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.

项目总结 定义FoxP3+调节性T细胞(Treg)如何限制对常见过敏原的遗忘反应 有可能为哮喘和其他特应性疾病提供新的有效治疗方法。最近的工作有 证明辅助性T细胞2型(Th2)组织驻留记忆T细胞(Trm)持续存在于呼吸道 在过敏性致敏后，它们迅速激活并驱动哮喘表型 过敏原再次暴露。限制Th2-Trm促炎功能的免疫调节机制 在粘膜部位则不太清楚。这项建议的目的是界定影响过敏原的因素- 特异居民(R)Treg在肺中的维持及其对再暴露后Th2-Trm的抑制 吸入过敏原。我们的中心假设是过敏原特异性rTregs位于 能够有效抑制过敏性炎症的肺。从机制上讲，我们假设rTregs共同- 根据CXCR6和CCR8趋化因子系统，用Th2-Trm进行本地化。本提案将探讨这些问题 使用新的实验系统确定rTregs在过敏性小鼠模型中的功能的问题 哮喘，包括多克隆过继转移屋尘螨(HDM)特异性细胞、异种共生、单细胞 HDM特异性T细胞的RNA测序分析和CRISPR-Cas9基因编辑。有了目标1，我们将建立 根据初步数据确定变应原特异性rTregs在抑制肺Th2-Trm中的作用。我们会 描述Tregs的本地化、持久性、抑制效力和转录图谱 过敏原清除后持续留在肺内。在目标2中，我们将确定CXCR6和CCR8的作用 趋化因子系统在rTreg中的定位和功能。确定对rTreg很重要的其他因素 组织驻院计划，并提供额外的基因组编辑和系统生物学培训，我们将 对从HDM分离的HDM特异性TCR转基因细胞进行体内CRISPR-Cas9筛选 记忆肺。纳尔逊博士将在这份由波士顿儿童医院赞助的K08提案中开展工作， 马萨诸塞州总医院(MGH)免疫和炎症性疾病中心(CIID) 在安德鲁·勒斯特博士的指导下。纳尔逊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方面的课程，以及有指导的实践 接受前沿研究方法培训，探索自己的研究目标。这个K08奖项的目标是 为Nelson博士提供必要的技能和知识，使其成为一名由NIH资助的独立的 在特应性疾病的人类疾病模型和非淋巴组织免疫调节方面具有专业知识的研究员 纸巾。