Mechanisms of regulatory T-cell mediated endothelial repair following viral pneumonia in aged hosts

老年宿主病毒性肺炎后调节性 T 细胞介导的内皮修复机制

• 批准号: 10687164
• 负责人: Luisa Morales-Nebreda
• 金额: $ 16.12万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-20 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10687164
• 关键词: 2019-nCoV; Acute Lung Injury; Acute Respiratory Distress Syndrome; Adoptive Transfer; Affect; Age; Aging; Alveolar; Angiogenic Factor; Antiviral Therapy; Autopsy; Biological Process; Blood Vessels; Blood capillaries; Bronchoalveolar Lavage Fluid; Case/Control Studies; Cause of Death; Cell Therapy; Cells; Clinical; Data; Dedications; Development; Diffuse; Down-Regulation; Elderly; Endothelial Cells; Endothelium; Experimental Designs; Failure; Flow Cytometry; Fluorescence-Activated Cell Sorting; Functional disorder; Gene Expression Profile; Gene Expression Profiling; Genetic Transcription; Goals; Human; Immune; Impairment; Inflammation; Influenza; Influenza A virus; Injury; Knowledge; Learning; Link; Longevity; Lung; Lymphocytic Infiltrate; Measurement; Mediating; Mentors; Methods; Molecular; Mus; Older Population; Organ; Outcome; Pathology; Patients; Phase; Phenotype; Physicians; Pneumonia; Population; Predisposition; Printing; Production; Proteins; Publishing; Pulmonary Inflammation; Recovery; Regulatory T-Lymphocyte; Research; Resolution; Risk Factors; Scientist; Severities; Specimen; Techniques; Testing; Thrombosis; Training; Transgenic Organisms; United States; VEGFA gene; Vascular Endothelial Growth Factors; Viral Pneumonia; Virus; Virus Diseases; Youth; adaptive immune response; age related; aged; alveolar epithelium; angiogenesis; career; cell age; design; endothelial repair; experience; experimental study; gain of function; improved; influenza infection; influenzavirus; insight; loss of function; lung injury; lung repair; mortality; mouse model; novel; older patient; overexpression; programs; recruit; repair function; repaired; response to injury; skills; small molecule; tissue repair; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT The broad objectives of this K08 proposal are two-fold: 1) to facilitate development of the necessary skills that will allow the candidate to achieve her long-term goal of becoming a successful physician-scientist focusing on determinants of recovery from viral pneumonia in aged hosts, and 2) to investigate the mechanisms that direct the Tregs to orchestrate resolution of severe lung injury throughout the lifespan. The candidate and her mentors have designed a detailed training plan tailored to the candidate's specific needs and goals. The plan includes a rigorous research component that will afford the PI new knowledge and research skills to better examine the links between a critical immune cell population and the endothelium during recovery from viral pneumonia. The proposal concerns viral pneumonia, specifically influenza-induced lung injury, its clinical counterpart the acute respiratory distress syndrome (ARDS) and how they both disproportionately affect the elderly population. Despite decades of dedicated research, there are only a few anti-viral therapies with limited efficacy to manage severe viral pneumonia. Tregs have been shown to decrease inflammation and promote tissue repair in diverse murine models of lung injury. Tregs also increase in the lungs of patients with ARDS, suggesting that they may play a role in the human adaptive immune response to lung injury. However, the specific mechanisms that cause Tregs to execute their pro-repair program following lung injury remain unknown. Our preliminary data shows that the youthful reparative Treg cell program following influenza-induced lung injury is dominated by biologic processes linked to the development and repair of blood capillaries. This reparative program is lost in aged hosts in a cell- autonomous manner. Thus, we hypothesize that aging results in Treg cell-specific downregulation of important pro-angiogenic factors expression such as VEGFA, leading to impaired alveolar endothelial repair and recovery from viral pneumonia. The long-term goal of the proposal is to identify novel small molecule- and cell-based therapeutics to control inflammation and promote tissue repair in our increasingly older population. To test this hypothesis, we propose the following Specific Aims: 1) determine whether age-related alterations in the pro- endothelial repair function of Tregs results from cell-autonomous or microenvironmentally-driven changes, 2) determine whether Treg cell-generated VEFGA is necessary and sufficient to restore the pro-endothelial repair function present in youth that is lost with aging, and 3) determine whether age-related VEGFA expression in alveolar Tregs in bronchoalveolar lavage fluid is associated with 30-day mortality in patients with severe viral pneumonia. We will use standard techniques to assess severity of lung injury, endothelial repair, heterochronic (age mis-matched) adoptive Treg cell transfer, multiple transgenic murine strains for loss-of-function and gain- of-function experiments, flow cytometry, fluorescence-activated cell sorting and transcriptional profiling with RNA-sequencing as the primary methods to support the experimental design of this proposal.

项目总结/摘要 K 08提案的主要目标有两个方面：1）促进必要技能的发展， 将使候选人实现她的长期目标，成为一个成功的医生，科学家，专注于 老年宿主病毒性肺炎恢复的决定因素，2）研究指导 在整个生命周期中协调解决严重肺损伤的Tendon。候选人和她的导师 根据候选人的具体需求和目标设计了详细的培训计划。该计划包括 严格的研究组成部分，将提供PI新的知识和研究技能，以更好地检查 病毒性肺炎恢复期间关键免疫细胞群与内皮之间的联系。的 建议涉及病毒性肺炎，特别是流感引起的肺损伤，其临床对应的急性 呼吸窘迫综合征（ARDS）以及它们如何不成比例地影响老年人。尽管 经过几十年的专门研究，只有少数疗效有限的抗病毒疗法可以治疗严重的 病毒性肺炎在不同的小鼠中，已显示TGFAP可减少炎症并促进组织修复。 肺损伤模型。在ARDS患者的肺中，甲状腺激素也会增加，这表明它们可能在ARDS中起着重要作用。 在人类对肺损伤的适应性免疫反应中的作用。然而，导致Tyndrome的具体机制 在肺损伤后执行其亲修复程序仍然未知。初步数据显示， 流感诱导的肺损伤后年轻的修复性Treg细胞程序受生物过程支配 与毛细血管的发育和修复有关。在细胞中，这个修复程序在年老的宿主中丢失了- 自主的方式。因此，我们假设衰老导致Treg细胞特异性下调重要的免疫调节因子。 促血管生成因子如VEGFA的表达，导致肺泡内皮修复和恢复受损 病毒性肺炎该提案的长期目标是确定新的小分子和细胞为基础的 治疗控制炎症和促进组织修复在我们日益老龄化的人口。为了验证这一 假设，我们提出了以下具体目标：1）确定是否与年龄相关的变化，在前， 内皮细胞修复功能由细胞自主或微环境驱动的变化引起，2） 确定Treg细胞产生的VEFGA是否是恢复促内皮修复所必需和足够的 功能存在于年轻人中，随着年龄的增长而丧失，和3）确定是否与年龄相关的VEGFA表达在年轻人中， 支气管肺泡灌洗液中的肺泡炎与重症病毒性肺炎患者的30天死亡率相关 肺炎我们将使用标准技术评估肺损伤的严重程度、内皮修复、异时性 (age不匹配）过继性Treg细胞转移，多种转基因小鼠品系用于功能丧失和获得- 功能实验、流式细胞术、荧光激活细胞分选和转录谱分析， 以RNA测序为主要方法支持本方案的实验设计。