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.

随着下一代测序进行和完善基因组测序的能力的扩展，我们开始在非人类灵长类动物中揭示遗传和环境因素，这有助于在包括成瘾在内的各种人类神经发育和精神疾病中观察到的个人性状变化。使用存档的数据集并依靠正在进行的协作，我们一直在从合理尺寸的数据集中得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。我们假定会发生人类酒精饮酒障碍的潜在途径，其中一些将在下面进行讨论。我们对在子宫内暴露于酒精的猕猴进行了整个外显子组测序（WES）。使用具有极端FAE表型的个体（高/搅动与平静/柔和的气质，高与低分离引起的ACTH响应，最佳与最差的试验，对于延迟样本的标准，高/防御性与低/不反应的标准，对于感觉处理的最佳试验），我们发现了定位型的NEURON和NEURON的功能，该neuron and neuron neuron and neuron neuron neuron neuron neuron neuron neuron，对于5-羟色胺HT-3受体（RHHTR3D）。鉴于5-羟色胺系统在焦虑中的作用，除了HT-3受体拮抗剂ondansetron不仅提出了用于酒精依赖的标签治疗，而且在怀孕期间对妇女的抗态度，HTR3D似乎是一种有趣的候选者，也被用作妇女的抗态度。到目前为止，HTR3D基因仅在人类，黑猩猩和狗中被表征。在这些物种中的每一个中，HTRD3基因与Htr3c和Htr3E的簇存在，并且似乎在其表达中大部分限于背根神经节（DRG），尽管在包括额叶叶和下丘脑在内的其他神经元组织中观察到的表达水平较低。 DRG中高水平的表达很有趣，因为如果我们筛选的动物是替代性RHHTR3D等位基因的载体，我们筛选的70％的动物是高感觉防御的。正如我们的初始屏幕表明HTR3D变异可能在感觉过程中起作用的可能性，我们想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基因型和产前暴露的时间之间的相互作用。基因型似乎可以预测平均感觉幅度（从触觉刺激和未能习惯于轻触摸的动物中撤出，这些动物因早期酒精暴露而发散，但是在晚期妊娠暴露的动物中，根据母亲在怀孕早期饮酒中是否饮酒会产生定性的互动效应。尽管样本很小，但有很少的研究报告了FAE的遗传危险因素，而对HTR3D基因的了解很少，这使我们的发现令人着迷。该基因在啮齿动物中似乎不存在，这说明了灵长类动物中进行的研究的巨大翻译价值。在人类酒精依赖性样品中是否有HTR3D突变在预测Ondansetron反应中可能很重要。在这里，我们发现在预测猕猴中自愿性乙醇自我给药的个体差异时，存在等位基因剂量效应。如上所述，HT-3受体拮抗作用已用于治疗酒精成瘾。终止密码子变体实际上可以作为该受体的拮抗剂。正如人们可能期望的那样，这种功能变异的损失与较低的自我给药水平有关。 NIH恒河总数最有价值的特征（几十年前维持）也许是至少有12名评估的人是在托儿所中饲养的（PR，Peer饲养）。在这些条件下饲养的猴子通常会变得更加焦虑，冲动或侵略性，并且具有社会认知差异以及依恋障碍的特征。当选择时，他们还会自愿食用更多的酒精。由于早期饲养范式，我们提供了检查G X E相互作用的机会。至少，从该殖民地存档的数据集为由于早期环境暴露而富含行为病理的人群提供了行为表型的访问权限。这可能会增加对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。来自压力暴露动物的脑组织的RNA序列可能对于鉴定功能上重要的基因内的变异特别有用。通过对从分歧为早期饲养历史的个体中猕猴的样品进行测序的RNA，我们可以筛选在行为相关和应激响应性的大脑区域中表达的基因内的变化。在恒河猕猴中使用RNA-Seq检测到的SNP中，是NSSNP，是NSSNP/orphanin FQ（Rhoprl1，也称为Kappa 3）受体（Rhoprl1 His243arg）。与其他阿片类药物受体一样，除了与胁迫和药物使用有关的表型外，诺耐匹毒素/孤儿FQ受体除了表型外，还对疼痛的差异敏感。 RHOPRL1 HIS243ARG SNP位于这类受体高度保守的区域中，而多芯片预测该SNP具有损害（得分为0.999）。有证据表明，功能上重要的OPRL1变化发生在各种物种之间。在人类中，OPRL1的表达在自闭症谱系障碍和阿尔茨海默氏病中发生了改变，在狗中，古老的错过多态性的多态性已经固定在某些家养狗品种中（尤其是水犬，有选择性地培养以承受在冷水中沉浸在冷水中），这是一种在人类冷persor/painer paintor/painer painter/painer paintor/pain的表型。在实验室啮齿动物中进行的研究表明，选择用于酒精偏爱的菌株中的乙醇自我给药可以抑制Nocceptin受体拮抗剂。这表明该受体的遗传变异可能有助于酒精偏好表型。这些研究表明，受体的拮抗作用不仅阻止了条件位置偏好的获取，而且还阻止了过渡到依赖状态的速率，并具有间歇性通道。在依赖后动物中，应力和提示引起的恢复也被阻塞。由于这些原因，已经提出了该受体的拮抗作用，以作为对酒精成瘾的潜在治疗方法。缺乏遗传研究，研究了与成瘾相关的表型和受体拮抗剂的药物遗传学有关的OPRL1天然遗传变异的作用。我们表明，Rhoprl1处的破坏变化预测了恒河猕猴早期获得酒精摄入量的个体差异。尽管没有相互作用，但在模型中包含早期经验对于该样本似乎很重要。可以想象，在较高的人群中，OPRL1基因型效应可能更为明显。通过获得口服生物学上的伤害受体拮抗剂，提议作为MDD和酒精使用障碍的潜在治疗方法，旨在检查OPRL1药物遗传学作用的研究也可能在人类中也可能存在。如上所述，人类和猕猴都有功能性的MU阿片受体变体。鉴于这些基因的变异之间的联系在预测对社会拒绝和移情反应疼痛敏感性差异方面的差异，这两个基因的变异也可能与已知的人格障碍的风险有关。