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

项目摘要/摘要 这项建议是一项为期五年的研究和培训计划，科学重点是CD8 T细胞之间的相互作用 类风湿关节炎(RA)滑膜组织中的细胞和成纤维细胞。我们发现， RA滑膜中大多数CD8T细胞具有一种不寻常的表型，其特征是Classic低表达 细胞毒性蛋白，如颗粒酶B、穿孔素和颗粒溶素。相反，这些细胞表达大量的 颗粒酶K，可诱导滑膜成纤维细胞产生IL-6等促炎因子。基于 我们的初步数据显示，我们认为颗粒酶K不只是简单地激活滑膜成纤维细胞。我们 相信颗粒酶K激活补体系统，诱导所谓的滑膜炎性启动 成纤维细胞，以代谢重新编程和对刺激的增强激活为特征。 这里提出的特定目标将研究CD8T细胞和滑膜成纤维细胞之间的相互作用。 互补的方式。目的1探讨人颗粒酶K是否诱导代谢重编程 与炎症启动相关以及补体是否介导颗粒酶的下游效应 K.Aim 2使用成像质量细胞术、传统免疫荧光和空间转录学来 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 在人类滑膜组织的背景下。AIM 3使用小鼠炎症性关节炎模型来确定 颗粒酶K对炎性关节炎临床和细胞指标的影响。 这项研究结合了机械学研究、患者衍生的样本、小鼠模型和尖端技术 转录技术，为应聘者提供翻译的几个关键方面的新培训 免疫学。应聘者近期的职业发展目标是获得显微镜方面的经验。 技术，空间转录学，细胞代谢，自身免疫性疾病的小鼠模型，以及 生物信息学分析。应聘者和她的导师都描述了具体的职业发展计划 迈克尔·布伦纳博士，医学博士，淋巴细胞生物学和滑膜炎症专家。她还拥有 在她将提高技能的领域得到专家咨询委员会的支持。候选人的 长期目标是获得终身教职，从事整合高维研究的研究 通过详细的机制研究对患者样本进行分析，以表征组织内的细胞相互作用 在风湿病方面，最终目标是确定新的细胞和分子候选 自身免疫性疾病的诊断和治疗。

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.