Genomic imprinting in circuits for social behavior

社会行为回路中的基因组印记

• 批准号: 9224784
• 负责人: Paul Jeffrey Bonthuis
• 金额: $ 9.2万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-02-01 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9224784
• 关键词: ARHGEF5 gene; Adult; Affect; Aggressive behavior; Alleles; Alpha Cell; Applications Grants; Behavior; Behavioral Assay; Benchmarking; Bioinformatics; Brain; Brain region; CRISPR/Cas technology; Cells; Communication; DOPA decarboxylase; Data; Development; Ensure; Enzymes; Epigenetic Process; Exhibits; Faculty; Feedback; Female; Foundations; Genes; Genetic; Genome engineering; Genomic Imprinting; Genomic approach; Genomics; Heterozygote; Hormonal; Hormones; Hybrids; Hypothalamic structure; In Situ Hybridization; Individual; Inherited; Journals; Label; Lead; Link; Liver; Major Depressive Disorder; Mediating; Mediation; Mental Depression; Mental Health; Mental disorders; Mentors; Mentorship; Methods; Molecular; Mus; Muscle; Mutant Strains Mice; Mutation; Neurodevelopmental Disorder; Neurons; Neurotransmitters; Occupations; Onset of illness; Oral; Parents; Pathway interactions; Phenotype; Play; Population; Positioning Attribute; Preoptic Areas; Preparation; Process; Puberty; Public Health; Publications; Reporter; Reproductive Behavior; Research; Research Personnel; Research Proposals; Research Training; Rewards; Role; Schizophrenia; Signal Transduction; Social Behavior; Structure of nucleus infundibularis hypothalami; System; Testing; Time; Training; Training Programs; Transgenic Organisms; Tyrosine 3-Monooxygenase; Universities; Utah; career; career development; cell type; clinical phenotype; experimental study; faculty mentor; genetic approach; genetic risk factor; imprint; improved; innovation; male; meetings; mental development; monoamine; mouse genome; mutant; novel; offspring; parental influence; preference; programs; response; skills; social; transcriptome; transcriptome sequencing; transcriptomics

Project Summary/Abstract Major psychiatric disorders, such as schizophrenia (SZ) and major depression, involve abnormal social behaviors and onset typically occurs after puberty. The genetic risk factors for SZ are frequently heterozygous in affected individuals and the clinical phenotypes can be highly variable. However, the factors that contribute to phenotypic variance and the timing of the onset of these disorders are poorly understood. Genomic imprinting is an epigenetic mechanism that causes preferential expression of the maternal or paternal allele for some genes. Imprinting causes parent-of-origin effects that influence the phenotypic effects of inherited mutations. Canonical imprinting involves complete silencing of one parent's allele and impacts a small number of genes. However, in a recent publication, the author's lab discovered that hundreds of genes in the mouse that exhibit a maternal or paternal allele expression bias. This phenomenon is called noncanonical imprinting. Noncanonical imprinting is highly enriched in the brain compared to the liver or muscle. Using a novel allele- specific in situ hybridization approach, the author found that noncanonical imprinted genes exhibit monoallelic expression in subpopulations of cells in the brain, suggesting a highly cell-type specific form of imprinting in the brain. Currently, it is not known whether noncanonical imprinting is a cell-type specific form of imprinting in the brain. Further, the field does not know the identity of the genes that are imprinted in specific cell-types of the brain, whether noncanonical imprinting can change at puberty or whether social behaviors are impacted. The author found that tyrosine hydroxylase (Th) and dopa decarboxylase (Ddc) are noncanonical imprinted genes with a maternal allele bias in specific regions of the brain. TH and DDC synthesize monoamine neurotransmitters, which have important roles in social behavior and mental illness. Here, the author will test the hypothesis that noncanonical imprinting effects in monoaminergic circuits are a highly cell-type specific form of imprinting that can change in response to pubertal development and impact social behaviors with roles in mental illness. Aim 1 (K99) will determine whether noncanonical imprinting is a cell- type specific form of imprinting in the monoamine system and uncover the identity of the genes that are imprinted in molecularly-defined subtypes of monoaminergic neurons in the brain. These studies involve generating maternal and paternal allele-specific reporter mice to resolve imprinting at the cellular level for Ddc and new methods to profile imprinting at the cellular level in isolated monoaminergic neurons from different brain regions using single-cell RNASeq. Aim 2 (R00) will determine whether noncanonical imprinting in the preoptic area (POA) and arcuate nucleus (ARN) changes at puberty by profiling imprinting before and after puberty using RNASeq methods developed in our lab. The POA and ARN play important roles in puberty and social behaviors and preliminary data shows that noncanonical imprinting can change in these regions in response to hormonal signaling. In Aim 3 (R00), Th and Ddc single and compound mutant mice will be used to determine whether noncanonical imprinting effects impacting genes in the same pathway function synergistically to amplify parental influences on offspring social behaviors. Social behaviors that are impacted in mental illness will be tested, including aggression, social preference, social recognition, social reward and reproductive behaviors. These experiments will provide essential new training in mouse genome engineering and genomics (K99) and establish the foundations for an independent research program (R00) focused on the roles of noncanonical imprinted genes and hormone signaling in social behaviors and mental health. The K99 portion of this proposal will provide the principle investigator (PI) with research training in state-of-the art CRISPR-Cas9 mediated mouse genome engineering, programming for bioinformatics and genomic analysis, and single cell transcriptomics. Career development to independence will consist of regular faculty mentor, co-mentor, and faculty committee meetings to ensure that research and career benchmarks are being met in a timely and appropriate fashion. The mentorship team will aid in the application and negotiation process for faculty positions, the preparation of an independent research proposal and job talks, and putting together the Aims and Research Strategy for the PI's first R01 grant application before leaving the University of Utah. The job talks and R01 proposals will also be presented to the “Faculty Think Tank” to get critical feedback from the whole department. The principle investigator will also improve communication skills by presenting research in progress talks to the department and relevant journal clubs, attending scientific meetings where the PI will have an opportunity to give oral presentations, and attending a grantsmanship course offered at the University of Utah. Combined with the foundation of the independent research program, the career development activities will ensure that the PI is well prepared to enter an independent faculty position at a major research oriented university and emerge as a leader in the fields of imprinting, social behavior and mental health.

