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: 9559246
• 负责人: David Goldman
• 金额: $ 415.25万
• 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/9559246
• 关键词: 4-aminospiroperidol; African American; Alcohol dependence; Alcoholism; Alcohols; Alleles; American Indians; Animal Model; Animals; Anterior; Antipsychotic Agents; Anxiety; Autopsy; Behavior; Behavioral; Behavioral Genetics; Brain; Brain imaging; Brain-Derived Neurotrophic Factor; CRH gene; Cell model; Cells; Child; Clinical; Clozapine; Cognition; Cognitive deficits; Collection; Complex; Corpus striatum structure; Craniocerebral Trauma; DISC1 gene; DRD2 gene; Data Set; Development; Diagnosis; Diazepam; Disease; Dopamine; Dorsal; Drosophila genus; Electroencephalography; Emotional; Emotions; Endocrine; Environmental Risk Factor; Epigenetic Process; Episodic memory; Exposure to; Extracellular Domain; Family; Founder Generation; GTP Cyclohydrolase; Gene Cluster; Genes; Genetic; Genetic Polymorphism; Genetic Variation; Genetic study; Genomic approach; Genomics; Genotype; Glutamate Receptor; Glutamates; Goals; Grant; HTR2A gene; Haplotypes; Heritability; Human; Impulsive Behavior; Impulsivity; Investigation; Knowledge; Ligands; Link; Massive Parallel Sequencing; Mental disorders; Methaqualone; Minisatellite Repeats; Mus; Naltrexone; National Institute of Drug Abuse; Nature; Neurobiology; Neuroglia; Neurosciences; Nicotine; Nicotine Dependence; Nonsense-Mediated Decay; Outcome; Pain; Phenotype; Placebo Effect; Population; Poverty; Proteins; RNA; Rattus; Risk; Risk Factors; Rivers; Role; Sampling; Scanning; Schizophrenia; Science; Sexual abuse; Source; Stress; Study models; Substance Addiction; Suicide; Terminator Codon; Testosterone; Translational Research; Validation; Variant; Wistar Rats; Withdrawal; Woman; addiction; alcohol preferring rats; alcohol response; alcohol use disorder; anti social; base; cannabinoid receptor; chronic pain; cognitive function; comparative genomics; craving; deep sequencing; dosage; drug reward; drug withdrawal; exome; exome sequencing; externalizing behavior; fatty acid amide hydrolase; functional genomics; gene discovery; gene interaction; genome wide association study; genome-wide; genome-wide analysis; high risk; high throughput technology; imaging genetics; induced pluripotent stem cell; knockout gene; metabotropic glutamate receptor 2; neuroimaging; neuropeptide Y; novel; pediatric trauma; preference; premenstrual dysphoric disorder; proband; problem drinker; promoter; receptor expression; resilience; response; risk variant; serotonin transporter; stress resilience; trait; transcriptomics; treatment response

Identification of functional variants is an end-game of analysis of genetically influenced diseases and starting point to understand roles of genes. Addictions are moderately to highly heritable but most of the genetic variance is unexplained by functional loci or replicated linkages. Using genomic approaches and focused studies on implicated genes, we have discovered several functional loci with roles in complex behaviors relevant to alcohol use disorders (AUD) and other addictions. These studies make use of founder populations, people at risk due to exposures, and model organisms. We use intermediate phenotypes that are closer to gene action. Model organisms, including artificially selected mice, rats and fruitflies are used for gene discovery, as well as validation and we use cellular models including iPSC's to bridge between animal models and humans. An OPRM1 Asn40Asp missense variant (Bergen et al, 1997) was found to be functional and linked to naltrexone treatment response in AUD(Anton et al, 2008). A common, functional SNP in the serotonin transporter HTTLPR (Hu et al, 2007) enhanced linkage to OCD,neuroimaging responses to emotion, and gene x stress interactions leading to suicidality (Roy et al, 2007). Common HTR2C Ser23Cys (Lappalainen et al, 1999, Okada et al, 2004) and HTR2A Asn452His variants(Ozaki et al, 1997) were found, shown to be functional, and linked to behavior, including clozapine response. In first-episode schizophrenics, we found that a functional DRD2 promoter variant 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 (Hodgkinson et al, 2004) to emotionality, schizophrenia and pain. By deep sequencing we found an HTR2B stop codon that is relatively common in Finns, a founder population, absent in others, and associated with but not determinant for impulsive behavior. This stop codon cosegregated with ASPD and AUD in families and the mouse gene knockout was impulsive and high in novelty seeking and novelty response. It leads to variable nonsense mediated decay of the HTR2B RNA and blocked HT2B receptor