Comparative Analysis of Genetics of Stress Response and Reward

应激反应和奖励的遗传学比较分析

基本信息

批准号：
10267514
负责人：
Christina Barr
金额：
$ 148.59万
依托单位：
NATIONAL INSTITUTE ON ALCOHOL ABUSE AND ALCOHOLISM
依托单位国家：
美国
项目类别：
财政年份：
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/10267514
关键词：
Alcohol abuse Alcohol consumption Alcohol dependence Alcohol or Other Drugs use Alcohols Alleles Alzheimer&apos s Disease Animals Antiemetics Anxiety Archives Behavior Behavioral Bioavailable Brain Brain region Canis familiaris Collaborations Consumption Corticotropin Cues DNA Data Set Disease Domestic Animals Empathy Environmental Exposure Environmental Risk Factor Epigenetic Process Equilibrium Ethanol Evolution Failure Family Felidae Fetal Alcohol Exposure Genes Genetic Genetic Polymorphism Genetic Variation Genetic study Genomic approach Genomics Genotype Home environment Human Human Genetics Hypothalamic structure Immersion Impulsivity Individual Individual Differences Label Laboratories Learning Light Link Macaca Macaca mulatta Mental disorders Mission Modeling Monkeys Mothers Mutation National Institute on Alcohol Abuse and Alcoholism Neurobiology Neurodevelopmental Disorder Neurons Nociception Nurseries ORL1 receptor Ondansetron Opioid Receptor Oral Pain Pan Genus Pathology Pathway interactions Personality Disorders Pharmacogenetics Phenotype Population Pregnancy Primates Psychopathology RNA Recording of previous events Reporting Rewards Rhesus Risk Risk Factors Rodent Role Sampling Self Administration Sensory Serotonergic System Serotonin Spinal Ganglia Stress System Tactile Temperament Terminator Codon Tissues Touch sensation United States National Institutes of Health Variant Water Withdrawal Woman addiction alcohol abuse therapy alcohol exposure alcohol seeking behavior alcohol use disorder anxious autism spectrum disorder behavioral genomics behavioral phenotyping biological adaptation to stress brain tissue comorbidity comparative conditioned place preference dosage early experience early pregnancy exome sequencing frontal lobe genetic analysis genetic risk factor genetic variant genome sequencing in utero individual variation loss of function mu opioid receptors next generation sequencing nociceptin nonhuman primate peer preference prenatal exposure receptor response sample fixation social social cognition tissue stress trait transcriptome sequencing

