Mechanisms of endothelial regeneration and resolution of lung vascular injury

内皮再生机制和肺血管损伤的解决

• 批准号: 9893019
• 负责人: DOLLY MEHTA
• 金额: $ 51.97万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9893019
• 关键词: Address; Adult; Adult Respiratory Distress Syndrome; Binding; Blood Vessels; Cell Lineage; Cell Nucleus; Cells; Color; Data; Development; Endothelial Cells; Endothelium; Endotoxins; Family; Fibroblasts; Fluid Balance; Generations; Genes; Genetic; Genetic Transcription; Impairment; Inflammatory; Injury; Knockout Mice; Link; Lung; Lung diseases; Measurement; Mediating; Morbidity - disease rate; Mus; Natural regeneration; PTEN gene; Pathogenesis; Pathway interactions; Population; Protein Dephosphorylation; Pulmonary Edema; Recovery; Regulation; Reporter; Research; Resolution; Role; Sepsis; Signal Transduction; Source; Specific qualifier value; Tamoxifen; Testing; Time; Tissues; Trauma; Tumor Suppressor Proteins; Vascular Endothelium; Vascular Permeabilities; Vascularization; base; cell regeneration; effective therapy; endothelial regeneration; experimental study; lung injury; lung vascular injury; migration; mortality; mouse model; novel; novel therapeutic intervention; nucleocytoplasmic transport; preservation; prevent; programs; recruit; repaired; restoration; tissue injury; transcription factor; transcriptome sequencing

Project Summary. Loss of endothelial barrier integrity is central to pathogenesis of acute respiratory distress syndrome (ARDS), a severe lung disease, associated with sepsis and trauma. Thus, homeostatic replenishment of ECs after injury is required for the efficient formation of a stable endothelial barrier and to prevent long-lasting lung vascular injury but the identity and source of these “reparative endothelial cells (ECs)” remains unclear. Given the findings that ECs can convert into fibroblast-like cells (FLC) upon tissue injury and that forced expression of the ETS family of transcription factors (e.g., ETV2) reprogram FLC into ECs, a fundamental unanswered question is whether a shift from the “reparative” EC lineage into a non-reparative FLC lineage impairs endothelial regeneration and thereby recovery from lung vascular injury. PTEN (Phosphatase and TENsin homolog) is a well-known tumor suppressor and regulates key features of vascularization such as endothelial migration and proliferation. Our Preliminary Data, showed that EC-specific conditional deletion of PTEN markedly decreased lung EC regeneration while FLCs were increased and these mice developed lung edema spontaneously. Further experiments showed that PTEN localized to the nucleus of ECs and maintained the expression of endothelial transcription factor, Ets-Related Gene (ERG). Based on these intriguing Preliminary Data, and using a range of powerful approaches such as novel genetic mouse models (inducible dual-reporter EC-PTEN and EC-ERG null mice for lineage tracing), RNAseq and measurements of lung vascular permeability we will investigate the hypothesis that PTEN-induced ERG pathway is a key determinant of the EC lineage and resolution of lung-fluid balance. Our Specific Aims are: #1: to investigate the hypothesis that EC-expressed PTEN drives endothelial cell regeneration at the expense of fibroblast-like cells and thereby maintains the lung vascular barrier; #2: to determine (a) the role of the endothelial transcription factor, ERG, as an effector of PTEN and a key determinant of EC lineage, and (b) to identify ERG enriched endothelial cells through endothelial lineage tracing as intrinsic reparative ECs whose activation promotes lung vascular regeneration and restoration of lung fluid balance. With these comprehensive studies, we will identify for the first time the role of the PTEN-ERG cascade in maintaining the reparative EC lineage and in resolving inflammatory lung vascular injury. We expect our studies to lay a new conceptual framework for the development of novel therapeutic approaches exploiting PTEN as a key target for the treatment of ARDS.

项目摘要。内皮屏障完整性的丧失是急性呼吸窘迫发病机制的核心 综合症（ARDS），一种严重的肺部疾病，与败血症和创伤有关。因此，体内平衡补充 损伤后 ECs 的形成对于有效形成稳定的内皮屏障和防止长期持续的损伤是必需的。 肺血管损伤，但这些“修复性内皮细胞（EC）”的身份和来源仍不清楚。 鉴于 EC 可以在组织损伤时转化为成纤维细胞样细胞 (FLC) 的发现，这迫使 ETS 转录因子家族（例如 ETV2）的表达将 FLC 重新编程为 EC，这是一个基本的 未解答的问题是是否从“修复性”EC 谱系转变为非修复性 FLC 谱系 损害内皮再生，从而损害肺血管损伤的恢复。 PTEN（磷酸酶和 TENsin 同源物）是一种众所周知的肿瘤抑制因子，可调节血管化的关键特征，例如 内皮迁移和增殖。我们的初步数据表明，EC 特定的条件删除 PTEN 显着降低了肺 EC 再生，而 FLC 增加，这些小鼠出现了肺 自发性水肿。进一步的实验表明，PTEN定位于ECs的细胞核并维持 内皮转录因子 Ets 相关基因 (ERG) 的表达。基于这些有趣的 初步数据，并使用一系列强大的方法，例如新型遗传小鼠模型（诱导型 双报告基因 EC-PTEN 和 EC-ERG 缺失小鼠用于谱系追踪）、RNAseq 和肺血管测量 我们将研究以下假设：PTEN 诱导的 ERG 通路是 EC 的关键决定因素 肺液平衡的谱系和分辨率。我们的具体目标是： #1：调查以下假设： EC表达的PTEN以成纤维细胞样细胞为代价驱动内皮细胞再生 从而维持肺血管屏障； #2：确定 (a) 内皮转录的作用 因子 ERG 作为 PTEN 的效应子和 EC 谱系的关键决定因素，以及 (b) 鉴定 ERG 富集 内皮细胞通过内皮谱系追踪作为内在修复性 EC，其激活 促进肺血管再生和恢复肺液平衡。有了这些全面的 研究中，我们将首次确定 PTEN-ERG 级联在维持修复性 EC 中的作用 谱系和解决炎症性肺血管损伤。我们期望我们的研究能够提出一个新的概念 利用 PTEN 作为治疗关键靶点的新型治疗方法的开发框架 急性呼吸窘迫综合征。

项目成果

Protease-activated receptor 2 promotes clearance of Pseudomonas aeruginosa infection by inducing cAMP-Rac1 signaling in alveolar macrophages.

• DOI: 10.3389/fphar.2022.874197
• 发表时间: 2022
• 期刊: FRONTIERS IN PHARMACOLOGY
• 影响因子: 5.6
• 作者: Rayees, Sheikh;Joshi, Jagdish Chandra;Joshi, Bhagwati;Vellingiri, Vigneshwaran;Banerjee, Somenath;Mehta, Dolly
• 通讯作者: Mehta, Dolly