expression (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 responses to emotion and pain/stress)(Zhou et al, Nature, 2008). Because intermediate phenotypes augment analysis of behavior (Ducci and Goldman)we proposed a multidimensional Addictions Neuroclinical Assessment based on the addiction cycle (Kwako et al, Biol Psych, 2016). The imaging genetics paradigm we helped initiate (Heinz et al, 2000; Hariri et al, 2002; Egan et al, 2001; 2003; Zubieta et al, 2003) led to groundbreaking findings. We found a mechanism of CHRNA5 Asn398, a functional locus altering nicotine addiction risk. The risk allele weakens brain connectivities including a Dorsal Anterior Cingulate/Ventral Striatal circuit predictive of craving (Hong et al, 2010). Clinical subphenotyping enabled linkage of HTR1B to antisocial alcoholism (Lappalainen et al, 1998), serotonin transporter (SLCA4) to anxiety (Mazzanti et al; Hariri et al), BDNF Val66Met to episodic memory (Egan et al, cell, 2003), and GTP cyclohydrolase to chronic pain and experimental pain response (Tegeder et al, 2006). 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 cortical efficiency (Egan et al, 2001). The effect on cognition is seen in populations differing in baseline cognitive function: schizophrenia, head injury (Lipsky et al, 2005, 2011), & controls (Malhotra et al, 2002, 2009). 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, 2003), and Met158 was associated with lower resiliency to pain/stress (Zubieta et al, Science, 2003; Diatchenko et al, 2005, 2006). We have extended knowledge of GxE effects of stress related genes such as COMT, NPY, SLC6A4 and SLC6A4 in many studies using both behavioral and endocrine outcomes (Lovallo et al 2015, 2016). A functional polymorphism of Fatty Acid Amide Hydrolase (FAAH) which metabolizes the endogenous ligand for cannabinoid receptors, specifically predicts placebo response to pain (Pecina et al, 2014). GWAS, transcriptomics and exome sequencing are global approaches that enable the genomic space to be searched independent of prior hypotheses. Via transcriptomics we have recently advanced understanding of glia in addictions (De Biase, 2017), developmental effects of nicotine (Jung, 2016), and found an intrinsic cellular difference in women with Premenstrual Dysphoric Disorder (Dubey,2016). The Chr 4 GABAA genecluster was implicated in alcoholism by family linkage (Long et al, 1998), an effect that appears anxiety-modulated (Enoch et al, 2006). Another GABAA gene cluster implicated in alcoholism and alcohol response is located on Chr 5 (Radel et al, 2005). GABRA6 has a missense variant associated with alcohol dependence and response to alcohol and diazepam (Iwata et al, 2005). A family linkage scan yielded genome-wide significant linkage of CRH-BP to an alcoholism-associated EEG trait and this was supported by association in two populations (Enoch et al, 2008). GWAS of EEG (Hodgkinson et al, PNAS, 2010) in Plains Indians detected three independent genome-wide significant loci(Hodgkinson et al, PNAS 2010). Via GWAS, a schizophrenia risk locus was identified in multiple populations (Lencz et al, Nat Commun, 2013). Founder, phenotypically extreme, and exposed populations enhance power to detect gene effects. Our Finnish dataset, derived from a founder population, was ascertained via criminal alcoholic probands. SW and Plains Indian samples represent founder populations. An African American substance dependence sample with high adversity revealed GxE of childhood trauma 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 a MAOA VNTR and testosterone was observed in the Finnish criminal sample (Sjoberg et al, 2008). As mentioned, deep sequencing in Finns detected a population-specific HTR2B Stop codon significant for impulsivity (Bevilacqua et al, Nature, 2010). We exome-sequenced an artificially selected animal model for alcoholism, the P/NP rat, and discovered a stop codon in the metabotropic glutamate receptor 2 (Grm2) gene that leads to altered glutamate function and increased alcohol preference in alcohol-preferring rats (Zhou et al, 2013). The Grm2 Stop codon is common (0.08) in parental Charles River Wistar rats, and fixed in Wistar from some sources. It leads to uncompensated changes in glutamate function. It was genetically fixed by artificial selection in the alcohol preferring and non-preferring rats illustrating the power of using such strains to discover alleles that alter behavior. Via parallel exome sequencing in selected strains and people we are identifying genes involved in drug reward and withdrawal. As facilitated by a NIDA RFA leading to two U01 grants we have identified new loci altering addiction in rat and Drosophila.

