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

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

项目概要/摘要该提案是一项为期五年的研究和培训计划，科学重点是 CD8 T 之间的相互作用我们发现类风湿性关节炎（RA）患者滑膜组织中的细胞和成纤维细胞。 RA 滑膜中的大多数 CD8 T 细胞具有不寻常的表型，其特征是经典的低表达相反，这些细胞表达大量的细胞毒性蛋白，例如颗粒酶 B、穿孔素和颗粒溶素。颗粒酶 K，诱导滑膜成纤维细胞产生 IL-6 等促炎因子。根据我们的初步数据，我们相信颗粒酶 K 的作用不仅仅是激活滑膜成纤维细胞。认为颗粒酶 K 激活补体系统以诱导所谓的滑膜炎症启动成纤维细胞，其特征是代谢重编程和对刺激的增强激活。这里提出的具体目标将研究 CD8 T 细胞和滑膜成纤维细胞在三个方面的相互作用目标 1 询问人类颗粒酶 K 是否诱导代谢重编程。与炎症引发相关以及补体是否介导颗粒酶的下游效应 K. Aim 2 使用成像质谱流式细胞术、传统免疫荧光和空间转录组学来评估颗粒酶 K+ CD8 T 细胞及其与成纤维细胞亚群和补体沉积的关系 Aim 3 使用炎症性关节炎小鼠模型来确定。颗粒酶 K 对炎症性关节炎临床和细胞指标的影响。这项研究结合了机制研究、患者来源的样本、小鼠模型和尖端技术转录组技术为候选人提供翻译几个关键方面的新培训候选人的直接职业发展目标是获得显微镜方面的经验。技术、空间转录组学、细胞代谢、自身免疫性疾病的小鼠模型，以及候选人和她的导师都描述了具体的职业发展计划。迈克尔·布伦纳 (Michael Brenner) 医学博士是淋巴细胞生物学和滑膜炎症方面的专家。候选人将在其技能提升领域获得专家咨询委员会的支持。长期目标是获得终身教授职位，从事整合高维度研究的研究通过详细的机制研究分析患者样本，以表征组织内的细胞相互作用在风湿病领域，最终目标是确定新的细胞和分子候选药物自身免疫性疾病的诊断和治疗。