Genome-wide analysis of sex differences in cortical DNA hydroxymethylation during fear memory formation

恐惧记忆形成过程中皮质 DNA 羟甲基化的性别差异的全基因组分析

• 批准号: 10041818
• 负责人: MARIEKE R GILMARTIN
• 金额: $ 42.58万
• 依托单位: VIRGINIA POLYTECHNIC INST AND ST UNIV
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-20 至 2023-07-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10041818
• 关键词: Address; Adult; Affect; Alzheimer' s Disease; Animals; Anxiety; Behavioral; Biological; Biological Markers; Brain; Brain Diseases; Brain region; Cells; ChIP-seq; Clustered Regularly Interspaced Short Palindromic Repeats; Cognitive deficits; DNA; Data; Development; Disease; Emotional; Enzymes; Epigenetic Process; Episodic memory; Extinction (Psychology); Female; Fright; Gene Expression; Gene Targeting; Generations; Genes; Genetic Markers; Genetic Transcription; Genomics; Immunoprecipitation; Lead; Learning; Link; Maps; Medial; Mediating; Memory; Mental disorders; Methods; Methylation; Modification; Molecular; National Institute of Mental Health; Neurobiology; Outcome; Population; Post-Traumatic Stress Disorders; Pre-Clinical Model; Predisposition; Prefrontal Cortex; Process; Public Health; Rattus; Research; Research Priority; Resistance; Risk Factors; Role; Sex Bias; Sex Differences; Signal Transduction; Synaptic plasticity; System; Technology; Testing; Therapeutic; Training; Transcription Process; Transcriptional Regulation; Trauma; Viral; Woman; Work; chromatin immunoprecipitation; classical conditioning; cognitive system; conditioned fear; demethylation; differential expression; effective therapy; epigenetic regulation; experience; experimental study; fear memory; genetic analysis; genome wide association study; genome-wide analysis; innovation; insight; male; memory consolidation; men; methylome; methylomics; next generation; novel; pre-clinical; resilience; sex; sexual dimorphism; transcriptomics; traumatic event; whole genome

Project Summary/Abstract Post-traumatic stress disorder (PTSD) affects nearly 8% of the population and is more prevalent in women than men. The neurobiological factors that contribute to this sex bias are largely unknown. This project addresses this gap, in accordance with NIMH Strategic Objective 1, by determining how transcriptional regulation of memory-related genes differs in males and females in support of fear memory. Recently our group demonstrated that male and female rats differentially engage signaling mechanisms within the prefrontal cortex (PFC) during the formation of a fear memory, and our preliminary findings point to sex differences in epigenetic modification of prefrontal genes. Currently, the transcriptional regulation of episodic memory within cognitive systems is poorly understood, and the impact of sex on this dynamic process is all but unknown. This represents a significant challenge to developing effective treatment for disorders such as PTSD. The objective of this proposal is to determine how DNA hydroxymethylase TET enzymes functionally regulate fear memory formation and to identify sex-specific methylomic signatures of aversive experience. DNA 5- hydroxymethylation (5-hmC), a major regulator of active (increased) transcription in cells, has been recently implicated in fear memory formation in the brain. However, whether sex differences exist in the engagement of DNA 5-hmC mechanisms during fear memory formation remain equivocal. In our preliminary studies we found that in the rat prefrontal cortex the DNA hydroxymethylase Tet2 increased in females while Tet1 decreased in males following learning in a trace fear conditioning paradigm. The central hypothesis is that TET-mediated DNA 5-hydroxymethylation of sex-specific gene targets in the prefrontal cortex facilitates the formation of fear memory. The approach uses unbiased genome-wide analysis (NIMH Strategy 1.2, Research Priority B) in combination with viral-mediated transcriptional control to test the functional link between TET activity and memory (NIMH Strategy 1.1, Research Priority D; NIMH Strategy 1.2, Research Priority B). Aim 1 will identify the sex-specific gene targets of TET1 and TET2-mediated DNA 5-hmC in the prefrontal cortex following fear learning, using next generation whole genome chromatin immunoprecipitation (ChIP) and hydroxymethylated DNA immunoprecipitation (hmeDIP) sequencing methods. Aim 2 will use cutting-edge CRISPR-dCas9 technology to manipulate the expression of Tet1 and Tet2 in the prefrontal cortex of males and females during trace fear conditioning, which will determine the sex-dependent functional role of cortical TET1 and TET2 in fear memory formation. The proposed research is significant because it is expected to provide a (epi)genomic map of memory formation in a sexually dimorphic brain region, the medial prefrontal cortex, that is needed for understanding and even identifying new genetic biomarkers of sex-biased emotional and cognitive deficits in psychiatric illness.

