Epigenetic and cellular markers of stress sensitization by early life stress in mice

修改后的项目摘要/摘要部分 这个项目将在普林斯顿大学的普林斯顿神经科学研究所继续进行。早期生活压力(ELS)是导致抑郁症和其他精神疾病的最强烈的终生风险因素之一，特别是在成年后面临额外的压力事件之后。然而，对于发育中的大脑如何编码ELS的经验以增加对额外压力的敏感性，人们知之甚少。为了解决这个问题，我们建立了一个翻译上相关的早期生活和成年应激小鼠模型，在该模型中，在特定的出生后敏感窗口期间的应激增加了成年后应激导致抑郁样行为的可能性。这项建议将检查转录、表观遗传调节和细胞微电路重新激活的变化是否由ELS经验诱导，以及这些变化是否直接导致应激敏感型抑郁样行为。在目标1和目标3中，我们将使用各种尖端技术，包括RNA测序、染色质免疫沉淀和ATAC测序，研究表观遗传格局的转录和“启动”的变化。在目标2中，我们将使用活动依赖转基因小鼠(ArcCreERT2)来研究ELS激活的中皮质边缘奖赏相关脑区的细胞是否在成年应激下重新激活。在目标3中，我们将使用新的转基因杂交(ArcCreERT2 x R26-CAGLSL-SUN1-sfGFP-myc)将这些方法结合起来，首次检查从ELS激活的细胞分离的核内表观遗传启动的变化。然后，我们将使用药物遗传抑制来评估ELS最初激活细胞以用于随后的应激敏化和抑郁样行为的必要性。此外，我们将使用CRISPR构建物来敲除Setd7，这是一种通过H3K4me1沉积建立表观遗传启动的酶。这将使我们能够测试这个标记与ELS的应激敏化效应的功能相关性。总而言之，这项在《独立之路奖》中提出的研究将揭示ELS诱导的长期应激敏化和增强抑郁和精神疾病易感性的独立和潜在的相互作用的分子和细胞机制。

Modified Project Summary/Abstract Section This project will be continued at Princeton University in the Princeton Neuroscience Institute. Early life stress (ELS) is one of the strongest lifetime risk factors for developing depression and other psychiatric disorders, particularly after facing additional stressful events in adulthood. However, relatively little is known about how the developing brain encodes experience of ELS to increase sensitivity to additional stress. To address this, we have established a translationally relevant “two-hit” mouse model of early life and adult stress, wherein stress during a specific postnatal sensitive window increases the likelihood that stress in adulthood will lead to depressive-like behaviors. This proposal will examine whether changes in transcription, epigenetic regulation, and cellular microcircuit reactivation are induced by ELS experience, and whether these changes directly contribute to stress-sensitized depression-like behaviors. In Aims 1 and 3, we will investigate changes in transcription and “priming” of the epigenetic landscape using a variety of cutting-edge techniques including RNA sequencing, chromatin immunoprecipitation, and ATAC-sequencing. In Aim 2, we will ask whether cells in mesocorticolimbic reward-related brain regions activated by ELS are reactivated by adult stress, using activity dependent transgenic mice (ArcCreERT2). In Aim 3 we will then combine these approaches using a novel transgenic cross (ArcCreERT2 x R26-CAGLSL- Sun1-sfGFP-myc) to examine, for the first time, changes in epigenetic priming specifically within nuclei isolated from ELS-activated cells. We will then use pharmacogenetic inhibition to assess the necessity of cells initially activated by ELS for subsequent stress sensitization and depression-like behavior. In addition, we will use CRISPR constructs to knock out Setd7, an enzyme that establishes epigenetic priming by H3K4me1 deposition. This will allow us to test the functional relevance of this mark for the stress sensitization effects of ELS. In sum, the research proposed in this Pathway to Independence Award will reveal both separate and potentially interactive molecular and cellular mechanisms of long-lasting ELS-induced stress sensitization and enhanced vulnerability to depression and psychiatric disease.