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

在我们以前的实验中，我们在实验中证明了α是唯一在MPMC中表达的经典的PKC亚型。在体外模拟帕金森病条件下和体内长期接触PDF后，PKCα蛋白的表达和激活均上调。而在WT小鼠，慢性给予PDF可导致PM的形态、结构和功能发生严重变化，功能阻断PKCα活性，GO6976完全保护PM的结构和功能。蛋白激酶Cα缺乏也显示出类似的益处，提示蛋白激酶Cα在PM干扰中起着关键作用。基于这些发现，在上一个资金阶段，我们提出了一种新的治疗方法，以防止PD液诱导的炎症、新生血管生成和纤维化。1.在腹膜透析液中定期应用针对PKCα亚型的反义策略，以抑制葡萄糖诱导的炎症、新生血管生成和纤维化。这种特异的反义方法可以防止其他PKC抑制剂的副作用，例如Gö，Gö也可能影响来自各种蛋白激酶家族的多种激酶，并且通常具有其他药理作用，导致在体外和体内不依赖于PKC抑制而产生不同的效果。2.我们将分析我们的策略对不同类型腹膜细胞的治疗效果。我们从以前的研究中得到的证据表明，间皮细胞是主要的治疗目标。然而，我们也已经看到，PKC-a在内皮细胞、巨噬细胞和成纤维细胞中发挥着重要的作用。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，具有较高的特异性和可操作性。这些研究的结果将使我们能够确定我们新的治疗策略解决的受损腹膜中的分子和细胞机制，并开发使用特定纳米颗粒的靶向治疗方法3。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。

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.