项目摘要/摘要 创伤后应激障碍(PTSD)影响了近8%的人口，在女性中更为普遍 而不是男人。造成这种性别偏见的神经生物学因素在很大程度上是未知的。这个项目 根据NIMH战略目标1，通过确定转录如何 男性和女性对支持恐惧记忆的记忆相关基因的调控是不同的。最近我们的 研究小组证明，雄性和雌性大鼠在前额叶内参与不同的信号机制 大脑皮层(PFC)形成恐惧记忆，我们的初步发现指出性别差异在 前额叶基因的表观遗传修饰。目前，情景记忆在脑内的转录调控 人们对认知系统知之甚少，性行为对这一动态过程的影响也几乎是未知的。这 这是为创伤后应激障碍等疾病开发有效治疗方法的重大挑战。目标是 这项建议的目的是确定DNA羟甲基酶Tet酶如何调节恐惧记忆的功能 形成和识别厌恶体验的性别特定的甲基组特征。DNA 5- 羟甲基化(5-HMC)是细胞中活跃(增加)转录的主要调节因子，最近 与大脑中恐惧记忆的形成有关。然而，在订婚中是否存在性别差异 DNA 5-HMC在恐惧记忆形成过程中的作用机制仍不明确。在我们的初步研究中我们发现 雌性大鼠前额叶皮质DNA羟甲基酶TET2增加，Tet1减少。 在痕迹恐惧条件反射范式中，男性跟随学习。中心假设是Tet介导的 前额叶皮质性别特异性基因靶点DNA 5-羟甲基化促进恐惧的形成 记忆。该方法使用了无偏见的全基因组分析(NIMH策略1.2，研究优先B) 结合病毒介导的转录控制来测试Tet活性和Tet活性之间的功能联系 存储器(NIMH策略1.1，研究优先D；NIMH策略1.2，研究优先B)。目标1将确定 恐惧后前额叶皮质TET1和TET2介导的DNA 5-HMC的性别特异性基因靶点 学习，使用下一代全基因组染色质免疫沉淀(芯片)和羟甲基化 DNA免疫沉淀(HmeDIP)测序法。AIM 2将使用尖端的CRISPR-dCas9 调控Tet1和TET2在成年男性和女性前额叶皮质表达的技术 微量恐惧条件反射，这将决定大脑皮质TET1和TET2的性别依赖性功能作用 恐惧记忆的形成。这项拟议的研究意义重大，因为它有望提供(EPI)基因组 大脑内侧前额叶皮质这一性别二态大脑区域的记忆形成图，这是 了解甚至识别性别偏见的情绪和认知缺陷的新的遗传生物标志物 精神疾病。

项目成果

Epigenetic Mechanisms in Memory and Cognitive Decline Associated with Aging and Alzheimer's Disease.

• DOI: 10.3390/ijms222212280
• 发表时间: 2021-11-13
• 期刊: International journal of molecular sciences
• 影响因子: 5.6
• 作者: Maity S;Farrell K;Navabpour S;Narayanan SN;Jarome TJ
• 通讯作者: Jarome TJ

Females, but not males, require protein degradation in the hippocampus for contextual fear memory formation.

• DOI: 10.1101/lm.053429.121
• 发表时间: 2021-08
• 期刊: Learning & memory (Cold Spring Harbor, N.Y.)
• 作者: Martin K;Musaus M;Navabpour S;Gustin A;Ray WK;Helm RF;Jarome TJ
• 通讯作者: Jarome TJ

Sex-differences in proteasome-dependent K48-polyubiquitin signaling in the amygdala are developmentally regulated in rats.

• DOI: 10.1186/s13293-023-00566-z
• 发表时间: 2023-11-10
• 期刊: BIOLOGY OF SEX DIFFERENCES
• 影响因子: 7.9
• 作者: Farrell, Kayla;Auerbach, Aubrey;Liu, Catherine;Martin, Kiley;Pareno, Myasia;Ray, W. Keith;Helm, Richard F.;Biase, Fernando;Jarome, Timothy J.
• 通讯作者: Jarome, Timothy J.

Males and females differ in the regulation and engagement of, but not requirement for, protein degradation in the amygdala during fear memory formation.

• DOI: 10.1016/j.nlm.2021.107404
• 发表时间: 2021-04
• 期刊: Neurobiology of learning and memory
• 影响因子: 2.7
• 作者: Devulapalli R;Jones N;Farrell K;Musaus M;Kugler H;McFadden T;Orsi SA;Martin K;Nelsen J;Navabpour S;O'Donnell M;McCoig E;Jarome TJ
• 通讯作者: Jarome TJ

Protein SUMOylation is a sex-specific regulator of fear memory formation in the amygdala.

• DOI: 10.1016/j.bbr.2022.113928
• 发表时间: 2022-07-26
• 期刊: BEHAVIOURAL BRAIN RESEARCH
• 影响因子: 2.7
• 作者: Gustin, Aspen;Navabpour, Shaghayegh;Farrell, Kayla;Martin, Kiley;DuVall, Jessica;Ray, W. Keith;Helm, Richard F.;Jarome, Timothy J.
• 通讯作者: Jarome, Timothy J.