Integrative genetics of behavior with high throughput technologies

行为的综合遗传学与高通量技术

• 批准号: 8559257
• 负责人: David Goldman
• 金额: $ 310.53万
• 依托单位: NATIONAL INSTITUTE ON ALCOHOL ABUSE AND ALCOHOLISM
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8559257
• 关键词: 4-aminospiroperidol; Acute; African; African American; Alcohol dependence; Alcoholism; Alcohols; Alleles; American Indians; Animal Model; Anterior; Antipsychotic Agents; Anxiety; Behavior; Behavioral Genetics; Brain; Brain imaging; Brain-Derived Neurotrophic Factor; CRH gene; Child; Childhood; Chronic; Clinical; Clozapine; Cocaine; Cognition; Cognitive; Cognitive deficits; Complex; Corpus striatum structure; Craniocerebral Trauma; DNA Sequence; DRD2 gene; Data Set; Databases; Depressed mood; Diagnosis; Diazepam; Disease; Dopamine; Dorsal; Drug Exposure; Electroencephalography; Emotions; Environmental Risk Factor; Episodic memory; Family; Founder Generation; GTP Cyclohydrolase; Gene Cluster; Gene Expression Profile; Genes; Genetic; Genetic Polymorphism; Genetic Variation; Genomics; Genotype; HTR2A gene; Haplotypes; Histones; Human; Image; Impulsive Behavior; In Vitro; Link; Linkage Disequilibrium; Macaca mulatta; Maternal Deprivation; Measures; Mental disorders; Methaqualone; Methylation; Minisatellite Repeats; Molecular; Mus; Naltrexone; National Institute of Drug Abuse; Nature; Neurobiology; Nicotine; Nonsense-Mediated Decay; Outcome; Pain; Pain Threshold; Paper; Patients; Phenotype; Play; Population; Poverty; Preparation; Proteins; Psychiatric Diagnosis; RNA; Reporting; Risk; Risk Factors; Role; Sampling; Scanning; Schizophrenia; Selective Serotonin Reuptake Inhibitor; Sequence Analysis; Series; Single Nucleotide Polymorphism; Smoker; Stress; Substance Addiction; Suicide; Terminator Codon; Testosterone; Variant; Woman; addiction; alcohol response; anti social; chronic pain; cognitive function; dosage; early-onset alcoholism; emotional stimulus; functional genomics; gene interaction; genetic analysis; genome wide association study; genome-wide; high risk; high throughput technology; in vivo; knockout gene; neuroimaging; neuropeptide Y; next generation; nicotine craving; novel; proband; problem drinker; promoter; receptor expression; response; serotonin transporter; trait; treatment response

Identification of functional variants is an end-game of analysis of genetically influenced diseases and a starting point to understand roles of genes in behaviors. Public databases are populated with >22 million sequence variants, mostly single nucleotide polymorphisms. However, most rare and uncommon variants are unknown, and functionality of most is unknown. Through a combination of in-vivo and in vitro functional genomics studies we have discovered several functional loci and demonstrated their roles in complex behaviors relevant to alcoholism and addictions. We discovered or helped define in vitro and in vivo effects of a series of functional polymorphisms altering behavior. In alcoholism, an OPRM1 Asn40Asp missense variant (Bergen et al, 1997) was shown by others to be functional and linked to naltrexone treatment response (Anton et al, 2008). A common, functional LA->LG SNP in the serotonin transporter HTTLPR locus (Hu et al) enhanced linkage to behaviors and intermediate phenotypes. These include SSRI response of depressed patients (Hu et al, 2007), neuroimaging responses to emotion, and gene x stress interactions leading to suicidality (Roy et al, 2007). Common HTR2C Ser23Cys (Lappalainen et al) and HTR2A Asn452His alleles (Ozaki et al) were detected and shown to be functional, and linked by others to behavior, including clozapine response. In first-episode schizophrenics, we found that a functional DRD2 promoter polymorphism influences antipsychotic response (Lencz et al, 2006). We traced linkages of functional loci and haplotypes of NPY (Zhu et al, 2008), GCH1 (Tegeder et al, 2006) and DISC1 (a series of papers beginning with Hodgkinson et al, 2004) to various behaviors including emotionality, schizophrenia and clinical pain outcome. By deep sequencing we found an HTR2B Stop codon that is relatively common in Finns, absent in other populations, and associated with but not determinant for impulsive behavior. This stop codon also cosegregated with impulsive behavior in families, and the mouse gene knockout was more impulsive and higher in novelty seeking. As an example of the type of molecular studies performed with polymorphisms listed in this paragraph, the HTR2B stop codon led to variable nonsense mediated decay of the HTR2B RNA and blocked expression of the receptor (Bevilacqua et al, Nature, 2010). A low expression Neuropeptide Y (NPY) haplotype increased anxiety and emotionality but had stronger effects on molecules (NPY RNA and protein) and intermediate phenotypes (brain imaging measures of responses to emotional stimuli and pain/stress) than complex behavior. (Zhou et al, Nature, 2008). Intermediate phenotypes augment genetic analyses of behavior. The imaging genetics paradigm we helped initiate (Heinz et al, 2000; Hariri et al, 2002; Egan et al, 2001; 2003; Zubieta et al, 2003) has continued to yield groundbreaking findings. With collaborators at NIDA, we helped discover that the mechanism of action of CHRNA5 Asn398, the major functional locus from nicotine GWAS, involves weakening of a Dorsal Anterior Cingulate/Ventral Striatal circuit, the weakness of which predicts nicotine craving in smokers with and without other psychiatric diagnoses (Hong et al, 2010). Clinical subphenotyping enabled linkage of HTR1B to antisocial alcoholism (Lappalainen et al), serotonin transporter (SLCA4) to anxiety (Mazzanti et al; Hariri et al), BDNF Val66Met to episodic memory (Egan et al), COMT Val158Met to anxiety (Enoch et al), executive cognition (Egan et al; Lipsky et al; Malhotra et al), and pain threshold (Zubieta et al; Diatchenko et al), and GTP cyclohydrolase to chronic pain and experimental pain response (Tegeder et al). In these studies brain imaging and cognitive measures play prominent roles. Frontal cognitive deficit is a risk factor in schizophrenia, alcoholism and other diseases. Dopamine generally enhances prefrontal cortical efficiency. Met158, a common COMT variant, leads to four-fold reduction in COMT activity. It is thus a candidate allele for cognitive function via effect on frontal dopamine. We found an allele-dosage relationship of Met158 to frontal cognitive function