Neutrophil heterogeneity and plasticity in wound healing

项目总结/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功能， 组织和生物医学背景。我们将高维和单细胞分析与集成 生物信息学方法来定义上下文特异性中性粒细胞景观。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中起着至关重要的第一线作用， 病理反应。新出现的证据表明，中性粒细胞反应是系统性的， 具有独特的表型和功能。烧伤是最常见的创伤性损伤 国际吧在严重的情况下，患者可能死于感染、休克和/或器官衰竭，需要迅速的治疗。 临床反应。适当的伤口愈合需要协调的时间依赖性平衡的亲和抗- 炎症免疫途径，以防止不可逆的全身性损伤。尽管是第一个动员起来 损伤部位的中性粒细胞的异质性，其命运，以及哪些因素控制，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结构 在伤口愈合过程中随着时间的推移而变化，血液中性粒细胞可以作为预测 严重烧伤的并发症和结局。为此，我们将重点介绍两个重要的发现， 过去十年中性粒细胞生物学：（i）中性粒细胞异质性和对特定组织环境的适应 通过高维流式细胞术和单细胞转录组数据揭示，和（ii）中性粒细胞逆转录酶 迁移表型，在模型生物体中显示，证明中性粒细胞迁移回血管系统 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 中性粒细胞及其与其他免疫细胞的相互作用在人类烧伤使用多模式单细胞分析 方法，2）通过进行跨种属研究， 使用临床样本和预处理样本进行伤口愈合中性粒细胞异质性的单细胞转录组分析 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来定义， 小鼠模型。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， 可塑性的伤口愈合使用多学科的方法。我们的方法是创新的，因为我们将 采用：（i）最先进的单细胞技术，（ii）新的综合生物信息学方法开发， 分析，和（iii）使用模型生物体，用体内验证模型分析人体中跨组织的数据。 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景观 研究创伤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.