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: 9296123
• 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/9296123
• 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; 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; Human; Immune; Immune Complex Glomerulonephritis; Immunosuppressive Agents; Impairment; In Vitro; Infection; Infiltration; Inflammation; Inflammation Mediators; Inflammatory; Injury; Interleukin-6; Kidney; Kidney Diseases; Kidney Failure; Knock-in; Knock-in Mouse; Knowledge; Lead; Lectin; 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; Recruitment Activity; Renal function; Renal glomerular disease; Risk; Role; Serum; Signal Transduction; Steroids; Stress; TNF gene; TNFRSF1A 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; experimental study; improved; in vivo; macrophage; mimetics; mitogen-activated protein kinase p38; mutant; nephrotoxicity; novel; novel therapeutic intervention; novel therapeutics; overexpression; peptide drug; podocyte; prevent; promoter; protective effect; protective efficacy; public health relevance; receptor; repaired; synaptopodin; targeted treatment; therapeutic target; Acute Glomerulonephritis; Adverse effects; Affect; Albuminuria; Anti-Bacterial Agents; Anti-Inflammatory Agents; Anti-inflammatory; Antibodies; Bacterial Infections; Binding; Biological Preservation; CCL2 gene; Cells; Collagen; 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; Human; Immune; Immune Complex Glomerulonephritis; Immunosuppressive Agents; Impairment; In Vitro; Infection; Infiltration; Inflammation; Inflammation Mediators; Inflammatory; Injury; Interleukin-6; Kidney; Kidney Diseases; Kidney Failure; Knock-in; Knock-in Mouse; Knowledge; Lead; Lectin; 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; Recruitment Activity; Renal function; Renal glomerular disease; Risk; Role; Serum; Signal Transduction; Steroids; Stress; TNF gene; TNFRSF1A 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; experimental study; improved; in vivo; macrophage; mimetics; mitogen-activated protein kinase p38; mutant; nephrotoxicity; novel; novel therapeutic intervention; novel therapeutics; overexpression; peptide drug; podocyte; prevent; promoter; protective effect; protective efficacy; public health relevance; receptor; repaired; 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（通过应用提供）：小鼠中肾毒性血清（NTS）诱导的肾炎（NTN）的早期阶段的特征是过度感染和肾小球屏障功能障碍，代表了人类急性肾小球肾炎（GN）的良好模型。肾小球内皮和上皮室代表了早期治疗干预的有吸引力的靶标，这是由于它们在白细胞，蛋白尿和进行性肾衰竭的募集中的关键作用。当前旨在治疗或预防疾病进展的疗法不足和有限，这强调了控制与GN相关的新型治疗方法的需求。在这种情况下，细胞因子TNF是理想的靶标，因为它与人类GN和鼠NTN均涉及。不幸的是，遇到了意外的副作用，试图将TNF与其受体结合以治疗GN的受体，包括增加细菌感染的风险。因此，需要专门针对TNF的DELETERIOS效果但完整的有益效果的疗法是可取的。我们的初步研究表明，尖端肽模仿了TNF的讲座状结构域，并且不会干扰配体与受体的结合，从而干扰了NTN中至少两种途径在足细胞和肾小球内皮细胞（GEC）emise中进口的两种途径。一方面，肽首当其为GEC中的p38促炎途径，对于促炎性介体的产生至关重要，对于诱导eNOS功能障碍和白细胞浸润。另一方面，TIP肽可以保护足细胞屏障功能，这可能是通过在NTN中受到保护的足细胞中诱导PGE2产生的方法。在NTN的早期阶段，肽首当其为TNF，IL-6和MCP-1的全身产生，减少巨噬细胞浸润，水肿形成，恢复肾小球滤过率和突触型蛋白表达并消除蛋白尿症。我们的核心假设是，通过抑制TNF诱导的GEC和Podocytes中促炎和屏障的干扰途径的激活，尖端肽可以保护肾脏免受NTN的影响，因此，淡淡的白细胞浸润和肾小球损伤也无需干扰抗甲基甲虫的动作。在我们的三个特定目标中，我们 将在体内概念验证NTN小鼠研究（AIMS 1和2）结合基于机械的体外细胞研究（AIM 3）。我们将使用独特的模型和试剂，例如表达无功能性讲座域的TNF突变体的敲入小鼠（AIM 2）。此外，我们将在比较抗抗体耦合与裸尖肽（AIM 1A）的有效性时评估肽的局部与系统作用（AIM 1A），并在肾脏局部感染模型中测试肽，该模型由跨膜型TNF的内皮过表达（AIM 1B）启动。了解针对自身免疫性GN的尖端肽介导的保护作用的机制将提高我们对与肾病相关的炎症和抗炎症途径的了解，并将有助于进行临床试验以确定新型治疗性肽对其他免疫疾病的有效性。