391
authors.initials: RJ
authors.lastname: Klein
authors.affiliation: Susan Lehman Cullman Laboratory for Cancer Research, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, Piscataway, New Jersey 08854.
authors.forename: Luca
authors.initials: L
authors.lastname: Cartegni
authors.affiliation: Laboratory of Behavioral Neuroscience, Ningbo Addiction Research and Treatment Center, School of Medicine, Ningbo University, Ningbo, Zhejiang 315211, People's Republic of China, pany@mskcc.org whzhou@vip.163.com.
authors.forename: Wenhua
authors.identifier: http://orcid.org/0000-0002-4804-7559
authors.initials: W
authors.lastname: Zhou
authors.affiliation: Department of Neurology and the Molecular Pharmacology & Chemistry Program, and pany@mskcc.org whzhou@vip.163.com.
authors.forename: Ying-Xian
authors.initials: YX
authors.lastname: Pan
mesh_terms: D000483:Alleles; D005787:Gene Frequency; D020022:Genetic Predisposition to Disease; D005838:Genotype; D006239:Haplotypes; D006556:Heroin Dependence / Q000235:genetics* / Q000378:metabolism; D034541:Heterogeneous-Nuclear Ribonucleoprotein Group F-H / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007438:Introns; D015810:Linkage Disequilibrium; D008297:Male; D020641:Polymorphism, Single Nucleotide; D012326:RNA Splicing; D017450:Receptors, Opioid, mu / Q000235:genetics* / Q000378:metabolism; D012720:Severity of Illness Index
chemical_list: D034541:Heterogeneous-Nuclear Ribonucleoprotein Group F-H; C485764:OPRM1 protein, human; D017450:Receptors, Opioid, mu
keywords: SNP; addiction; heroin; hnRNPH; opioid receptor; splicing