CXCL12 signaling axis in pulmonary arterial heterogeneity, development, and disease

项目总结 发育范式的重新结合经常指导疾病的发病机制。虽然对 生长和分化为组织形成，这些途径的异常表达导致显著 病理学，如癌症、自身免疫和慢性病。同样，在美国，正常胎儿计划的中断 肺在发育过程中可导致明显的肺血管发育不良和内稳态丧失。 成人，表明在预防疾病中起着共同的作用。因此，理解细胞和分子 协调肺血管发育的机制将有助于确定肺部疾病的治疗靶点 为了研究推动发育的细胞和分子途径，我们将重点放在动脉上。 血管内皮细胞在调节血管阻力和预防肺损伤中的重要作用 疾病，如肺动脉高压(PH)。我们用Cxcl12DsRed标记动脉内皮细胞 一种荧光报告鼠，在表达标志动脉的细胞中产生红色荧光蛋白 吉恩，CXCL12。我们分离CXCL12细胞并进行单细胞RNA测序以评估细胞 肺内皮细胞的异质性。结合了细胞注释、基因本体分析和空间 转录组学揭示了发育中动脉内皮细胞在空间和功能上截然不同的新亚群 阿龙。此外，我们还发现了从动脉到毛细血管的CXCL12形态梯度，表明 对肺动脉生长的趋触性机制。此外，形态原梯度的破坏导致 在肺血管发育不良和异常分支中，我们通过应用独特的方法对 阿龙。 在这项提案中，我们正在评估CXCL12在发育和疾病中的扩大作用。空间 CXCL12及其受体CXCR4和Ackr3的转录转录增强了我们的假设，即动脉生长 和组装由CXCL12形态原梯度控制，指导毛细血管的细胞迁移 远端动脉的祖细胞。此外，这种表达模式会一直观察到成年，而我们 假设肺动脉树包含空间和功能上不同的亚群 在整个生命周期中得到保护。我们将使用遗传小鼠模型和单个 细胞技术揭示动脉异质性和CXCL12协调的肺血管的机制 发展。 目的1.明确CXCL12信号在肺血管发育中的时空作用。 目的2.评估发育性肺部疾病中依赖CXCL12的EC的异质性和信号转导。 目的3.鉴定和表征进化上保守的近端和远端动脉内皮细胞。

PROJECT SUMMARY Reengagement of developmental paradigms frequently instructs disease pathogenesis. While critical for growth and differentiation for tissue formation, aberrant expression of these pathways leads to significant pathology such as cancer, autoimmunity, and chronic disease. Similarly, disruption of normal fetal programs in the lung can result in significant pulmonary vascular hypoplasia during development and loss of homeostasis in the adult, suggesting a common role in prevention of disease. Thus, understanding the cellular and molecular mechanisms orchestrating pulmonary vascular development will help identify therapeutic targets for lung disease To investigate the cellular and molecular pathways driving development, we focused on the arterial endothelium, a vascular compartment important in the regulation of vascular resistance and prevention of lung diseases such pulmonary hypertension (PH). We labeled arterial endothelial cells (ECs) using the Cxcl12DsRed fluorescent reporter mouse in which a red fluorescent protein is produced in cells expressing the hallmark arterial gene, Cxcl12. We isolated Cxcl12+ cells and performed single cell RNA sequencing to assess cellular heterogeneity within the pulmonary endothelium. Combined cell annotation, gene ontology analysis, and spatial transcriptomics revealed spatially and functionally distinct novel subpopulations of arterial ECs in the developing lung. In addition, we discovered a CXCL12 morphogen gradient from arteries to capillaries, suggesting a haptotaxis mechanism to pulmonary artery growth. Furthermore, disruption of the morphogen gradient resulted in pulmonary vascular hypoplasia and aberrant branching that we quantified by applying unique methods for the lung. In this proposal, we are assessing an expanded role for CXCL12 in development and disease. Spatial transcriptomics for Cxcl12 and its receptors, Cxcr4 and Ackr3, promote our hypothesis that arterial growth and assembly is governed by a CXCL12 morphogen gradient directing cell migration of capillary progenitor cells to the distal artery. In addition, this pattern of expression is observed into adulthood, and we hypothesize that the pulmonary arterial tree contains spatially and functionally distinct subpopulations conserved throughout the lifespan. We will test these hypotheses using genetic mouse models and single cell technology to uncover mechanisms of arterial heterogeneity and CXCL12-orchestrated pulmonary vascular development. Aim 1. To define the spatiotemporal role of CXCL12 signaling in pulmonary vascular development. Aim 2. To assess CXCL12-dependent EC heterogeneity and signaling in developmental lung disease. Aim 3. To identify and characterize evolutionarily conserved proximal and distal arterial ECs.