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.

项目概要 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 最近的工作有可能为哮喘和其他特应性疾病提供新的有效疗法。 证明 2 型辅助 T (Th2) 组织驻留记忆 T 细胞 (Trm) 持续存在于气道周围 过敏致敏后，它们迅速激活并驱动哮喘表型 限制 Th2-Trm 促炎功能的免疫调节机制。 该提案的目的是确定影响过敏原的因素。 肺部特定常驻 (r) Treg 的维持及其在再次暴露于 Th2-Trm 后的抑制 我们的中心假设是，过敏原特异性 rTreg 处于一个独特的位置。 从机制上讲，rTregs 可以有效抑制过敏性炎症。 使用基于 CXCR6 和 CCR8 趋化因子系统的 Th2-Trm 进行本地化 本提案将探讨这些。 使用新颖的实验系统来定义过敏性小鼠模型中 rTreg 的功能的问题 哮喘，包括屋尘螨 (HDM) 特异性细胞的多克隆过继转移、联体共生、单细胞 通过目标 1，我们将构建 HDM 特异性 T 细胞的 RNA 测序分析和 CRISPR-Cas9 基因编辑。 根据初步数据确定过敏原特异性 rTreg 在抑制肺部 Th2-Trm 中的作用。 表征 Tregs 的定位、持久性、抑制效力和转录特征 在目标 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。 组织住院医师计划并提供基因组编辑和系统生物学方面的额外培训，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 尼尔森博士将在波士顿儿童医院赞助的 K08 提案中执行这项工作， 马萨诸塞州总医院 (MGH) 免疫学和炎症疾病中心 (CIID) 在 Andrew Luster 博士的指导下，Nelson 博士制定了职业发展计划，其中包括： 高级显微镜、生物信息学和转化研究课程，以及指导实践 尖端研究方法的培训，以探索他的研究目标。K08 奖项的目标是 为纳尔逊博士提供成为独立的、由 NIH 资助的医生所需的技能和知识 具有特应性疾病人类疾病模型和非淋巴免疫调节方面专业知识的研究者 组织。