and diminished frontal cortical efficiency (Egan et al). The relationship to cognition is observed in populations differing in baseline cognitive function: schizophrenia, moderate-severe head injury (Lipsky et al), & controls (Malhotra et al). We proposed that Val158 has a counter-advantage: stress resiliency. In two populations Met158 predicted anxiety in women and decreased frontal EEG coherence (Enoch et al), and Met158 was associated with lower resiliency to pain/stress (Zubieta et al; Diatchenko et al). Met158 predicted inability to activate endomorphin release after pain/stress (Zubieta et al). Overall, effect sizes of genes in intermediate phenotypes is larger (Goldman and Ducci, 2007) and as illustrated by effects of genes such as NPY (Zhou et al, 2008), COMT (Zubieta et al, 2003) and CHRNA5 (Hong et al, 2010) on intermediate phenotypes. Linkage, gwas and RNA-seq are representative of genomic, hypothesis-free approaches. Linkage disequilibrium of alcoholism to GABRA2 was found at the Chr 4 GABAA subunit cluster we implicated by family linkage (Long et al). We showed the association was anxiety- modulated (Enoch et al). Another GABAA gene cluster implicated in alcoholism and alcohol response is located on Chr 5. Within this cluster we reported linkage disequilibrium to alcoholism (Radel et al) and that GABRA6 has a missense variant associated with alcohol dependence and response to alcohol and diazepam (Iwata et al, Schuckit et al). A family linkage scan yielded genome-wide significant linkage of CRH-BP (a stress-related protein) to an alcoholism-associated EEG trait and this was followed by association of CRH-BP to EEG in two populations. (Enoch et al). Our gwas of EEG (Hodgkinson et al, PNAS, 2010) in a relatively small sample of Plains Indians detected three independent genome-wide significant loci, and we replicated two loci (including one locus that was just sub-threshold) in a second population (Hodgkinson et al, PNAS 2010). We recently used next-generation sequencing to analyze brain histone methylation and transcriptome changes resulting from early maternal deprivation in Rhesus macaques (Barr et al, in preparation) and effects of chronic cocaine and alcohol in human brain (Zhou et al, PNAS 2010). The latter study found distinct differences between the gene networks that are regulated by chronic drug exposure in people, as compared to those detected in acute and subacute exposures in animal models. Founder populations, extreme probands, and exposed populations enhance power to detect gene effects. Our Finnish dataset is derived from a founder population and ascertained from criminal alcoholic probands & thus enriched for Type II early onset alcoholism. SW and Plains Indian samples represent founder populations. An African American substance dependence sample with high adversity exposure revealed GxE of childhood adversity and HTTLPR in suicidality (Roy et al, 2007). Stress and poverty, but not African ancestry, predicted high risk of addictions (Ducci et al, 2009). An MAOA VNTR previously linked to dyscontrol via stress interaction was linked to outcomes of alcohol dependence and ASPD in American Indian women, of whom half had been sexually abused as children (Ducci et al, 2008). A strong interaction between the MAOA VNTR and testosterone was observed in the Finnish criminal sample (Sjoberg et al). As mentioned, deep sequencing in Finns detected a population-specific HTR2B Stop codon significant for impulsive behavior (Bevilacqua et al, Nature, 2010).

