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

项目总结 人类的健康深受遗传和环境的影响。社会环境起着中介作用 生理、基因调节和免疫信号的变化，导致疾病的显著差异 易感性和死亡率。长期的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 对感染性病原体免疫反应的影响。然而，这些现象背后的分子机制 人们对其影响知之甚少。为了解决这一差距，强大的实验模型揭示了社会 环境对免疫系统的影响是必要的。 猕猴是与人类关系密切的灵长类动物。在社会群体中，他们保持稳定 社会支配地位的等级制度，这些等级制度可以通过社会团体实验性地操纵 重新编排。因此，它们为研究疾病的因果社会等级效应提供了独特的资源。虽然 已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描述。单细胞RNA测序 能够发现社会环境对多种个体细胞的影响，这有助于 靶向免疫失调的与临床相关的细胞类型。因此，这项工作 建议研究个体外周血细胞在基线和治疗后对基因调控的等级效应 免疫刺激，揭示特定细胞类型的社会应激对免疫的影响。 推动这一提议的基础研究问题是：社会环境如何影响基因 在不同的外周血液免疫细胞中表达？为了回答这个问题，这项提议的方法 是在圈养雌性猕猴身上使用一个成熟的社会逆境模型，并应用下一步- 新一代单细胞RNA-seq技术，用于深入鉴定从50个个体中分离出来的细胞 社会分化。根据这些数据，将检查社会等级的影响：(1)免疫成分和内毒素- 激发极化，(2)跨细胞类型的基因表达稳定性，以及(3)基因调控网络和 共表达模块。通过这项工作，与临床相关的基因和途径将被识别出来 根据社会环境影响重新布线，以设计靶向疗法来改善免疫反应 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.