TNFR2 Regulation of Leukocyte Recruitment in Glomerulonephritis

肾小球肾炎中白细胞募集的 TNFR2 调节

• 批准号: 8821615
• 负责人: Tanya N Mayadas
• 金额: $ 42.96万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8821615
• 关键词: Acute; Address; Affinity; Animal Model; Attenuated; Autocrine Communication; Behavior; Binding; Biological; Blocking Antibodies; Blood; Blood capillaries; Bone Marrow; CXCR3 gene; Cell Culture Techniques; Cells; Chemotactic Factors; Chimera organism; Chronic; Data; Development; Disease; Disease Progression; End stage renal failure; Endothelial Cells; Endothelium; Evolution; Functional disorder; Generations; Glomerular Capillary; Glomerular basement membrane antibody; Glomerulonephritis; Health; Human; IFNAR1 gene; Image; Imaging Techniques; Immunity; Immunotherapeutic agent; In Situ; Infiltration; Inflammation; Inflammatory; Inflammatory Response; Injection of therapeutic agent; Injury; Interferon Regulatory Factor 1; Interferon-alpha; Kidney; Kidney Diseases; Knockout Mice; Label; Lead; Leukocyte Trafficking; Leukocytes; Link; Membrane; Modeling; Molecular; Mononuclear; Mus; Nephritis; Organ; Organ Culture Techniques; Pathogenesis; Pathway interactions; Patients; Physiological; Production; Publishing; Receptor Signaling; Recombinants; Recruitment Activity; Regulation; Relative (related person); Reporter; Role; Route; Sampling; Signal Pathway; Signal Transduction; TNF gene; TNFRSF1A gene; TNFRSF1B gene; Testing; Time; Tissues; Transforming Growth Factor beta; Transgenic Mice; Vascular Endothelium; antibody conjugate; autocrine; base; capillary; chemokine; cytokine; glomerular basement membrane; glomerular endothelium; in vivo; insight; intravital microscopy; kidney cell; kidney imaging; macrophage; monocyte; neutrophil; new therapeutic target; novel; prevent; receptor; response; spatiotemporal; trafficking; transcription factor

DESCRIPTION (provided by applicant): Leukocyte accumulation is a hallmark of inflammatory renal diseases. TNF, a potent cytokine that regulates leukocyte trafficking is essential for the development of glomerulonephritis in animal models. Many of the pro- inflammatory responses of TNF in vivo can be attributed to its effects on the vascular endothelium and leukocyte influx. TNF relays its biological activities by two distinct receptors, TNFR1 and TNFR2 with most of TNF's effects on the endothelium being attributed to TNFR1. The contribution of TNFR2 is likely underestimated as it has a higher affinity for the membrane bound versus the soluble form of the cytokine and it is not constitutively expressed in all endothelial cell cultures. Our recently published data demonstrate that engagement of TNFR2 in cultured endothelial cells triggers Interferon regulatory factor-1 (IRF-1) induced IFNß synthesis, and subsequent autocrine signaling via the IFNα/ß receptor, to generate mononuclear cell chemoattractants. In vivo, acute renal inflammation induced by intravenously administered recombinant TNF results in neutrophil and monocyte recruitment in the kidney that requires TNFR1, while the TNFR2-IRF-1- IFNß autocrine loop is essential only for monocyte/macrophage accumulation. In a chronic model of anti- glomerular basement membrane nephritis, renal TNFR2 and IRF-1 but not TNFR1 are essential for sustained macrophage accumulation. Thus, our data identify a previously unrecognized TNFR2-IRF-1-IFNß-IFNα/ß receptor-signaling pathway in endothelial cells that promotes monocyte recruitment. These findings may have relevance in humans as TNFR2, absent in normal human kidney, is robustly induced on the glomerular endothelium of patients with anti-GBM nephritis. Our results raise two important questions: How does TNFR2 induce IRF-1, and what steps of leukocyte recruitment in the kidney are regulated by TNFR2 and its associated IFNß autocrine loop? These questions will be addressed in three specific aims: I) Delineate TNFR2 proximal intracellular signals that link TNFR2 to IRF-1 and IFNß production; II) Conduct intravital microscopy in the intact kidney to elucidate pathways of renal leukocyte recruitment triggered by TNF; and III) Elucidate the molecular requirements for renal TNFR2 and components of the IFNß autocrine loop in the dynamics of monocyte recruitment. For Aims II and III, we have developed a multiphoton intravital microscopy approach for time-lapse imaging of leukocyte behavior in the glomerular and peritubular capillaries of mice. Determining how TNF-induced amplification of IFNß expression and IFNß-signaling locally in the endothelium leads to monocyte accumulation in the kidney could result in important new insights into the pathogenesis of glomerulonephritis and lead to new therapeutic targets.

描述（由申请人提供）：白细胞积聚是炎症性肾病的标志。 TNF 是一种调节白细胞运输的强效细胞因子，对于动物模型中肾小球肾炎的发展至关重要。 TNF 体内的许多促炎反应可归因于其对血管内皮和白细胞流入的影响。 TNF 通过两种不同的受体 TNFR1 和 TNFR2 传递其生物活性，其中 TNF 对内皮细胞的大部分影响归因于 TNFR1。 TNFR2 的贡献可能被低估，因为与可溶形式的细胞因子相比，它对膜结合具有更高的亲和力，并且并非在所有内皮细胞培养物中组成型表达。我们最近发表的数据表明，TNFR2 在培养的内皮细胞中的参与会触发干扰素调节因子-1 (IRF-1) 诱导 IFNβ 合成，以及随后通过 IFNα/β 受体的自分泌信号传导，从而产生单核细胞趋化剂。在体内，静脉注射重组 TNF 诱导的急性肾脏炎症会导致中性粒细胞和单核细胞在肾脏中募集，这需要 TNFR1，而 TNFR2-IRF-1-IFNβ 自分泌环仅对单核细胞/巨噬细胞积累至关重要。在抗肾小球基底膜肾炎的慢性模型中，肾TNFR2和IRF-1而非TNFR1对于巨噬细胞的持续积累至关重要。因此，我们的数据确定了内皮细胞中以前未被识别的 TNFR2-IRF-1-IFNß-IFNα/ß 受体信号通路，可促进单核细胞募集。这些发现可能与人类相关，因为正常人肾脏中不存在的 TNFR2 在抗 GBM 肾炎患者的肾小球内皮上被强烈诱导。我们的结果提出了两个重要问题：TNFR2 如何诱导 IRF-1，以及 TNFR2 及其相关的 IFNß 自分泌环调节肾脏中白细胞募集的哪些步骤？这些问题将通过三个具体目标得到解决： I) 描绘将 TNFR2 与 IRF-1 和 IFNß 产生联系起来的 TNFR2 近端细胞内信号； II) 在完整肾脏中进行活体显微镜检查，以阐明 TNF 触发的肾白细胞募集途径； III) 阐明单核细胞募集动态中肾 TNFR2 和 IFNβ 自分泌环成分的分子需求。对于目标 II 和 III，我们开发了一种多光子活体显微镜方法，用于对小鼠肾小球和肾小管周围毛细血管中的白细胞行为进行延时成像。确定 TNF 诱导的内皮细胞局部 IFNβ 表达和 IFNβ 信号传导放大如何导致单核细胞在肾脏中积聚，可能会为肾小球肾炎的发病机制带来重要的新见解，并产生新的治疗靶点。