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.

项目总结/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。导致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，通过确定转录如何解决这一差距， 记忆相关基因的调节在男性和女性中不同，以支持恐惧记忆。最近我们 研究小组证明，雄性和雌性大鼠在前额叶内的信号传导机制存在差异， 我们的初步研究结果表明，在恐惧记忆的形成过程中， 前额叶基因的表观遗传修饰。目前，情景记忆的转录调控， 人们对认知系统知之甚少，性对这一动态过程的影响也几乎一无所知。这 这对开发有效治疗诸如PTSD的疾病是一个重大挑战。客观 这项研究的一个重要目的是确定DNA羟甲基化酶泰特如何在功能上调节恐惧记忆 形成和识别厌恶体验的性别特异性甲基化特征。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， 与大脑中恐惧记忆的形成有关然而，是否存在性别差异的参与， 恐惧记忆形成过程中的DNA 5-hmC机制仍然不明确。在我们的初步研究中， 在大鼠前额叶皮层中，DNA羟甲基化酶Tet 2在雌性大鼠中增加，而Tet 1在雌性大鼠中减少。 男性学习后的痕迹恐惧条件反射范例。核心假设是TET介导的 前额叶皮层性别特异性基因靶点的DNA 5-羟甲基化促进恐惧的形成 记忆该方法使用无偏全基因组分析（NIMH策略1.2，研究优先级B）， 与病毒介导的转录控制组合，以测试泰特活性与 NIMH策略1.1，研究优先级D; NIMH策略1.2，研究优先级B。目标1将确定 恐惧后TET 1和TET 2介导的前额叶皮层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（ChIP）和羟甲基化 DNA免疫沉淀（hmeDIP）测序方法。Aim 2将使用最先进的CRISPR-dCas 9 技术来操纵Tet 1和Tet 2在男性和女性的前额叶皮层的表达， 微量恐惧条件反射，这将决定皮质TET 1和TET 2在性别依赖性功能中的作用， 恐惧记忆的形成这项拟议中的研究意义重大，因为它有望提供一个（epi）基因组 性二态大脑区域（内侧前额叶皮质）的记忆形成地图，这是进行以下活动所需的 了解，甚至确定新的遗传生物标志物的性别偏见的情绪和认知缺陷， 精神病

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.