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

基本信息

批准号：
10283771
负责人：
Luisa Morales-Nebreda
金额：
$ 16.22万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
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/10283771
关键词：
2019-nCoV Acute Lung Injury Adoptive Transfer Adult Respiratory Distress Syndrome 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 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 Infiltration Inflammation Influenza Influenza A virus Injury Knowledge Learning Link Longevity Lung Lung Inflammation Lymphocyte Measurement Mediating Mentors Methods Molecular Target Mus Older Population Outcome Pathology Patients Phase Phenotype Physicians Play Pneumonia Population Predisposition Proteins Publishing Recovery Regulatory T-Lymphocyte Research Resolution Risk Factors Role Scientist Severities Specimen Structure Techniques Testing Thrombosis Training Transgenic Organisms United States VEGFA gene Viral Viral Pneumonia Virus Virus Diseases Youth adaptive immune response age related aged alveolar epithelium angiogenesis career cell age design 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

项目摘要

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.

项目概要/摘要 K08 提案的广泛目标有两个：1) 促进必要技能的发展，将使候选人能够实现成为一名成功的医师科学家的长期目标，重点关注老年宿主从病毒性肺炎中恢复的决定因素，2) 研究指导机制 Tregs 在整个生命周期中协调解决严重肺损伤。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。该计划包括一个严格的研究部分将为 PI 提供新的知识和研究技能，以便更好地检查病毒性肺炎恢复过程中关键免疫细胞群与内皮细胞之间的联系。这该提案涉及病毒性肺炎，特别是流感引起的肺损伤，其临床对应的急性肺损伤呼吸窘迫综合征 (ARDS) 以及它们如何对老年人群产生不成比例的影响。尽管经过数十年的专门研究，只有少数抗病毒疗法对治疗严重的病毒疗效有限病毒性肺炎。 Tregs 已被证明可以减少多种小鼠的炎症并促进组织修复肺损伤模型。 ARDS 患者肺部中的 Tregs 也有所增加，这表明它们可能发挥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Tregs的具体机制在肺损伤后执行他们的修复计划仍然未知。我们的初步数据表明流感引起的肺损伤后年轻的修复性 Treg 细胞程序由生物过程主导与毛细血管的发育和修复有关。这种修复程序在细胞中的衰老宿主中丢失了—— 自主方式。因此，我们假设衰老导致重要的 Treg 细胞特异性下调促血管生成因子表达，例如 VEGFA，导致肺泡内皮修复和恢复受损来自病毒性肺炎。该提案的长期目标是确定基于小分子和细胞的新型药物在日益老龄化的人群中控制炎症和促进组织修复的疗法。为了测试这个假设，我们提出以下具体目标：1）确定亲和力是否发生与年龄相关的变化。 Tregs 的内皮修复功能源于细胞自主或微环境驱动的变化，2) 确定 Treg 细胞生成的 VEFGA 是否对于恢复促内皮修复是必要且充分的年轻时存在的随着衰老而丧失的功能，3) 确定年龄相关的 VEGFA 表达是否在支气管肺泡灌洗液中的肺泡 Tregs 与重症病毒感染患者 30 天死亡率相关肺炎。我们将使用标准技术来评估肺损伤、内皮修复、异时性肺损伤的严重程度（年龄不匹配）过继性 Treg 细胞转移，多种转基因小鼠品系的功能丧失和增益功能实验、流式细胞术、荧光激活细胞分选和转录分析 RNA测序作为支持本提案实验设计的主要方法。