Novel peptide-based strategy to inhibit deleterious TNF signaling in nephrotoxic nephritis

基于肽的新型策略抑制肾毒性肾炎中有害的 TNF 信号传导

• 批准号: 9115574
• 负责人: Rudolf Lucas
• 金额: $ 22.8万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9115574
• 关键词: Acute Glomerulonephritis; Adverse effects; Affect; Albuminuria; Anti-Bacterial Agents; Anti-Inflammatory Agents; Anti-inflammatory; Antibodies; Bacterial Infections; Binding; Biological Preservation; CCL2 gene; Cells; Collagen Type IV; Complex; Conduct Clinical Trials; Coupled; Coupling; Development; Dinoprostone; Disease; Disease Progression; Edema; Endothelial Cells; Epithelial; Epitopes; Etiology; Functional disorder; Generations; Glomerular Filtration Rate; Glomerulonephritis; Goals; Health; Human; Immune; Immune Complex Glomerulonephritis; In Vitro; Infection; Infiltration; Inflammation; Inflammation Mediators; Inflammatory; Injury; Interleukin-6; Kidney; Kidney Diseases; Kidney Failure; Knock-in; Knock-in Mouse; Knowledge; Lead; Lectin; Left; Leukocytes; Ligand Binding; Link; MAPK14 gene; Mediating; Mediator of activation protein; Methods; Modeling; Mus; Nephritis; Nephrotoxic; Onset of illness; Outcome; Participant; Pathogenesis; Pathway interactions; Patients; Peptides; Pharmaceutical Preparations; Phase; Physiological; Play; Predisposition; Property; Proteinuria; Reagent; Renal function; Renal glomerular disease; Risk; Role; Serum; Signal Transduction; Staging; Steroids; Stress; TNF gene; Testing; Therapeutic; Therapeutic Intervention; Time; Transgenic Mice; Tumor Necrosis Factor Receptor; Western Blotting; Wild Type Mouse; base; caveolin 1; chemokine; comparative efficacy; conventional therapy; cytokine; improved; in vivo; macrophage; meetings; mimetics; mitogen-activated protein kinase p38; mutant; novel; novel therapeutic intervention; novel therapeutics; overexpression; peptide drug; podocyte; prevent; promoter; protective effect; protective efficacy; receptor; repaired; research study; synaptopodin; targeted treatment; therapeutic target

 DESCRIPTION (provided by applicant): The early stage of nephrotoxic serum (NTS)-induced nephritis (NTN) in mice is characterized by excessive inflammation and glomerular barrier dysfunction and represents a good model for human acute glomerulonephritis (GN). Glomerular endothelial and epithelial compartments represent attractive targets for early therapeutic intervention, due to their crucial roles in the recruitment of leukocytes, proteinuria and progressive renal failure. Current therapies aiming to treat or prevent progression of the disease are insufficient and limited, stressing the need for novel therapeutic approaches that control inflammation associated with GN. In this context, the cytokine TNF is an ideal target, since it is critically involved in both human GN and in murine NTN. Unfortunately, attempts to blunt TNF binding to its receptors for the treatment of GN were met with unexpected side effects, including increased risk for bacterial infection. Therefore, therapies that specifically target the deleterios effects of TNF, but leave its beneficial effects intact are desirable. Our preliminary studies indicate that the TIP peptide, which mimics TNF's lectin-like domain and which does not interfere with the ligand's binding to its receptors, interferes with at least two pathways importat in podocyte and glomerular endothelial cells (GEC) demise in NTN. On the one hand, the peptide blunts the p38 pro-inflammatory pathway in GEC, important for the generation of pro-inflammatory mediators, for the induction of eNOS dysfunction and for leukocyte infiltration. On the other hand, TIP peptide protects podocyte barrier function, possibly means by inducing PGE2 generation in podocytes, shown to be protective in NTN. In the early phase of NTN, the peptide blunts systemic generation of TNF, IL-6 and MCP-1, reduces macrophage infiltration, edema formation, restores glomerular filtration rate and synaptopodin expression and abrogates albuminuria. Our central hypothesis is that the TIP peptide protects the kidney against NTN by suppressing TNF-induced activation of pro-inflammatory and barrier-disruptive pathways in GEC and podocytes, respectively, as such blunting leukocyte infiltration and glomerular injury, without interfering with the anti-bacterial actions of the cytokine. In our three specific aims, we will combine in vivo proof-of-concept NTN mouse studies (aims 1 and 2) podocyte- and GEC-based with mechanistic in vitro cell studies (aim 3). We will use unique models and reagents, such as knock-in mice expressing a TNF mutant without functional lectin-like domain (aim 2). Moreover, we will assess local vs. systemic effects of the peptide upon comparing the efficacy of anti-3(IV) antibody-coupled vs. naked TIP peptide (aim 1A) and testing the peptide in a renal local inflammation model, initiated by endothelial overexpression of transmembrane TNF (aim 1B). Understanding the mechanism of TIP peptide- mediated protective effects against auto-immune GN will advance our knowledge on the inflammatory and anti- inflammatory pathways relevant for nephropathy and will help to conduct clinical trials to determine the efficacy of novel therapeutic peptides against other immune, inflammatory diseases.

