Molecular Mechanisms of Peritoneal Fibrosis- Proteinkinase C alpha and Peritoneal Dialysis

腹膜纤维化的分子机制-蛋白激酶Cα与腹膜透析

• 批准号: 284208194
• 负责人: Professor Dr. Hermann Haller
• 金额: --
• 依托单位: Klinik für Nieren- und Hochdruckerkrankungen
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/284208194?language=en
• 关键词: Molecular; Mechanisms; Peritoneal; Fibrosis; Proteinkinase

In our previous experiments we were able to demonstrate in the experiments supported by that PKCα is the only classical PKC isoform expressed in MPMCs. PKCα protein expression and activation is upregulated in MPMCs in vitro under PD-simulating conditions and in vivo after prolonged exposure to PDF. Whereas in WT mice chronic PDF administration leads to severe morphologic, structural, and functional changes of the PM, the functional blockade of PKCα activity with Go6976 completely protected PM structure and function. PKCα deficiency showed a comparable benefit, suggesting a crucial role of PKCα in PM disturbances. Based on these findings in the last funding period we propose a novel therapeutic approach to prevent PD-fluid induced inflammation, neoangiogenesis and fibrosis. 1. Using an antisense strategy against PKC isoform alpha regularly applied with the PD fluid during dialysis we want to inhibit the glucose-induced inflammation, neoangiogenesis and fibrosis. The specific antisense approach prevents the side effects of other PKC inhibitors such as Gö, which may also affect multiple kinases from a variety of protein kinase families and usually have other pharmacological actions leading to different effects in vitro and in vivo independently from PKC inhibition. 2. We will analyze the therapeutic effect of our strategy on the different cell types of the peritoneum. We have evidence from our previous research that mesothelial cells are a main therapeutic target. However, we have also seen previously that PKC-a in endothelial cells, macrophages and fibroblasts plays an important role. We therefore plan to use single nucleus gene expression analysis to define our therapeutic strategy. be used in vivo, have a high specificity and can. The findings from these studies will allow us to identify the molecular and cellular mechanisms in the injured peritoneum which are addressed by our novel therapeutic strategy and to develop a targeted therapeutic approach using specific nanoparticles 3. To improve the pharmacokinetic and pharmacodynamic profile of of our antisense approach we will use nanoparticles. This approach will allow us to develop sophisticated targeting strategies and reduces the concentration of our antisense compound used.

在我们之前的实验中，我们能够在实验支持下证明PKCα是唯一在mpmc中表达的经典PKC亚型。PKCα蛋白的表达和激活在体外pd模拟条件下和体内长时间暴露于PDF后上调。而在WT小鼠中，慢性PDF会导致PM严重的形态、结构和功能改变，而Go6976对PKCα活性的功能阻断完全保护PM的结构和功能。PKCα缺乏显示出类似的益处，表明PKCα在PM紊乱中起关键作用。基于上一个资助期的这些发现，我们提出了一种新的治疗方法来预防pd -液诱导的炎症、新生血管生成和纤维化。1. 在透析期间，我们希望使用一种针对PKC异构体α的反义策略来抑制葡萄糖诱导的炎症、新血管生成和纤维化。特殊的反义方法防止了其他PKC抑制剂如Gö的副作用，这些抑制剂也可能影响多种蛋白激酶家族的多种激酶，并且通常具有其他药理作用，导致体外和体内不同的作用，独立于PKC抑制。2. 我们将分析我们的策略对腹膜不同细胞类型的治疗效果。我们从先前的研究中得到证据表明，间皮细胞是主要的治疗靶点。然而，我们之前也看到PKC-a在内皮细胞、巨噬细胞和成纤维细胞中发挥重要作用。因此，我们计划使用单核基因表达分析来确定我们的治疗策略。在体内使用，具有很高的特异性，可以。这些研究的发现将使我们能够确定受损腹膜的分子和细胞机制，这是我们的新治疗策略所解决的问题，并开发出一种使用特定纳米颗粒的靶向治疗方法。为了改善我们的反义方法的药代动力学和药效学特征，我们将使用纳米颗粒。这种方法将使我们能够开发复杂的靶向策略，并减少我们使用的反义化合物的浓度。