项目摘要

With the expansion in capacity of Next Generation Sequencing for performing and refining genome sequencing, we have begun to uncover genetic and environmental factors in the nonhuman primate that contribute to individual variation in traits observed among various human neurodevelopmental and psychiatric disorders, including the addictions. Using archived datasets and relying on ongoing collaborations, we have been deriving addiction-related traits from reasonably sized datasets for examination of genotype-phenotype correlations in environmentally controlled populations. We postulate potential pathways through which the human alcohol use disorders occur, several of which are discussed below. We performed Whole Exome Sequencing (WES) on a population of macaques that had been exposed to alcohol in utero. Using DNA from individuals with extreme FAE phenotypes (high/agitated vs. calm/mellow temperament, high vs. low separation induced ACTH response, best vs. worst trials to criterion for delayed nonmatching to sample, and high/defensive vs. low/nonreactive for sensory processing), we found stop codon (missense) variants in genes influencing neuron and stress system function, including that encoding the D subunit for the serotonin HT-3 receptor (rhHTR3D). Given the role of the serotonin system in anxiety, sensory processing and alcohol-seeking behavior in addition to the fact that the HT-3 receptor antagonist, ondansetron, has not only been proposed for the off-label treatment of alcohol dependence but is also commonly used as an anti-emetic for women during pregnancy, HTR3D seemed like an interesting candidate to consider within the context of this model. Thus far, the HTR3D gene has only been characterized in humans, chimpanzees, and dogs. In each of these species, the HTRD3 gene exists in a cluster with HTR3C and HTR3E and appears to be largely confined in its expression to the dorsal root ganglion (DRG), although there are low levels of expression observed in other neuronal tissues, including frontal lobe and the hypothalamus. The high level of expression in the DRG is interesting in that 70% of the animals we screened were high sensory defensive if they were carriers of the alternative rhHTR3D allele. As our initial screen suggested the possibility that HTR3D variation may have a role in sensory processing, we wanted to examine whether individual differences in sensory sensitivity varied according to prenatal ethanol exposure and genotype. We found interactions among rhHTR3D genotype and timing of prenatal exposure. Genotype appeared to predict mean sensory magnitude (withdrawal from tactile stimulation and failure to habituate to light touch) in animals that were divergent for early alcohol exposure, but among late gestation exposed animals, there were qualitative interactive effects according to whether mothers consumed alcohol during early pregnancy. Although the sample is small, there is a scarcity of studies reporting genetic risk factors for FAEs, and very little is known about the HTR3D gene, making our findings intriguing. This gene appears to be absent in rodents, which speaks to the tremendous translational value of studies performed in primates. Whether there are HTR3D mutations in human alcohol-dependent samples that might be important in predicting ondansetron response seems plausible. Here, we find that there is an allele dosage effect for the prediction of individual differences in voluntary ethanol self- administration in the macaque. As stated above, HT-3 receptor antagonism has been used to treat alcohol addiction. A stop codon variant could, in effect, behave as an antagonist for this receptor. As one might expect, this loss of function variant is associated with lower levels of self- administration. Perhaps the most valuable feature of the NIH rhesus population (maintained decades ago) is that at least 12 of the individuals assessed were reared in a nursery (PR, peer reared). Monkeys reared under these conditions often develop to be more anxious, impulsive or aggressive and have social cognition differences as well as features of attachment disorders. When given a choice, they also voluntarily consume more alcohol. Because of the early rearing paradigm, we are provided the opportunity to examine G x E interactions. At minimum, datasets archived from this colony provide access to behavioral phenotypes for a population enriched for individuals demonstrating behavioral pathology due to their early environmental exposures. This may increase sensitivity for performing genetics studies to model the trajectories through which humans develop psychopathology and alcohol problems. RNA-Seq of brain tissue from stress exposed animals may be especially useful for identifying variation within functionally important genes. By sequencing RNA from samples archived from macaque brains from individuals divergent for early rearing history, we can screen for variation within genes expressed in behaviorally-relevant and stress-responsive brain regions. Among the SNPs detected in rhesus macaques using RNA-Seq was an nsSNP in the Nociceptin/Orphanin FQ (rhOPRL1, also known as kappa 3) receptor (rhOPRL1 His243Arg). As with other opioid receptors, the Nociceptin/Orphanin FQ receptor confers differential sensitivity to pain in addition to phenotypes relating to stress and substance use. The rhOPRL1 His243Arg SNP is located in a region highly conserved among this class of receptors, and polyphen predicted this SNP to be Damaging (Score of 0.999). There is evidence that functionally important OPRL1 variation occurs across species. In humans, expression of OPRL1 is altered in autism spectrum disorders and Alzheimers Disease, and in dogs, ancient missense polymorphisms have gone to fixation in certain domestic dog breeds (notably water retrievers, selectively bred to withstand immersion in cold water) a phenotype that could be considered in the context of the human cold pressor/pain response. Studies performed in laboratory rodents demonstrate that ethanol self-administration in strains selected for alcohol preference can be inhibited with administration of a Nociceptin receptor antagonist. This suggests that genetic variation at this receptor could contribute to the alcohol preference phenotype. These studies show that antagonism of the receptor not only blocks acquisition of conditioned place preference, but also the rate to transitioning to the dependent state with intermittent access; Also blocked are stress- and cue-induced reinstatement in post-dependent animals. For these reasons, antagonism of this receptor has been proposed as a potential treatment for alcohol addictions. There is a dearth of genetic studies investigating a role for naturally occurring genetic variation at OPRL1 in relation to addiction-related phenotypes and the pharmacogenetics of receptor antagonists. We show that damaging variation at rhOPRL1 predicts individual differences in early acquisition of alcohol intake in rhesus macaques. Although there were no interactions, inclusion of early experience in the model appeared to be important for this sample. It is conceivable that OPRL1 genotype effects could be more appreciable in human populations with a high prior stress burden. With access to an orally bioavailable nociception receptor antagonist, proposed as a potential treatment for MDD and Alcohol Use Disorders, studies aimed at examination of a pharmacogenetic effect at OPRL1 may also be possible in humans. As stated above, there are functional Mu-Opioid Receptor variants in both humans and macaques. Given the link between variation in these genes in predicting differences in sensitivity to the pain of social rejection and empathic response, variation at both genes may also relate to risk for personality disorders known to be co-morbid with AUDs.

