Studies of histone serotonylation and its role in transcriptional and neural plasticity

组蛋白血清酰化及其在转录和神经可塑性中的作用的研究

• 批准号: 10016853
• 负责人: Lorna Farrelly
• 金额: $ 9.05万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-11 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10016853
• 关键词: Address; Adult; Affect; Antibodies; Antidepressive Agents; Anxiety; Award; Behavioral; Binding; Binding Proteins; Biochemical; Biochemistry; Bioinformatics; Biological; Biological Assay; Brain; Cell Nucleus; Cell membrane; ChIP-seq; Chromatin; Chronic; Chronic stress; Clinical; Complex; Coupled; Cytoplasmic Protein; Data; Deacetylase; Development; Disease; Dominant-Negative Mutation; Dopamine; Dorsal; Embryonic Development; Environment; Environmental Exposure; Enzymes; Epigenetic Process; Euchromatin; Exons; Foundations; Functional disorder; Gastrointestinal Motility; Gene Expression; Genes; Genetic Transcription; Glutamine; Histone H3; Histones; Human; Immunoprecipitation; In Vitro; Individual; Investigation; Laboratories; Literature; Mental disorders; Mentors; Mentorship; Methodology; Modeling; Modification; Molecular; Moods; Mus; Nature; Neurodegenerative Disorders; Neurodevelopmental Disorder; Neuroglia; Neuronal Plasticity; Neurons; New York; Norepinephrine; Nuclear; Nuclear Proteins; Nucleosomes; Output; Pattern; Peptides; Peripheral; Phase; Phenotype; Physiologic Thermoregulation; Physiological; Play; Process; Property; Proteins; Regulation; Research; Rodent; Role; Selective Serotonin Reuptake Inhibitor; Serotonin; Sex Behavior; Signal Transduction; Stress; Syndrome; TAF1 gene; Tissues; Training; Validation; Western Blotting; Work; adeno-associated viral vector; behavioral plasticity; brain research; career; cell type; circadian; combinatorial; covalent bond; dorsal raphe nucleus; epigenomics; genetic manipulation; genome-wide; genome-wide analysis; in vivo; induced pluripotent stem cell; insight; liquid chromatography mass spectrometry; mature animal; medical schools; mutant; nervous system disorder; neurodevelopment; neuron development; neurotransmission; novel; overexpression; permissiveness; post-doctoral training; postnatal; postnatal development; programs; social defeat; transamination; transcriptome sequencing; transglutaminase 2

Project Summary Alterations in serotonin (5HT) dynamics are observed in diverse clinical disorders including, but not limited to, stress and anxiety related syndromes. Interestingly, recent investigations have demonstrated an additional presence of ‘reserve’ pools of extravesicular 5HT in dorsal raphe (DRN) serotonergic neurons (both cytoplasmic and nuclear) however it has remained unclear whether it might play additional roles in the nucleus. A growing literature now demonstrates 5HT’s ability to form covalent bonds with certain cytoplasmic and membrane bound proteins via transamination by the tissue Transglutaminase 2 (TGM2) enzyme, a modification that has been suggested to alter the signaling properties of monoaminylated substrates. We therefore initially hypothesized that nuclear proteins in brain may similarly be modified to control distinct aspects of their function. Work from our group (Farrelly et al., Nature, 2018 – re-review) has shown that 5HT can form covalent bonds with histone proteins (specifically histone H3; H3K4me3Q5ser)–a process known as serotonylation–which is critical for transcriptional permissiveness and early neuronal development. Thus, my plan during this K99/R00 proposal is to further explore the hypothesis that novel histone serotonylation may play critical roles in guiding normal patterns of transcriptional and cellular plasticity in brain during postnatal development and into adulthood, processes that may be disrupted in pathophysiological states associated with serotonergic dysfunction (i.e., chronic stress). Under the mentorship of Drs. Ian Maze and Li Shen at the Icahn School of Medicine at Mount Sinai New York, I will address this hypothesis in three distinct aims using a variety of approaches. During the K99 period (Aim 1) I plan to directly examine associations between H3K4me3Q5ser enrichment and transcriptional (dys)plasticity in developing and adult mouse brain (neurons vs. glia). This will also be investigated using a model of chronic social defeat stress (+/- antidepressants) as a mechanism to perturb serotonin signaling and thus further define a biological role for this modification in vivo. I will then assess (Aim 2) the contributions of H3 serotonylation to adulthood neural plasticity through genetic manipulations that alter levels of the mark, followed by extensive behavioral and transcriptional phenotyping. In my independent research program in Aim3 (R00), I will set out to further dissect distinct roles for H3Q5ser, as my preliminary data indicate that the serotonyl modification can exist in isolation of K4me3 and interacts uniquely in comparison to H3K4me3Q5ser with putative binding complexes (e.g., NuRD). This proposal promises to establish important patterns of transcriptional plasticity associated with histone serotonylation in brain. Moreover, the additional training afforded by this award and the scientific environment will ideally prepare me to launch an independent research program evaluating at multiple levels (epigenomic, cellular/behavioral) as to how histone serotonylation contributes to normal and abnormal states of plasticity.

