Macrophage Reprogramming During Granuloma Formation in the Zebrafish

摘要 肉芽肿形成为对各种炎性和感染性刺激的保守宿主反应。AS 肉芽肿聚集，巨噬细胞交错排列，并经历显著的形态转变，呈现出 上皮样外观。这种转变的基础和对疾病的后果并不是很好 明白了。我们确定了巨噬细胞的保守重编程，它是组装和 分枝杆菌肉芽肿的稳定性。使用斑马鱼模型，我们发现广泛的上皮模块和 结构是在结核肉芽肿形成过程中形成的，对肉芽肿的完整性至关重要。在这 我们将1)评估特定的2型免疫信号如何与抗1型免疫信号相互作用以 协调上皮样转化和肉芽肿聚集；2)检测EMP2/局部黏附的作用 激酶(FAK)通路在肉芽肿稳定、解聚和感染扩散中的作用；3)基于 从单个动物分离的肉芽肿和巨噬细胞的scrna-seq分析，评估其作用 JAG1-Notch在肉芽肿形成、维持和感染轨迹中的作用。我们将从以下方面扩展调查结果 这些研究是对人类疾病的分析。总体而言，这一提议将检验这样一个假设，即在一个引人注目的 与发育和癌症中间充质到上皮的转变平行，巨噬细胞利用经典的 发生上皮样转变的发育信号通路。这种重新编程是 结核病的中心结构，并定义与宿主免疫系统的相互作用。我们将测试如何 这些途径的干扰会导致疾病进展和结果的改变。一种新的视角 这一关键结构可能对我们理解疾病的进展和 为新的治疗方法提供了机会。

Abstract Granulomas form as a conserved host response to a variety of inflammatory and infectious stimuli. As granulomas assemble, macrophages interdigitate and undergo a striking morphological transition, taking on an epithelioid appearance. The basis for this transformation and the consequences to disease are not well understood. We identified a conserved reprogramming of macrophages that underlies the assembly and stability of mycobacterial granulomas. Using a zebrafish model, we find that broad epithelial modules and structures are induced during tuberculous granuloma formation and are critical for granuloma integrity. In this project we will 1) assess how specific Type 2 immune signals interact with countervailing Type 1 signals to coordinate epithelioid transformation and granuloma assembly; 2) test the role of the EMP2/Focal Adhesion Kinase (FAK) pathway in granuloma stability, disaggregation and the dissemination of infection; 3) based on scRNA-seq analysis of granulomas as well as macrophages isolated from individual animals, assess the role of JAG1-Notch in granuloma formation, maintenance, and infection trajectory. We will extend findings from these studies into analysis of human disease. Overall, this proposal will test the hypothesis that, in a striking parallel to mesenchymal-to-epithelial transitions in development and cancer, macrophages draw on classical developmental signaling pathways to undergo an epithelial-like transition. This reprogramming underlies the central structure of tuberculosis and defines interactions with the host immune system. We will test how perturbations of these pathways lead to alterations in disease progression and outcome. A new perspective on this critical structure may have important implications for our understanding of disease progression and provides opportunities for new therapeutic approaches.