随着下一代测序执行和完善基因组测序的能力的扩展，我们已经开始发现非人类灵长类动物中的遗传和环境因素，这些因素导致了在各种人类神经发育和精神疾病（包括成瘾）中观察到的特征的个体差异。利用存档的数据集并依靠持续的合作，我们一直在从合理大小的数据集中导出与成瘾相关的特征，以检查环境控制人群中的基因型-表型相关性。我们假设人类酒精使用障碍发生的潜在途径，下面讨论其中的一些途径。我们对一群在子宫内接触过酒精的猕猴进行了全外显子组测序 (WES)。使用具有极端 FAE 表型的个体的 DNA（高/激动与平静/温和气质、高与低分离诱导的 ACTH 反应、最佳与最差试验来确定延迟与样本不匹配的标准，以及高/防御与低/非反应性）感觉处理），我们在影响神经元和应激系统功能的基因中发现了终止密码子（错义）变异，包括编码血清素 HT-3 受体 D 亚基的基因(rhHTR3D)。鉴于 5-羟色胺系统在焦虑、感觉处理和酗酒行为中的作用，以及 HT-3 受体拮抗剂昂丹司琼不仅被提议用于酒精依赖的标签外治疗，而且也被广泛使用。 HTR3D 用作怀孕期间女性的止吐剂，在该模型的背景下似乎是值得考虑的有趣候选者。迄今为止，HTR3D 基因仅在人类、黑猩猩和狗中得到表征。在这些物种中，HTRD3 基因与 HTR3C 和 HTR3E 存在于一个簇中，并且似乎主要局限于背根神经节 (DRG) 的表达，尽管在其他神经组织（包括额叶）中观察到低水平的表达。叶和下丘脑。 DRG 中的高水平表达很有趣，因为我们筛选的 70% 的动物如果是替代 rhHTR3D 等位基因的携带者，则具有高感觉防御能力。由于我们的初步筛选表明 HTR3D 变异可能在感觉处理中发挥作用，因此我们想要检查感觉敏感性的个体差异是否会根据产前乙醇暴露和基因型而变化。我们发现 rhHTR3D 基因型与产前暴露时间之间存在相互作用。基因型似乎可以预测早期酒精暴露不同的动物的平均感觉强度（对触觉刺激的退缩和无法习惯轻触），但在妊娠晚期暴露的动物中，根据母亲在早期是否饮酒，存在定性交互效应。怀孕。尽管样本很小，但报告 FAE 遗传风险因素的研究很少，而且对 HTR3D 基因知之甚少，这使得我们的发现很有趣。该基因在啮齿类动物中似乎不存在，这说明了在灵长类动物中进行的研究具有巨大的转化价值。人类酒精依赖样本中是否存在 HTR3D 突变，这对于预测昂丹司琼反应可能很重要，这似乎是合理的。在这里，我们发现等位基因剂量效应可用于预测猕猴自愿乙醇自我给药的个体差异。如上所述，HT-3受体拮抗剂已被用于治疗酒精成瘾。实际上，终止密码子变体可以充当该受体的拮抗剂。正如人们所预料的那样，这种功能丧失变异与较低水平的自我给药有关。也许 NIH 恒河猴种群（几十年前维持的）最有价值的特征是，至少有 12 只接受评估的个体是在托儿所中饲养的（PR，同伴饲养）。在这些条件下饲养的猴子通常会变得更加焦虑、冲动或具有攻击性，并且存在社会认知差异以及依恋障碍的特征。当有选择时，他们也会自愿喝更多的酒。由于早期饲养模式，我们有机会研究 G x E 相互作用。至少，从该群体存档的数据集提供了对由于早期环境暴露而表现出行为病理学的个体丰富的群体的行为表型的访问。这可能会提高进行遗传学研究的敏感性，以模拟人类发展精神病理学和酒精问题的轨迹。来自应激动物的脑组织的 RNA 测序可能对于识别功能重要基因内的变异特别有用。通过对早期饲养历史不同的猕猴大脑中存档的样本进行 RNA 测序，我们可以筛选行为相关和压力反应大脑区域中表达的基因的变异。使用 RNA-Seq 在恒河猴中检测到的 SNP 中包括伤害感受肽/孤啡肽 FQ（rhOPRL1，也称为 kappa 3）受体 (rhOPRL1 His243Arg) 中的 nsSNP。与其他阿片受体一样，痛敏肽/孤啡肽 FQ 受体除了与压力和物质使用相关的表型外，还赋予对疼痛的不同敏感性。 rhOPRL1 His243Arg SNP 位于此类受体中高度保守的区域，polyphen 预测该 SNP 具有破坏性（得分为 0.999）。有证据表明，具有重要功能的 OPRL1 变异存在于不同物种之间。在人类中，OPRL1 的表达在自闭症谱系障碍和阿尔茨海默病中发生改变，而在狗中，古老的错义多态性已经固定在某些家养狗品种中（特别是水猎犬，经过选择性培育以承受冷水浸泡），这种表型可能会导致在人类冷加压/疼痛反应的背景下考虑。在实验室啮齿动物中进行的研究表明，通过给予伤害感受素受体拮抗剂，可以抑制因酒精偏好而选择的菌株的乙醇自我给药。这表明该受体的遗传变异可能导致酒精偏好表型。这些研究表明，受体的拮抗作用不仅会阻止条件性位置偏好的获得，还会阻止通过间歇性访问过渡到依赖状态的速率。依赖后的动物中压力和提示诱导的恢复也受到阻碍。由于这些原因，该受体的拮抗作用已被提议作为酒精成瘾的潜在治疗方法。目前还缺乏研究 OPRL1 自然发生的遗传变异与成瘾相关表型和受体拮抗剂的药物遗传学相关的作用的遗传学研究。我们发现 rhOPRL1 的破坏性变异可以预测恒河猴早期酒精摄入量的个体差异。尽管没有交互作用，但将早期经验纳入模型似乎对该样本很重要。可以想象，OPRL1 基因型效应在先前压力负担较高的人群中可能更明显。通过获得口服生物可利用的伤害感受受体拮抗剂（被提议作为 MDD 和酒精使用障碍的潜在治疗方法），旨在检查 OPRL1 的药物遗传学效应的研究也可能在人类中进行。如上所述，人类和猕猴体内都存在功能性 Mu-阿片受体变异体。鉴于这些基因的变异在预测对社会排斥和共情反应的痛苦敏感性差异方面存在联系，这两个基因的变异也可能与已知与 AUD 共病的人格障碍风险有关。