Architecture and interactions of Granzyme K+ CD8 T cells in inflamed synovium in rheumatoid arthritis

类风湿关节炎炎症滑膜中颗粒酶 K CD8 T 细胞的结构和相互作用

• 批准号: 10506508
• 负责人: Anna Helena Jonsson
• 金额: $ 17.39万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2023-06-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10506508
• 关键词: Advisory Committees; Animal Model; Architecture; Area; Autoimmune Diseases; Bioinformatics; Biological; CCL2 gene; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; Cells; Clinical; Collagen Arthritis; Complement; Complement Activation; Computational Biology; Core Facility; Crohn' s disease; Cytometry; Data; Deposition; Development Plans; Diagnosis; Disease; Doctor of Philosophy; Exhibits; FRAP1 gene; Faculty; Fibroblasts; Flow Cytometry; Frequencies; Goals; Granzyme; Histology; Human; Image; Immunofluorescence Immunologic; Immunology; In Situ; Inflammation; Inflammatory; Inflammatory Arthritis; Interleukin-6; Irrigation; Kidney; Laboratories; Link; Liquid substance; Lung; Lupus Nephritis; Lymphocyte; Measures; Mediating; Mediator of activation protein; Mentors; Mentorship; Metabolic; Metabolism; Microscopy; Modeling; Molecular; Mus; Pathogenesis; Pathway interactions; Patients; Phenotype; Play; Positioning Attribute; Production; Proteins; Reactive Oxygen Species; Reporting; Research; Research Personnel; Research Training; Reverse Transcriptase Polymerase Chain Reaction; Rheumatism; Rheumatoid Arthritis; Rheumatology; Role; SARS-CoV-2 infection; Sampling; Stimulus; Synovial Fluid; Synovial Membrane; Synovitis; T-Lymphocyte; T-Lymphocyte Subsets; TNF gene; Techniques; Technology; Time; Tissue Sample; Tissue imaging; Tissues; Training; Ulcerative Colitis; Wild Type Mouse; base; biobank; career; career development; complement system; cytotoxic; dimensional analysis; experience; experimental study; genetic signature; granulysin; gut inflammation; high dimensionality; improved; in vivo; inflammatory marker; instructor; metabolomics; mouse model; perforin; severe COVID-19; single-cell RNA sequencing; skills; tenure track; transcriptome sequencing; transcriptomics

Project Summary/Abstract This proposal is a five-year research and training plan with a scientific focus on interactions between CD8 T cells and fibroblasts in synovial tissue from patients with rheumatoid arthritis (RA). We have found that the majority of CD8 T cells in RA synovium have an unusual phenotype characterized by low expression of classic cytotoxic proteins such as granzyme B, perforin, and granulysin. Instead, these cells express high amounts of granzyme K, which induces synovial fibroblasts to produce pro-inflammatory factors such as IL-6. Based on our preliminary data, we believe that granzyme K does more than simply activate synovial fibroblasts. We believe that granzyme K activates the complement system to induce so-called inflammatory priming of synovial fibroblasts, characterized by metabolic reprogramming and augmented activation to stimuli. The specific aims proposed here will investigate the interactions of CD8 T cells and synovial fibroblasts in three complementary ways. Aim 1 interrogates whether human granzyme K induces the metabolic reprograming associated with inflammatory priming and whether complement mediates the downstream effects of granzyme K. Aim 2 uses imaging mass cytometry, traditional immunofluorescence, and spatial transcriptomics to evaluate granzyme K+ CD8 T cells and their relationship with fibroblast subsets and complement deposition within the context of human synovial tissues. Aim 3 uses mouse models of inflammatory arthritis to determine the effects of granzyme K on clinical and cellular measures in inflammatory arthritis. This study combines mechanistic studies, patient-derived samples, mouse models, and cutting-edge transcriptomic technologies to provide the candidate new training in several key aspects of translational immunology. The candidates immediate career development goals are to gain experience with microscopy techniques, spatial transcriptomics, cellular metabolism, mouse models of autoimmune diseases, and bioinformatic analysis. A specific career development plan is described by both the candidate and her mentor Dr. Michael Brenner, MD, an expert in lymphocyte biologic and synovial inflammation. She also has the support of an Advisory Committee of experts in the areas in which she will build her skills. The candidate’s long-term goal is to attain a tenure-track faculty position pursuing research that integrates high-dimensional analysis of patient samples with detailed mechanistic studies to characterize cellular interactions within tissues in rheumatologic diseases, with the ultimate goal of identifying new cellular and molecular candidates for diagnosis and treatment of autoimmune diseases.

项目摘要/摘要 该提案是一项为期五年的研究和培训计划，科学的重点是CD8 T之间的相互作用 类风湿关节炎患者的滑膜组织中的细胞和成纤维细胞（RA）。我们发现 RA滑膜中的大多数CD8 T细胞具有不寻常的表型，其特征是经典的表达低。 细胞毒性蛋白，例如颗粒酶B，穿孔蛋白和颗粒素。相反，这些细胞表达了大量的 Granzyme K，诱导滑膜成纤维细胞产生促炎性因子，例如IL-6。基于 我们的初步数据，我们认为Granzyme K不仅可以简单地激活滑膜成纤维细胞。我们 相信颗粒酶K激活完成系统以诱导滑膜的炎症启动 成纤维细胞，其特征是代谢重编程和增强激活刺激。 此处提出的具体目的将研究CD8 T细胞和滑膜成纤维细胞的相互作用三个 完全方式。 AIM 1询问人类颗粒酶是否诱导代谢重编程 与炎症性启动以及补体是否介导颗粒的下游作用有关 K. AIM 2使用成像质量细胞术，传统的免疫荧光和空间转录组学 评估颗粒酶K+ CD8 T细胞及其与成纤维细胞子集的关系和完成沉积 在人类滑膜组织的背景下。 AIM 3使用炎症性关节炎的小鼠模型来确定 颗粒酶K对炎症性关节炎的临床和细胞测量的影响。 这项研究结合了机械研究，患者衍生的样品，小鼠模型和尖端 转录组技术在翻译的几个关键方面提供候选新培训 免疫学。候选人的立即职业发展目标是获得显微镜的经验 技术，空间转录组学，细胞代谢，自身免疫性疾病的小鼠模型和 生物信息学分析。候选人和她的精神都描述了一项特定的职业发展计划 迈克尔·布伦纳（Michael Brenner）博士，医学博士，淋巴细胞生物学和滑膜注射专家。她也有 支持她将要建立技能的领域的专家咨询委员会的支持。候选人的 长期目标是达到终身教师职位，以融入高维度的研究 分析具有详细机械研究的患者样品，以表征组织内的细胞相互作用 在风湿病疾病中，最终目的是确定新的细胞和分子候选者 自身免疫性疾病的诊断和治疗。