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

• 批准号: 10158419
• 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
• 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/10158419
• 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; Infiltration; Inflammation; Inflammatory; Interferon Type II; Interferons; Joints; Knowledge; Lead; Ligands; Lymphocyte; Mediator of activation protein; Methods; Molecular; Pathogenesis; Pathologic; Pathology; Pathology Report; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Population; Prostaglandins; Publications; Reagent; Reporting; Rheumatoid Arthritis; Shapes; Small Interfering RNA; Synovial Cell; Synovial Fluid; Synovial Membrane; Synovial joint; Synovitis; System; TNF gene; Testing; Therapeutic; Thromboplastin; Tissues; Translating; angiogenesis; bone; cell type; cytokine; human disease; improved; inhibitor/antagonist; innovation; intercellular communication; joint inflammation; knock-down; macrophage; novel; novel therapeutics; prevent; response; secretase; single-cell RNA sequencing; therapy development; 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; Infiltration; Inflammation; Inflammatory; Interferon Type II; Interferons; Joints; Knowledge; Lead; Ligands; Lymphocyte; Mediator of activation protein; Methods; Molecular; Pathogenesis; Pathologic; Pathology; Pathology Report; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Population; Prostaglandins; Publications; Reagent; Reporting; Rheumatoid Arthritis; Shapes; Small Interfering RNA; Synovial Cell; Synovial Fluid; Synovial Membrane; Synovial joint; Synovitis; System; TNF gene; Testing; Therapeutic; Thromboplastin; Tissues; Translating; angiogenesis; bone; cell type; cytokine; human disease; improved; inhibitor/antagonist; innovation; intercellular communication; joint inflammation; knock-down; macrophage; novel; novel therapeutics; prevent; response; secretase; single-cell RNA sequencing; therapy development

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).

在自身免疫性条件下，类风湿关节炎（RA），慢性炎症会改造细胞相互作用和 患者关节中的组织结构。 Ra Syronovium以扩大的巨噬细胞和成纤维细胞种群为特征， 广泛的淋巴细胞浸润，血管生成，最终将天然组织边界以外的生长 软骨和骨头。在独立的单细胞RNA测序研究中，我们最近确定了一种新颖的 发现巨噬细胞表型富含RA患者的滑膜（Zhang等，NatImmunol。2019; 斯蒂芬森等人。纳特通讯。 2018）。这些巨噬细胞表达高水平的EGF受体（EGFR） 配体HB-EGF，以下称为“ HBEGF+巨噬细胞”。我们的初步数据也证明了 HBEGF+巨噬细胞由居民成纤维细胞因子以及促炎性细胞因子塑造 包括TNF（Kuo等人Sci TransborMed。2019）。这种新激活的巨噬细胞随后反馈给 激活成纤维细胞EGFR。我们的中心假设认为，在RA滑膜组织中，HBEGFF+巨噬细胞是 由成纤维细胞偏振，进而刺激成纤维细胞病理活性。一个关键的预测是抑制 该疾病相关的串扰途径的介体将防止组织重塑。我们已经开发了 实验细胞培养模型系统研究HBEGF+巨噬细胞分化和病理影响 它们与滑膜成纤维细胞的细胞间通信。我们还建立了一个病人的组织 药物反应评估，已证明有效定义药物如何在复杂细胞中的功能 临床定义明确的患者发炎的滑膜组织的相互作用。先前的报告已经建立了 巨噬细胞 - 纤维细胞串扰的相关性是RA病理学的强大调节剂，但这些报告缺乏 了解RA（严格）中滑膜巨噬细胞的精确表型的知识。有了这些新信息， 所有方法和材料都可以定义哪些已知病理组织因素影响 这种巨噬细胞表型，专门测试是否来自RA滑膜中丰富的CD8+ T细胞的IFNγ，在 结合TNF和PGE2，诱导HBEGFF+巨噬细胞表型（AIM 1）。此外，是 可行的来定义这些巨噬细胞如何驱动RA滑膜发病机理，特别是HB-EGF和/或 Epiregulin（EREG）是由HBEGF+巨噬细胞表达的第二个EGF配体，这两种反应均可驱动于低氧反应 以及滑膜成纤维细胞中的侵袭性，并定义人类滑膜成纤维细胞的哪种亚型 响应这些EGF配体的侵入性（AIM 2）。最后，我们能够测试潜力的影响 HBEGF+巨噬细胞生成和病理功能的抑制剂，重点是ADAM17/IRHOM2 复合物，可以控制巨噬细胞中HB-EGF和TNF的释放（AIM 3）。这些完成 目的将利用我们发现与人类疾病相关的巨噬细胞的发现，定义新途径的机制 在滑膜巨噬细胞和成纤维细胞（创新）之间驱动串扰，并为我们的长期建立 转换分子发现以开发大量RA的新疗法的术语目标 患者目前未对疗法做出反应（显着性）。