项目概要/摘要 主要精神疾病，例如精神分裂症（SZ）和重度抑郁症，涉及异常的社交 SZ 的行为和发病通常发生在青春期之后。 SZ 的遗传危险因素通常是杂合的。 在受影响的个体中，临床表型可能存在很大差异，但影响因素却多种多样。 对表型变异和这些疾病的发病时间的基因组了解甚少。 印记是一种表观遗传机制，导致母本或父本等位基因优先表达 一些基因的印记会导致影响遗传表型效应的亲本效应。 典型印记涉及父母之一的等位基因的完全沉默并影响一小部分。 然而，在最近的一篇出版物中，作者的实验室发现了小鼠体内的数百个基因。 表现出母系或父系等位基因表达偏差的现象称为非规范印记。 与肝脏或肌肉相比，非规范印记在大脑中高度丰富。 通过特定的原位杂交方法，作者发现非典型印记基因表现出单等位基因 在大脑细胞亚群中表达，表明大脑中存在高度细胞类型特异性的印记形式 目前，尚不清楚非规范印记是否是大脑中细胞类型特定的印记形式。 此外，该领域并不知道印记在大脑特定细胞类型中的基因的身份。 大脑的非规范印记是否会在青春期发生变化，或者社会行为是否会受到影响。 作者发现酪氨酸羟化酶（Th）和多巴脱羧酶（Ddc）是非经典印记基因 TH 和 DDC 合成单胺具有母体等位基因偏差。 在这里，作者将测试在社会行为和精神疾病中发挥重要作用的神经递质。 单胺能回路中的非典型印记效应是高度细胞类型的假设 特定形式的印记可以随着青春期的发展而改变并影响社会 目标 1 (K99) 将确定非规范印记是否是一种细胞- 在单胺系统中键入特定形式的印记并揭示其基因的身份 这些研究涉及大脑中单胺能神经元的分子定义亚型。 生成母本和父本等位基因特异性报告小鼠，以解析 Ddc 的细胞水平印记 以及在细胞水平上分析来自不同来源的分离的单胺能神经元的印记的新方法 使用单细胞 RNASeq 2 (R00) 的大脑区域将确定是否存在非规范印记。 通过分析印记前后的情况观察青春期视前区（POA）和弓状核（ARN）的变化 使用我们实验室开发的 RNASeq 方法的青春期 POA 和 ARN 在青春期和青春期中发挥着重要作用。 社会行为和初步数据表明，这些区域的非规范印记可能会发生变化 在 Aim 3 (R00) 中，Th 和 Ddc 单一和复合突变小鼠将用于 确定非典型印记效应是否影响同一通路功能中的基因 协同作用，放大父母对后代社会行为的影响。 在精神疾病中将受到测试，包括攻击性、社会偏好、社会认可、社会奖励和 这些实验将为小鼠基因组工程提供重要的新训练。 和基因组学（K99），并为专注于以下方面的独立研究计划（R00）奠定基础： 非典型印记基因和激素信号在社会行为和心理健康中的作用。 该提案的 K99 部分将为首席研究员 (PI) 提供最新研究培训 CRISPR-Cas9介导的小鼠基因组工程、生物信息学和基因组学编程 分析和单细胞转录组学的职业发展将由常规教师组成。 导师、共同导师和教师委员会会议，以确保研究和职业基准得到落实 及时、适当的方式会面，指导团队将协助申请和谈判。 教师职位的流程，独立研究计划和工作演讲的准备，以及 在离开大学之前，将 PI 的第一个 R01 资助申请的目标和研究策略结合在一起 犹他州的工作会谈和 R01 提案也将提交给“教职智库”以获得批评。 整个部门的反馈也将提高沟通技巧。 向系和相关期刊俱乐部介绍正在进行的研究讲座，参加科学会议 PI 将有机会进行口头演讲并参加资助的会议 结合犹他大学提供的独立研究项目的基础， 职业发展活动将确保 PI 为进入独立学院做好充分准备 在一所主要研究型大学中占据一席之地，并成为印记、社会领域的领导者 行为和心理健康。