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.

项目摘要/摘要 精神分裂症（SZ）和严重抑郁症等重大精神疾病涉及社会异常 行为和发作通常发生在青春期后。 SZ的遗传危险因素经常是杂合子 在受影响的个体中，临床表型可能是高度可变的。但是，贡献的因素 对于表型差异和这些疾病发作的时机的理解很少。基因组 印迹是一种表观遗传机制，可引起母亲或父亲等位基因的优先表达 一些基因。烙印会导致产卵的效应，从而影响遗传的表型效应 突变。规范的不张纸涉及完全沉默一个父母的等位基因，并影响少量 基因。但是，在最近的出版物中，作者的实验室发现了鼠标中的数百个基因 暴露了母子或父亲等位基因表达偏见。这种现象称为非规范印迹。 与肝脏或肌肉相比，非规范的印迹在大脑中高度富集。使用小说等位基因 - 特定的原位杂交方法，作者发现非规范印迹基因暴露了单相的基因 在大脑中细胞的亚群中的表达，表明在 脑。目前，尚不知道非规范印记是一种细胞类型的特定形式的印迹 脑。此外，该领域不知道印在特定细胞类型中的基因的身份 大脑，非规范印迹是否在青春期可能会发生变化，或者社会行为是否受到影响。 作者发现酪氨酸羟化酶（Th）和DOPA脱羧酶（DDC）是非规范印迹基因 在大脑的特定区域中具有母体等位基因偏见。 TH和DDC合成单胺 神经递质在社会行为和精神疾病中具有重要作用。在这里，作者将测试 单胺能电路中非规范印迹效应是一种高度细胞类型的假设 可能会因青春期发展而影响社会的特定形式的不贴图 在精神疾病中扮演角色的行为。 AIM 1（K99）将确定非规范印迹是否是细胞 单胺系统中的印迹的特定形式，并揭示了基因的身份 印在大脑中单胺能神经元的分子定义亚型中。这些研究涉及 产生母校和父亲等位基因特异性记者小鼠，以在DDC的细胞水平上解决印迹 以及来自不同不同的单胺能神经元中细胞水平上印迹的新方法 使用单细胞RNASEQ的大脑区域。 AIM 2（R00）将确定是否在 在青春期前后，在青春期发生变化时，前后核（ARN）变化。 使用我们实验室中开发的RNASEQ方法的青春期。 POA和ARN在青春期和 社会行为和初步数据表明，在这些地区，非规范的印迹可能会改变 对马信号的响应。在AIM 3（R00）中，TH和DDC单和复合突变小鼠将用于 确定是否在相同途径功能中影响基因的非规范无打印效应是否影响 与放大器父母对后代社会行为的影响协同。受影响的社会行为 在精神疾病中将进行测试，包括积极，社会偏好，社会认可，社会奖励和 生殖行为。这些实验将为鼠标基因组工程提供必不可少的新培训 和基因组学（K99），并建立了专注于独立研究计划（R00）的基础 非规范印迹基因和昆虫信号在社会行为和心理健康中的作用。 该提案的K99部分将为主要研究者（PI）提供最新的研究培训 ART CRISPR-CAS9介导的鼠标基因组工程，生物信息学和基因组的编程 分析和单细胞转录组学。职业发展到独立性将由定期教师组成 导师，同事和教职委员会会议，以确保正在研究和职业基准 及时满足。 Mentalship团队将有助于申请和谈判 教师职位的过程，准备独立的研究建议和工作谈判以及提出 在离开大学之前 犹他州。工作谈判和R01提案也将提交给“教师智囊团”，以获得至关重要的 来自整个部门的反馈。主要调查员还将通过 介绍正在进行的研究与部门和相关期刊俱乐部进行对话，参加了科学 PI将有机会进行口头演讲并参加赠款技巧的会议 犹他大学提供的课程。结合独立研究计划的基础， 职业发展活动将确保PI准备进入独立教师 在一所主要研究的大学中的职位，并成为印迹，社会领域的领导者 行为和心理健康。