Macrophages in human autoimmune tissue pathology

巨噬细胞在人类自身免疫组织病理学中的作用

基本信息

批准号：
10615897
负责人：
Laura T. Donlin
金额：
$ 44.5万
依托单位：
HOSPITAL FOR SPECIAL SURGERY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-04 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10615897
关键词：
Adult Affect American Architecture Arthralgia Autoimmune Autoimmune Diseases Automobile Driving Biological Assay Biological Models Blocking Antibodies CD8-Positive T-Lymphocytes Cartilage Cell Culture Techniques Chronic Chronic Disease Clinical Coculture Techniques Complex DTR gene Data Development Dinoprostone Disease EGF gene Epidermal Growth Factor Receptor Epiregulin Exhibits Experimental Models Feedback Fibroblasts Functional disorder Generations Human Hypoxia Immune Inflammation Inflammatory Interferon Type II Interferons Invaded Joints Knowledge Ligands Lymphocytic Infiltrate Macrophage Mediator Methods Molecular Pathogenesis Pathologic Pathology Pathway interactions Patients Pharmaceutical Preparations Phenotype Population Printing Prostaglandins Publications Reagent Reporting Rheumatoid Arthritis Shapes Small Interfering RNA Sorting Synovial Cell Synovial Fluid Synovial Membrane Synovial joint Synovitis System Testing Thromboplastin Tissues Translating angiogenesis bone cell type cytokine human disease improved inhibitor inhibitor therapy innovation intercellular communication joint inflammation knock-down novel novel therapeutics prevent response secretase single-cell RNA sequencing therapy development

项目摘要

In the autoimmune condition rheumatoid arthritis (RA), chronic inflammation reshapes cellular interactions and tissue architecture in patient joints. RA synovium is marked by expanded macrophage and fibroblast populations, extensive lymphocytic infiltration, angiogenesis and, ultimately, outgrowth beyond the natural tissue borders into cartilage and bone. In independent single-cell RNA-sequencing studies, we recently identified a novel macrophage phenotype found enriched in the synovium of RA patients (Zhang et al. Nat Immunol. 2019; Stephenson et al. Nat Comm. 2018). These macrophages express high levels of the EGF receptor (EGFR) ligand HB-EGF and are hereafter referred to as ‘HBEGF+ macrophages’. Our preliminary data also demonstrated that HBEGF+ macrophages are shaped by resident fibroblast factors along with pro-inflammatory cytokines including TNF (Kuo et al. Sci Transl Med. 2019). This newly activated macrophage state then feedbacks to activate fibroblast EGFR. Our central hypothesis posits that in RA synovial tissue, HBEGF+ macrophages are polarized by fibroblasts and in turn stimulate fibroblast pathologic activity. A critical prediction is that inhibition of mediators of this disease-associated crosstalk pathway will prevent tissue remodeling. We have developed an experimental cell culture model system to study HBEGF+ macrophage differentiation and the pathologic impact of their intercellular communication with synovial fibroblasts. We have also established a patient tissue ex vivo drug response assay, which has proven effective in defining how medications function in the complex cellular interactions of inflamed synovial tissue from clinically well-defined patients. Prior reports have established the relevance of macrophage-fibroblast crosstalk as a powerful regulator of RA pathology, but these reports lacked knowledge of the precise phenotypes of the synovial macrophages in RA (Rigor). With this new information and all methods and materials in place, we can look to define which of the known pathologic tissue factors induce this macrophage phenotype, specifically testing if IFNγ from abundant CD8+ T cells in the RA synovium, in combination with TNF and PGE2, induces the HBEGF+ macrophage phenotype (Aim 1). Furthermore, it is feasible to define how these macrophages drive RA synovial pathogenesis, in particular whether HB-EGF and/or epiregulin (EREG), a second EGF ligand expressed by HBEGF+ macrophages, drive both a hypoxic response and invasiveness in synovial fibroblasts and define which subtype of human synovial fibroblasts exhibit invasiveness in response to these EGF ligands (Aim 2). Finally, we are able to test the impact of potential inhibitors on HBEGF+ macrophage generation and pathologic function, focusing on the ADAM17/iRhom2 complex, which controls both the release of HB-EGF and TNF from macrophages (Aim 3). Completion of these aims will leverage our finding of human disease-associated macrophages, define mechanisms of a new pathway driving crosstalk between synovial macrophages and fibroblasts (Innovation), and lay the founding for our long- term objective of translating molecular findings to develop new therapies for the substantial number of RA patients not responding currently to therapies (Significance).

在类风湿性关节炎 (RA) 自身免疫性疾病中，慢性炎症会重塑细胞相互作用并 RA 滑膜的组织结构以巨噬细胞和成纤维细胞群增多为特征，广泛的淋巴细胞浸润、血管生成，并最终生长到自然组织边界之外在独立的单细胞 RNA 测序研究中，我们最近发现了一种新的软骨和骨骼。在 RA 患者的滑膜中发现巨噬细胞表型丰富（Zhang et al. Nat Immunol. 2019； Stephenson 等人，2018）。这些巨噬细胞表达高水平的 EGF 受体 (EGFR)。我们的初步数据也证明了配体 HB-EGF，以下称为“HBEGF+巨噬细胞”。 HBEGF+巨噬细胞是由常驻成纤维细胞因子和促炎细胞因子形成的包括 TNF（Kuo et al. Sci Transl Med. 2019）。这种新激活的巨噬细胞状态随后反馈到我们的中心假设认为，在 RA 滑膜组织中，HBEGF+ 巨噬细胞是被成纤维细胞极化并反过来刺激成纤维细胞病理活动。一个关键的预测是抑制。这种与疾病相关的串扰途径的介质将阻止组织重塑。研究 HBEGF+ 巨噬细胞分化及其病理影响的实验细胞培养模型系统我们还建立了离体患者组织。药物反应测定，已被证明可以有效地确定药物在复杂细胞中的作用先前的报告已经确定了临床上明确的患者的发炎滑膜组织之间的相互作用。巨噬细胞-成纤维细胞串扰作为 RA 病理学的强大调节因子的相关性，但这些报告缺乏了解 RA 滑膜巨噬细胞的精确表型（Rigor）。所有方法和材料都到位后，我们可以确定哪些已知的病理组织因素会诱发这种巨噬细胞表型，专门测试 RA 滑膜中丰富的 CD8+ T 细胞是否存在 IFNγ，与 TNF 和 PGE2 结合，诱导 HBEGF+ 巨噬细胞表型（目标 1）。可以确定这些巨噬细胞如何驱动 RA 滑膜发病机制，特别是 HB-EGF 和/或上皮调节蛋白 (EREG) 是 HBEGF+ 巨噬细胞表达的第二种 EGF 配体，可驱动缺氧反应滑膜成纤维细胞的侵袭性和侵袭性，并确定人类滑膜成纤维细胞的亚型对这些 EGF 配体的侵袭性反应（目标 2）最后，我们能够测试潜在的影响。 HBEGF+巨噬细胞生成和病理功能的抑制剂，重点关注 ADAM17/iRhom2 复合物，控制巨噬细胞释放 HB-EGF 和 TNF（目标 3）。目标将利用我们对人类疾病相关巨噬细胞的发现，定义新途径的机制驱动滑膜巨噬细胞和成纤维细胞之间的串扰（创新），并为我们的长期转化分子发现为大量 RA 开发新疗法的长期目标目前对治疗没有反应的患者（意义）。