功能变异的鉴定是遗传影响疾病分析的最终阶段，也是理解基因作用的起点。成瘾具有中度至高度遗传性，但大多数遗传变异无法通过功能基因座或复制连锁来解释。利用基因组方法和对相关基因的集中研究，我们发现了几个在与酒精使用障碍 (AUD) 和其他成瘾相关的复杂行为中发挥作用的功能位点。这些研究利用了创始人群体、因暴露而面临风险的人以及模型生物。我们使用更接近基因作用的中间表型。模型生物体，包括人工选择的小鼠、大鼠和果蝇，用于基因发现和验证，我们使用包括 iPSC 在内的细胞模型来连接动物模型和人类。 OPRM1 Asn40Asp 错义变体（Bergen 等人，1997）被发现具有功能并与 AUD 中的纳曲酮治疗反应相关（Anton 等人，2008）。血清素转运蛋白 HTTLPR 中常见的功能性 SNP（Hu 等，2007）增强了与强迫症、神经影像对情绪的反应以及导致自杀的基因 x 应激相互作用的联系（Roy 等，2007）。 发现了常见的 HTR2C Ser23Cys（Lappalainen 等人，1999；Okada 等人，2004）和 HTR2A Asn452His 变体（Ozaki 等人，1997），它们具有功能性，并且与行为（包括氯氮平反应）相关。在首发精神分裂症患者中，我们发现功能性 DRD2 启动子变异会影响抗精神病药物反应（Lencz 等，2006）。我们追踪了 NPY（Zhu 等人，2008）、GCH1（Tegeder 等人，2006）和 DISC1（Hodgkinson 等人，2004）的功能位点和单倍型与情绪、精神分裂症和疼痛的联系。通过深度测序，我们发现了一个 HTR2B 终止密码子，该密码子在芬兰人（一个创始人群体）中相对常见，但在其他人群中不存在，与冲动行为相关但不是决定因素。该终止密码子与 ASPD 和 AUD 在家族中共分离，小鼠基因敲除具有冲动性，新颖性寻求和新颖性反应较高。它导致 HTR2B RNA 的可变无义介导的衰变并阻断 HT2B 受体表达（Bevilacqua 等人，Nature，2010）。 低表达神经肽 Y (NPY) 单倍型会增加焦虑和情绪，但对分子（NPY RNA 和蛋白质）和中间表型（对情绪和疼痛/压力的大脑成像反应）有更强的影响（Zhou 等人，Nature，2008）。 由于中间表型增强了行为分析（Ducci 和 Goldman），我们提出了基于成瘾周期的多维成瘾神经临床评估（Kwako 等人，Biol Psych，2016）。我们帮助发起的成像遗传学范式（Heinz 等，2000；Hariri 等，2002；Egan 等，2001；2003；Zubieta 等，2003）带来了突破性的发现。我们发现了 CHRNA5 Asn398 的机制，这是一个改变尼古丁成瘾风险的功能位点。风险等位基因会削弱大脑连接，包括预测渴望的背侧前扣带回/腹侧纹状体回路（Hong 等，2010）。临床亚表型分析使 HTR1B 与反社会性酒精中毒（Lappalainen 等人，1998）、血清素转运蛋白（SLCA4）与焦虑（Mazzanti 等人；Hariri 等人）、BDNF Val66Met 与情景记忆（Egan 等人，cell，2003）以及 GTP 环化水解酶与慢性疼痛和实验性疼痛反应（Tegeder 等人， 2006）。额叶认知缺陷是精神分裂症、酗酒和其他疾病的危险因素。多巴胺通常会增强前额皮质的效率。 Met158 是一种常见的 COMT 变体，可导致 COMT 活性降低四倍。因此，它是通过影响额叶多巴胺而发挥认知功能的候选等位基因。我们发现 Met158 与额叶认知功能和皮质效率降低存在等位基因剂量关系（Egan 等，2001）。对认知的影响在基线认知功能不同的人群中可见：精神分裂症、头部损伤（Lipsky 等，2005、2011）和对照（Malhotra 等，2002、2009）。