Quantifying gene expression and network regulation in single cells to reveal the consequences of stress on the immune response

项目摘要 人类健康受到遗传和环境的深刻影响。社会环境起中介作用 生理学、基因调控和免疫信号的变化，导致疾病的显著差异 易感性和死亡率。慢性社会压力和社会不平等是疾病的主要驱动因素，部分原因是 对感染性病原体的免疫反应的影响。然而，这些潜在的分子机制 效果知之甚少。为了解决这一差距，强大的实验模型，揭示社会 需要环境对免疫系统的影响。 恒河猴是与人类关系密切的灵长类动物。在社会群体中， 社会统治的等级，这些等级可以通过社会群体实验操纵 重排因此，他们提供了一个独特的资源，研究因果关系的社会地位对疾病的影响。虽然 已经发现外周血中显著和实质性的基因表达变化与 社会等级，这些签名背后的细胞类型没有得到很好的表征。单细胞RNA测序 能够发现社会环境对多种细胞的影响，这有助于 靶向导致免疫失调的临床相关细胞类型。因此，这项工作 建议在基线和基线后研究个体外周血细胞中基因调控的等级效应， 免疫刺激，以揭示细胞类型特异性社会应激对免疫力的影响。 推动这一提议的基本研究问题是：社会环境如何影响基因 在不同的外周血免疫细胞中表达？为了回答这个问题，本提案的方法 是在圈养的雌性恒河猴中使用一个成熟的社会逆境模型，并应用下一个- 一代单细胞RNA-seq技术，以深入表征从50个个体中分离的细胞， 社会梯度根据这些数据，将检查社会等级的影响：（1）免疫组成和LPS- 诱导极化，（2）跨细胞类型的基因表达稳健性，和（3）基因调控网络， 共表达模块。通过这项工作，临床相关的基因和途径将被确定， 通过社会环境的影响重新连接，以设计有针对性的治疗方法来改善免疫反应 与癌症和传染病有关。

PROJECT SUMMARY Human health is profoundly affected by genetics and the environment. The social environment mediates changes in physiology, gene regulation, and immune signaling, resulting in significant differences in disease susceptibility and mortality. Chronic social stress and social inequity are major drivers of disease, in part due to effects on the immune response to infectious pathogens. However, the molecular mechanisms underlying these effects are poorly understood. In order to address this gap, robust experimental models for uncovering social environmental effects on the immune system are needed. Rhesus macaques are primates closely related to humans. In social groups, they maintain stable hierarchies of social dominance, and these hierarchies can be experimentally manipulated through social group rearrangement. Thus, they provide a unique resource to study causal social rank effects on disease. Although significant and substantial gene expression changes in peripheral blood have been found to be associated with social rank, the cell types underlying these signatures are not well-characterized. Single-cell RNA-sequencing enables the discovery of effects of the social environment on individual cells of many kinds, which can help with the targeting of clinically-relevant cell types that are responsible for immune dysregulation. Therefore, this work proposes to study rank effects on gene regulation in individual peripheral blood cells, at baseline and after immune stimulation, to uncover cell-type-specific social stress effects on immunity. The basic research question driving this proposal is: How does the social environment affect gene expression across diverse peripheral blood immune cells? To answer this question, the approach of this proposal is to use a well-established model of social adversity in captive female rhesus macaques, and to apply next- generation single-cell RNA-seq technology to deeply characterize cells isolated from 50 individuals across a social gradient. From this data, effects of social rank will be examined for: (1) immune composition and LPS- induced polarization, (2) gene expression robustness across cell types, and (3) gene regulatory networks and co-expression modules. Through this work, clinically-relevant genes and pathways will be identified that are rewired by social environment effects, in order to design targeted therapeutics to improve immune responses relevant to cancer and infectious disease.