 描述（由申请人提供）：小鼠肾毒性血清（NTS）诱导的肾炎（NTN）的早期阶段以过度炎症和肾小球屏障功能障碍为特征，代表了人类急性肾小球肾炎（GN）的良好模型。肾小球内皮和上皮区室是早期治疗干预的有吸引力的目标，因为它们在白细胞募集、蛋白尿和进行性肾衰竭中发挥着关键作用。目前旨在治疗或预防该疾病进展的疗法不足且有限，这强调需要控制与 GN 相关炎症的新治疗方法。在这种情况下，细胞因子 TNF 是一个理想的靶标，因为它与人类 GN 和小鼠 NTN 密切相关。不幸的是，通过削弱 TNF 与其受体的结合来治疗 GN 的尝试遇到了意想不到的副作用，包括增加细菌感染的风险。因此，需要专门针对 TNF 的有害作用但保持其有益作用完整的疗法。我们的初步研究表明，TIP 肽模拟 TNF 的凝集素样结构域，并且不会干扰配体与其受体的结合，它会干扰 NTN 中足细胞和肾小球内皮细胞 (GEC) 死亡的至少两条重要途径。一方面，该肽会削弱 GEC 中的 p38 促炎途径，这对于促炎介质的产生、诱导 eNOS 功能障碍和白细胞浸润非常重要。另一方面，TIP 肽可保护足细胞屏障功能，可能是通过诱导足细胞中 PGE2 的生成来保护足细胞屏障功能，这在 NTN 中显示出保护作用。在 NTN 的早期阶段，该肽会减弱 TNF、IL-6 和 MCP-1 的全身生成，减少巨噬细胞浸润、水肿形成，恢复肾小球滤过率和突触足蛋白表达，并消除蛋白尿。我们的中心假设是，TIP 肽通过抑制 TNF 诱导的 GEC 和足细胞中促炎和屏障破坏途径的激活来保护肾脏免受 NTN 侵害，从而减弱白细胞浸润和肾小球损伤，而不干扰细胞因子的抗菌作用。在我们的三个具体目标中，我们 将把基于足细胞和 GEC 的体内概念验证 NTN 小鼠研究（目标 1 和 2）与机械体外细胞研究（目标 3）结合起来。我们将使用独特的模型和试剂，例如表达不具有功能性凝集素样结构域的 TNF 突变体的敲入小鼠（目标 2）。此外，我们将通过比较抗 α3(IV) 抗体偶联与裸露 TIP 肽（目标 1A）的功效，并在由跨膜 TNF 的内皮过度表达引发的肾脏局部炎症模型中测试该肽（目标 1B），来评估该肽的局部与全身效应。了解 TIP 肽介导的针对自身免疫性肾小球肾炎的保护作用的机制将增进我们对与肾病相关的炎症和抗炎途径的了解，并将有助于进行临床试验以确定新型治疗肽针对其他免疫、炎症疾病的功效。