我们提出 Val158 有一个反优势：压力恢复能力。在两个人群中，Met158 预测女性焦虑并降低额叶脑电图一致性（Enoch 等，2003），并且 Met158 与较低的疼痛/压力弹性相关（Zubieta 等，Science，2003；Diatchenko 等，2005，2006）。在许多使用行为和内分泌结果的研究中，我们扩展了对压力相关基因（例如 COMT、NPY、SLC6A4 和 SLC6A4）的 GxE 影响的了解（Lovallo 等人 2015，2016）。 脂肪酸酰胺水解酶 (FAAH) 的功能多态性代谢大麻素受体的内源配体，可特异性预测安慰剂对疼痛的反应 (Pecina 等人，2014)。 GWAS、转录组学和外显子组测序是全局方法，使基因组空间的搜索能够独立于先前的假设。通过转录组学，我们最近对成瘾中的神经胶质细胞（De Biase，2017）、尼古丁的发育影响（Jung，2016）有了更深入的了解，并发现患有经前焦虑症的女性存在内在的细胞差异（Dubey，2016）。 Chr 4 GABAA 基因簇通过家庭联系与酗酒有关（Long 等，1998），这种效应似乎是焦虑调节的（Enoch 等，2006）。另一个与酗酒和酒精反应有关的 GABAA 基因簇位于 Chr 5（Radel 等，2005）。 GABRA6 具有与酒精依赖以及对酒精和地西泮的反应相关的错义变体（Iwata 等，2005）。家族连锁扫描发现 CRH-BP 与酗酒相关的脑电图特征在全基因组范围内显着关联，并且这得到了两个人群关联的支持（Enoch 等，2008）。平原印第安人脑电图的 GWAS（Hodgkinson 等人，PNAS，2010）检测到三个独立的全基因组显着位点（Hodgkinson 等人，PNAS 2010）。通过 GWAS，在多个人群中确定了精神分裂症风险位点（Lencz 等人，Nat Commun，2013）。 创始人、表型极端和暴露人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群体，是通过酗酒犯罪先证者确定的。西南和平原印第安人样本代表了创始人群体。一个处于高度逆境的非裔美国人物质依赖样本揭示了童年创伤的 GxE 和自杀倾向的 HTTLPR（Roy 等，2007）。压力和贫困，而非非洲血统，预示着成瘾的高风险（Ducci et al, 2009）。 MAOA VNTR 先前通过压力相互作用与失控有关，但与美国印第安妇女的酒精依赖和 ASPD 的结果有关，其中一半人在儿童时期遭受过性虐待（Ducci 等，2008）。在芬兰犯罪样本中观察到 MAOA VNTR 和睾酮之间存在强烈的相互作用（Sjoberg 等，2008）。如前所述，芬兰人的深度测序检测到了一个对冲动具有重要意义的群体特异性 HTR2B 终止密码子（Bevilacqua 等人，Nature，2010）。我们对人工选择的酗酒动物模型 P/NP 大鼠进行了外显子组测序，并在代谢型谷氨酸受体 2 (Grm2) 基因中发现了一个终止密码子，该终止密码子会导致嗜酒大鼠的谷氨酸功能改变和酒精偏好增加 (Zhou et al, 2013)。 Grm2 终止密码子在亲代 Charles River Wistar 大鼠中很常见 (0.08)，并通过某些来源固定在 Wistar 中。它会导致谷氨酸功能发生无补偿的变化。它是通过人工选择在偏好酒精和不偏好酒精的老鼠中进行遗传固定的，这说明了使用此类菌株来发现改变行为的等位基因的力量。通过对选定菌株和人群进行平行外显子组测序，我们正在识别与药物奖励和戒断有关的基因。在获得两项 U01 资助的 NIDA RFA 的推动下，我们在大鼠和果蝇中发现了改变成瘾的新位点。