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

CXCL12 信号轴在肺动脉异质性、发育和疾病中的作用

• 批准号: 10905162
• 负责人: David Brian Frank
• 金额: $ 65.73万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-12 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10905162
• 关键词: Adult; Angiogenesis Pathway; Arteries; Autoimmune Diseases; Automobile Driving; Biological Assay; Blood Vessels; Blood capillaries; CXCR4 gene; Cell surface; Cells; Cellular Assay; Child; Childhood; Chronic Disease; Congenital diaphragmatic hernia; Data; Development; Differentiation and Growth; Disease; Distal; Endothelial Cells; Endothelium; Etiology; Fluorescence-Activated Cell Sorting; Generations; Genes; Genetic; Growth; Heterogeneity; Homeostasis; Hypoxemia; In Vitro; Injury; Label; Lead; Left; Longevity; Lung; Lung diseases; Maintenance; Malignant Neoplasms; Mediator; Methods; Modeling; Molecular; Molecular Target; Mus; Natural regeneration; Ontology; Oxygen; Paper; Pathogenesis; Pathology; Pathway interactions; Pattern; Population; Prevention; Proliferating; Pulmonary Circulation; Pulmonary Hypertension; Pulmonary artery structure; Regulation; Reporter; Role; Series; Signal Transduction; Stromal Cell-Derived Factor 1; Structure of parenchyma of lung; System; Technology; Testing; Tissue Differentiation; Tissues; Trees; Vascular Endothelial Cell; Vascular resistance; Vascularization; Vasodilator Agents; angiogenesis; biobank; cell motility; cellular targeting; disorder prevention; fetal; genetic approach; genetic manipulation; improved; in utero; lung development; lung injury; migration; morphogens; mouse model; nitrofen; novel; novel therapeutics; pharmacologic; programs; pulmonary vascular disorder; receptor; recruit; red fluorescent protein; repaired; response to injury; right ventricular failure; segregation; single cell technology; single-cell RNA sequencing; spatiotemporal; stem cells; targeted treatment; therapeutic target; therapy design; transcriptomics; vascular bed

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.

项目摘要 发育范式的重新接触经常指示疾病发病机理。虽然至关重要 组织形成的生长和分化，这些途径的异常表达导致显着 癌症，自身免疫性和慢性疾病等病理。同样，在正常胎儿计划中的破坏 肺在发育和失去体内平衡期间会导致明显的肺血管发育不全 成年人，表明在预防疾病中起着共同的作用。因此，了解细胞和分子 策划肺血管发育的机制将有助于确定肺部疾病的治疗靶标 为了研究驱动发育的细胞和分子途径，我们专注于动脉 内皮，这是一种对调节血管耐药性和预防肺部的血管室 疾病这种肺动脉高压（pH）。我们使用CXCL12DSRED标记了动脉内皮细胞（EC） 荧光记者小鼠，其中在表达标志动脉的细胞中产生红色荧光蛋白 基因，CXCL12。我们分离了CXCL12+细胞，并进行了单细胞RNA测序以评估细胞 肺部内皮中的异质性。组合细胞注释，基因本体分析和空间 转录组学在发育中的动脉EC上揭示了空间和功能上不同的新型亚群 肺。此外，我们发现了从动脉到毛细血管的CXCL12形态梯度，这表明是 肺动脉生长的触觉机制。此外，导致形态梯度的破坏 在我们通过应用独特的方法来量化的肺血管性发育不全和异常分支中 肺。 在此提案中，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的转录组学促进了我们的动脉生长的假设 组装由CXCL12形态梯度指导毛细管的细胞迁移控制 祖细胞到远端动脉。此外，这种表达模式被观察到成年，我们 假设肺动脉树包含空间和功能上不同的亚群 在整个生命周期中保守。我们将使用遗传小鼠模型和单一测试这些假设 细胞技术以发现动脉异质性和CXCL12的肺血管的机制 发展。 目的1。定义CXCL12信号传导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性EC的异质性和信号传导。 目标3。识别和表征进化保守的近端和远端动脉EC。