Neutrophil heterogeneity and plasticity in wound healing

项目摘要/摘要 我的研究计划的长期目标是了解中性粒细胞的异质性和在不同的 组织和生物医学背景。我们将高维和单细胞分析与集成 生物信息学方法确定特定背景的中性粒细胞景观。中性粒细胞，最丰富的 人类免疫细胞类型，在调节对感染的快速反应和 病理性反应。新出现的证据表明，中性粒细胞的反应是系统性的和背景的- 特有的，具有不同的表型和功能。烧伤是最常见的创伤之一。 全世界。在严重情况下，患者可能死于感染、休克和/或器官衰竭，需要迅速 临床反应。适当的伤口愈合需要协调一致的时间依赖平衡的支持和反对 炎性免疫途径，防止不可逆的全身损害。尽管是第一个动员到 对于中性粒细胞的异质性、它们的命运以及哪些因素控制，人们仍然知之甚少。 中性粒细胞在伤口愈合中的可塑性。我们假设中性粒细胞的细胞和分子景观 在伤口愈合过程中随着时间的变化，血液中的中性粒细胞可能是一个预测的标志 严重烧伤的并发症和转归。为此，我们将重点关注以下两个重要发现 过去十年的中性粒细胞生物学：(I)中性粒细胞的异质性和对特定组织环境的适应 高维流式细胞术和单细胞转录组数据显示：(Ii)中性粒细胞逆转 迁移表型，在模型生物中显示中性粒细胞迁移回血管系统 作为对炎症的反应。在这个Mira应用程序中，我计划1)识别转录和空间景观 应用多模式单细胞图谱研究烧伤患者中性粒细胞及其与其他免疫细胞的相互作用 方法，2)通过跨物种研究确定烧伤患者中性粒细胞的反向迁移表型 单细胞转录组分析伤口愈合过程中中性粒细胞的异质性 临床斑马鱼烧伤模型，以及3)开发预测中性粒细胞行为的分析管道和方法 通过使用人类单细胞基因组数据和斑马鱼的公开数据进行成像定义 老鼠模型。在接下来的五年里，我将建立一个研究项目，研究中性粒细胞的异质性和 用多学科方法研究伤口愈合中的可塑性。我们的方法是创新的，因为我们将 使用：(一)最先进的单细胞技术，(二)新的综合生物信息学方法的开发和 分析，以及(Iii)通过使用模型生物体的活体验证模型来分析人类的跨组织数据。 这些研究将产生全面的数据，并提供中性粒细胞的细胞和分子图景 研究伤口修复和中性粒细胞生物学的社区的异质性。

PROJECT SUMMARY/ABSTRACT The long-term goal of my research program is to understand neutrophil heterogeneity and functions in diverse tissues and biomedical contexts. We have combined high-dimensional and single-cell profiling with integrated bioinformatics approaches to defining context-specific neutrophil landscape. Neutrophils, the most abundant human immune cell type, play crucial, first-line roles in regulating swift responses against infections and pathological responses. Emerging evidence has shown that neutrophil response is systematic and context- specific with distinct phenotypes and functions. Burn injuries are among the most common traumatic injuries worldwide. In severe cases, patients can die of infection, shock, and/or organ failure, requiring expeditious clinical responses. Proper wound healing requires a coordinated time-dependent balance of pro- and anti- inflammatory immune pathways to prevent irreversible systemic damage. Despite being the first to mobilize to sites of injury, there remains little known about neutrophil heterogeneity, their fates, and which factors control the plasticity of neutrophils in wound healing. We hypothesize that neutrophils' cellular and molecular landscape change over time during the wound healing process, and that blood neutrophils could be a marker to predict complications and outcomes in severe burn injury. To this end, we will focus on two important discoveries in neutrophil biology over the past decade: (i) neutrophil heterogeneity and adaption to specific tissue environments revealed by high-dimensional flow cytometry and single-cell transcriptome data and (ii) neutrophil reverse migration phenotypes, shown in model organisms demonstrating that neutrophils migrate back to the vasculature in response to inflammation. In this MIRA application, I plan to 1) identify transcriptional and spatial landscapes of neutrophils and their interactions with other immune cells in human burns using multimodal single-cell profiling approaches, 2) define reverse migration neutrophil phenotypes in human burns by performing cross-species single-cell transcriptome analyses of neutrophil heterogeneity in wound healing using clinical samples and pre- clinical zebrafish burn models, and 3) develop analysis pipelines and methods to predict neutrophil behaviors defined by imaging using single-cell genomic data in humans and publicly available data from zebrafish and mouse models. In the next five years, I will establish a research program to study neutrophil heterogeneity and plasticity in wound healing using multidisciplinary approaches. Our approaches are innovative because we will employ: (i) state-of-the-art single-cell technologies, (ii) novel integrated bioinformatics method development and analysis, and (iii) profiling data cross-tissues in humans with in vivo validation models using model organisms. These studies will generate comprehensive data and provide cellular and molecular landscapes of neutrophil heterogeneity to the community studying wound repair and neutrophil biology.