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: 8963148
• 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/8963148
• 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; Human; Immune; Immune Complex Glomerulonephritis; In Vitro; Infection; Infiltration; Inflammation; Inflammation Mediators; Inflammatory; Injury; Interleukin-6; Kidney; Kidney Diseases; Kidney Failure; 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; podocyte; prevent; promoter; protective effect; protective efficacy; public health relevance; 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（由申请方提供）：小鼠肾毒性血清（NTS）诱导的肾炎（NTN）的早期特征为过度炎症和肾小球屏障功能障碍，是人类急性肾小球肾炎（GN）的良好模型。由于肾小球内皮和上皮细胞在白细胞募集、蛋白尿和进行性肾衰竭中的关键作用，它们代表了早期治疗干预的有吸引力的靶点。目前旨在治疗或预防疾病进展的疗法是不足和有限的，强调需要新的治疗方法来控制与GN相关的炎症。在这种情况下，细胞因子TNF是一个理想的目标，因为它是关键参与人类GN和鼠NTN。不幸的是，试图钝化TNF与其受体的结合以治疗GN遇到了意想不到的副作用，包括细菌感染的风险增加。因此，期望特异性靶向TNF的抗肿瘤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，它模仿TNF的凝集素样结构域，并不干扰配体的结合，其受体，干扰至少两个途径importat足细胞和肾小球内皮细胞（GEC）死亡在NTN。一方面，该肽钝化GEC中的p38促炎途径，这对于促炎介质的产生、eNOS功能障碍的诱导和白细胞浸润是重要的。另一方面，TIP肽保护足细胞屏障功能，可能是指通过诱导足细胞中PGE 2的产生，在NTN中显示出保护性。在NTN的早期阶段，该肽减弱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足蛋白表达，并消除白蛋白尿。我们的中心假设是TIP肽通过分别抑制GEC和足细胞中TNF诱导的促炎和屏障破坏性途径的激活来保护肾脏免受NTN的影响，因此钝化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， 将结合联合收割机体内概念验证NTN小鼠研究（目标1和2）、基于足细胞和GEC的机制体外细胞研究（目标3）。我们将使用独特的模型和试剂，例如表达TNF突变体而没有功能性凝集素样结构域的敲入小鼠（目的2）。此外，我们将通过比较抗-β 3（IV）抗体偶联的TIP肽与裸TIP肽的功效（目的1A）并在由跨膜TNF的内皮过表达引发的肾局部炎症模型中测试肽（目的1B）来评估肽的局部与全身作用。了解TIP肽介导的针对自身免疫性GN的保护作用的机制将推进我们对与肾病相关的炎症和抗炎途径的认识，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性、炎症性疾病的功效。