Lung resident Treg suppression of Th2 resident memory T cells in allergic asthma

过敏性哮喘中肺常驻 Treg 对 Th2 常驻记忆 T 细胞的抑制

• 批准号: 10664599
• 负责人: Ryan William Nelson
• 金额: $ 19.48万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-09 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10664599
• 关键词: Adoptive Transfer; Advisory Committees; Affect; Allergens; Allergic; Allergic Disease; Allergic inflammation; Asthma; Award; Bioinformatics; Biological Response Modifiers; Boston; CCL1 gene; CCL8 gene; CCR8 gene; CRISPR screen; CRISPR/Cas technology; CXCR6 gene; Candidate Disease Gene; Cell Maintenance; Cell Separation; Cells; Cellular biology; Chronic; Clustered Regularly Interspaced Short Palindromic Repeats; Data; Development Plans; Disease; Disease model; Exposure to; Extrinsic asthma; FOXP3 gene; Funding; Gene Expression Profile; General Hospitals; Genes; Genetic Transcription; Goals; Homeostasis; Immunology; Inflammation; Inflammatory; Inhalation; Interleukin-13; Kinetics; Knowledge; Ligands; Lung; Maintenance; Maps; Massachusetts; Mediating; Memory; Mentors; Mentorship; Metaplasia; Microscopy; Modeling; Mucous body substance; Mus; Parabiosis; Pediatric Hospitals; Peptides; Phase; Phenotype; Positioning Attribute; Pyroglyphidae; Regulatory T-Lymphocyte; Reporter; Research; Research Methodology; Research Personnel; Residencies; Resolution; Role; Structure of parenchyma of lung; System; Systems Biology; T cell differentiation; T-Lymphocyte; Testing; Th2 Cells; Therapeutic; Tissues; Training; Transgenic Organisms; Translational Research; United States National Institutes of Health; Work; airborne allergen; airway hyperresponsiveness; asthma model; career development; chemokine; chemokine receptor; effective therapy; eosinophil; genome editing; human disease; immunoregulation; in vivo; mouse model; mucosal site; novel; novel therapeutics; programs; recruit; response; single-cell RNA sequencing; skills; tissue resident memory T cell; transcriptomics

PROJECT SUMMARY Defining how FoxP3+ regulatory T cells (Tregs) limit amnestic responses to common allergens has the potential to provide new and effective therapies for asthma and other atopic diseases. Recent work has demonstrated that T helper type 2 (Th2) tissue-resident memory T cells (Trm) persist around the airways following allergic sensitization where they become rapidly activated and drive the asthma phenotype upon allergen re-exposure. The immunoregulatory mechanisms that limit the proinflammatory functions of Th2-Trm at mucosal sites are less clear. The objective of this proposal is to define the factors influencing allergen- specific resident (r) Treg maintenance in the lungs and their suppression of Th2-Trm following re-exposure to inhaled allergen. Our central hypothesis is that allergen-specific rTregs are positioned in a unique niche in the lung that enables potent suppression of allergic inflammation. Mechanistically, we hypothesize that rTregs co- localize with Th2-Trm based upon the CXCR6 and CCR8 chemokine systems. This proposal will explore these questions using novel experimental systems to define the function of rTregs in a mouse model of allergic asthma, including polyclonal adoptive transfer of house dust mite (HDM)-specific cells, parabiosis, single cell RNA sequencing analysis of HDM-specific T cells, and CRISPR-Cas9 gene editing. With Aim 1, we will build upon preliminary data to define the role of allergen-specific rTregs in suppressing lung Th2-Trm. We will characterize the localization, persistence, suppressive potency, and transcriptional profile of the Tregs that persist in the lung following allergen clearance. In Aim 2, we will determine the role of the CXCR6 and CCR8 chemokine systems in rTreg positioning and function. To identify additional factors important for the rTreg tissue residency program and provide additional training in genome editing and systems biology, we will perform an in vivo CRISPR-Cas9 screen on HDM-specific TCR transgenic cells isolated from the HDM memory lung. Dr. Nelson will perform the work in this K08 proposal, sponsored by Boston Children’s Hospital, at the Center for Immunology and Inflammatory diseases (CIID) at Massachusetts General Hospital (MGH) under the mentorship of Dr. Andrew Luster. Dr. Nelson has developed a career development plan consisting of coursework in advanced microscopy, bioinformatics, and translational research, along with mentored hands-on training in cutting-edge research methods to explore his research aims. The goal of this K08 award is to provide Dr. Nelson with the necessary skills and knowledge to become an independent, NIH-funded investigator with expertise in human disease models of atopic diseases and immunoregulation in non-lymphoid tissues.

项目摘要 定义FOXP3+调节性T细胞（Tregs）如何限制对常见过敏原的芳香反应 为哮喘和其他特应性疾病提供新的有效疗法的潜力。最近的工作有 证明T辅助型2型2（Th2）组织居住的记忆T细胞（TRM）在气道周围持续存在 遵循过敏敏感性，它们迅速激活并驱动哮喘表型 过敏原再暴露。限制TH2-TRM促炎功能的免疫调节机制 在粘膜位置不太清楚。该提议的目的是定义影响过敏原的因素 肺部的特定居民（R）TREG维持及其在重新暴露后对Th2-Trm的抑制 吸入过敏原。我们的中心假设是，过敏原特异性rtreg位于独特的位置 肺部可以潜在抑制过敏性注射。从机械上讲，我们假设Rtregs共同 基于CXCR6和CCR8趋化因子系统的Th2-TRM本地化。该建议将探索这些 使用新颖的实验系统来定义rtreg在过敏性小鼠模型中的功能的问题 哮喘，包括家用尘螨（HDM）特异性细胞的多克隆自适应转移，抛物线，单细胞 HDM特异性T细胞和CRISPR-Cas9基因编辑的RNA测序分析。使用AIM 1，我们将建立 根据初步数据来定义过敏原特异性rtreg在抑制肺Th2-Trm中的作用。我们将 表征Tregs的本地化，持久性，抑制效力和转录概况 在过敏原间隙后持续存在肺部。在AIM 2中，我们将确定CXCR6和CCR8的作用 RTREG定位和功能中的趋化因子系统。确定对Rtreg重要的其他因素 组织居住计划并提供基因组编辑和系统生物学的其他培训，我们将 在HDM分离的HDM特异性TCR转基因细胞上执行体内CRISPR-CAS9屏幕 内存肺。纳尔逊博士将在由波士顿儿童医院赞助的这项K08提案中执行这项工作 在马萨诸塞州综合医院（MGH）的免疫学和炎症性疾病中心（CIID） 在安德鲁·劳斯特（Andrew Luster）博士的心态下。尼尔森博士制定了一项职业发展计划，包括 高级显微镜，生物信息学和翻译研究的课程以及指导的动手 探索他的研究目的的尖端研究方法的培训。这个K08奖的目标是 向尼尔森博士提供必要的技能和知识，以成为独立的，NIH资助 研究人员具有人类疾病模型的专业知识和非淋巴管的免疫调节 组织。