功能变体的识别是分析受遗传影响疾病的终点，也是了解基因在行为中的作用的起点。公共数据库中有> 2200万个序列变体，主要是单核苷酸多态性。但是，最罕见和罕见的变体是未知的，大多数的功能尚不清楚。通过体内和体外功能基因组学研究的结合，我们发现了几个功能基因座，并证明了它们在与酒精中毒和成瘾有关的复杂行为中的作用。 我们发现或帮助定义了一系列功能多态性改变行为的体外和体内效应。在酒精中毒中，其他人证明了OPRM1 ASN40ASP错义变体（Bergen等，1997）具有功能性，并与纳曲酮治疗反应相关（Anton等，2008）。 5-羟色胺转运蛋白HTTLPR基因座（Hu等人）中常见的功能性LA-> lg SNP增强了与行为和中间表型的联系。其中包括抑郁患者的SSRI反应（Hu等，2007），对情绪的神经影像反应以及导致自杀性的Gene X压力相互作用（Roy等，2007）。 检测并证明了常见的HTR2C Ser23Cys（Lappalainen等人）和Htr2a ASN452HIS等位基因（Ozaki等人），并证明其功能性，并由其他人与行为联系起来，包括氯氮平反应。在第一期精神分裂症患者中，我们发现功能性DRD2启动子多态性会影响抗精神病药反应（Lencz等，2006）。我们追踪了NPY功能基因座和单倍型的联系（Zhu等，2008），GCH1（Tegeder等，2006）和Disc1（一系列从Hodgkinson等人开始的论文），包括各种行为，包括情绪，精神分裂症和临床疼痛结果。通过深入测序，我们发现了一个HTR2B终止密码子，该密码子在Finn中相对常见，在其他人群中不存在，并且与冲动行为相关但不确定性。这个停止的密码子也与家庭冲动的行为进行了混合，而小鼠基因敲除在寻找新颖性中更加冲动和更高。作为本段中列出的多态性进行的分子研究类型的一个例子，HTR2B终止密码子导致HTR2B RNA的无义介导的衰减变化并阻断受体的表达（Bevilacuququa等人，自然，2010年）。 低表达神经肽Y（NPY）单倍型增加了焦虑和情绪性，但对分子（NPY RNA和蛋白质）以及中间表型（脑成像对情绪刺激和疼痛/压力的反应测量）的影响比复杂的行为更强。 （Zhou等人，自然，2008年）。 中间表型增强了行为的遗传分析。我们帮助启动的成像遗传学范式（Heinz等，2000； Hariri等，2002； Egan等，2001； 2003； Zubieta等，2003）继续产生突破性的发现。与NIDA的合作者一起，我们帮助发现ChRNA5 ASN398的作用机理是Nicotine GWAS的主要功能基因座，涉及削弱背扣带回/腹侧纹状体电路的弱点，其弱点预测其尼古丁在吸烟者中渴望吸烟的尼古丁在烟雾中渴望与其他精神病学诊断（Hong extress extorsise（Hong et al ang ang et al，Hong et al and e al and Allosise）。临床亚近代分型启用HTR1B与反社会酒精中毒（Lappalainen等），5-羟色胺转运蛋白（SLCA4）与焦虑（Mazzanti等； Hariri等），BDNF Val66met，comt val158met to焦虑（En）ENCERKY（ENCEN）ETICETION（EN）ET;等）和疼痛阈值（Zubieta等人； Diatchenko等）和GTP环氢酶对慢性疼痛和实验性疼痛反应（Tegeder等人）。在这些研究中，大脑成像和认知措施扮演着重要角色。额叶认知缺陷是精神分裂症，酒精中毒和其他疾病的危险因素。多巴胺通常提高额叶皮质效率。 MET158是一种常见的COMT变体，可导致COMT活动减少四倍。因此，它是通过对额叶多巴胺的作用来实现认知功能的候选等位基因。我们发现Met158与额叶认知功能的等位基因剂量关系，并降低了额叶皮质效率（Egan等人）。在基线认知功能不同的人群中观察到与认知的关系：精神分裂症，中度重度头部损伤（Lipsky等人）和对照组（Malhotra等人）。我们提出Val158具有相反的优势：应力弹性。在两个人群中，MET158预测了女性的焦虑，额头相干性降低（Enoch等人），而Met158与对疼痛/压力的弹性降低有关（Zubieta等人； Diatchenko等）。 MET158预测无法激活疼痛/应激后的内啡肽释放（Zubieta等人）。总体而言，中间表型中基因的效应大小较大（Goldman和Ducci，2007），如NPY（Zhou等，2008），COMT（Zubieta等，2003）和Chrna5（Hong等人，2010年）对中间型型的效果所说明。 链接，GWAS和RNA-SEQ代表了基因组无假设方法。酒精中毒与gabra2的连锁不平衡是在我们与家庭联系（Long等人）牵连的Chr 4 Gabaa亚基群集中发现的。我们表明这种关联是焦虑调节的（Enoch等人）。涉及酒精中毒和酒精反应的另一个GABAA基因簇位于Chr 5中。在此集群中，我们报告了与酒精中毒（Radel等人）的连锁不平衡，并且Gabra6具有与酒精依赖和对酒精和地致致致变的反应有关的错义变体（Iwata等人（Iwata等，Schuckit等）。家庭连锁扫描产生了CRH-BP（与应激相关的蛋白）与酒精中毒相关的脑电图性状的明显连接，随之而来的是CRH-BP与两个人群中的EEG相关。 （Enoch等人）。我们的脑电图（Hodgkinson等人，PNA，2010年）在相对较小的平原样本中，印第安人检测到了三个独立的全基因组显着的基因座，我们在第二个人群中复制了两个基因座（包括一个仅次于阈值的基因座）（Hodgkinson等人，PNAS，PNAS 2010）。最近，我们使用下一代测序来分析恒河猕猴早期孕产妇剥夺引起的脑组蛋白甲基化和转录组变化（Barr等人的制备中）以及人脑中慢性可卡因和酒精的影响（Zhou等，PNAS，2010年）。后一项研究发现，与动物模型中急性和亚急性暴露的基因网络受到慢性药物暴露调节的基因网络之间的明显差异。 创始人人群，极端概率和暴露种群增强了检测基因效应的能力。我们的芬兰数据集源自创始人人口，并从犯罪的酒精探险中确定，因此富含II型早期发作的酒精中毒。 SW和Plains印度样品代表创始人人群。具有高逆性暴露的非裔美国物质依赖性样本显示，自杀性的儿童逆境和HTTLPR的GXE（Roy等，2007）。压力和贫穷，但不是非洲血统，预测了成瘾的高风险（Ducci等，2009）。以前通过压力相互作用与Dyscontrol相关的MAOA VNTR与美洲印第安妇女的酒精依赖和ASPD的结果有关，其中一半是儿童性虐待的（Ducci等，2008）。在芬兰的犯罪样本（Sjoberg等）中观察到了MAOA VNTR和睾丸激素之间的强烈相互作用。如前所述，芬兰人中的深层测序检测到人群特异性的HTR2B停止密码子对于冲动行为而言重要（Bevilacqua等，Nature，2010年）。