项目摘要 在多种临床病症中观察到5-羟色胺（5 HT）动力学的改变，包括但不限于， 压力和焦虑相关综合征。有趣的是，最近的调查表明， 中缝背核（DRN）中存在囊外5 HT的“储备”池， 细胞质和细胞核），但仍不清楚它是否可能在细胞核中发挥其他作用。 越来越多的文献证明，5-HT能够与某些细胞质形成共价键， 通过组织转氨酶2（TGM 2）酶的转氨作用， 已经提出改变单胺化底物的信号传导性质的修饰。我们 因此，最初假设大脑中的核蛋白可能类似地被修饰以控制不同的细胞， 其功能的各个方面。我们小组的工作（Farrelly等人，Nature，2018 - re-review）已经表明，5 HT 可以与组蛋白（特别是组蛋白H3; H3 K4 me 3Q 5ser）形成共价键-这一过程被称为 去甲肾上腺素化-这对转录允许性和早期神经元发育至关重要。所以我的 在K99/R 00提案中的一个计划是进一步探索新的组蛋白乙酰化可能 在指导出生后大脑中转录和细胞可塑性的正常模式中起关键作用 发育和进入成年期，这些过程可能在与以下疾病相关的病理生理状态中被破坏： 肾上腺素能功能障碍（即，慢性压力）。在伊坎的伊恩迷宫和李申博士的指导下， 纽约西奈山医学院，我将在三个不同的目标，使用各种解决这一假设 的方法。在K99期间（目标1），我计划直接检查H3 K4 me 3Q 5ser之间的关联 在发育和成年小鼠脑中的富集和转录（dys）可塑性（神经元对神经胶质）。这将 还可以使用慢性社会失败压力（+/-抗抑郁药）模型作为机制进行研究， 干扰5-羟色胺信号传导，从而进一步确定这种修饰在体内的生物学作用。然后我将 评估（目的2）H3羟丙酰化对成年神经可塑性的贡献，通过遗传学方法， 操作，改变水平的标志，其次是广泛的行为和转录表型。在 我在Aim 3（R 00）的独立研究计划中，我将着手进一步剖析H3 Q5 ser的不同角色， 我的初步数据表明，双壬基修饰可以单独存在于K4 me 3中， 与具有推定结合复合物的H3 K4 me 3Q 5ser相比是独特的（例如，NuRD）。这项建议 有望建立重要的转录可塑性模式与组蛋白乙酰化， 个脑袋此外，该奖项提供的额外培训和科学环境将理想地 准备我启动一个独立的研究计划，在多个层次上进行评估（表观基因组， 细胞/行为），以了解组蛋白乙酰化如何有助于可塑性的正常和异常状态。