Importance of the EGF receptor and the smooth muscle-endothelial interaction for the effects of obesity-associated milieu changes on vascular cells.

肥胖症的发生率不断增加，这在很大程度上导致了心血管疾病的流行。肥胖相关的环境变化（OAMC）可以直接影响血管壁的稳态。前期的工作表明表皮生长因子受体（EGFR）作为血管信息处理中的信号整合者的重要作用。我们的数据表明，血管肌肉（VSMC）-EGFR有助于高脂饮食（HFD）诱导的血管和肾脏变化。EGFR/ErbB 2-ROCK-MRTF-SRF信号轴的增强和线粒体功能障碍在此起重要作用。此外，VSMC和内皮细胞（EC）之间存在VSMC-EGFR依赖性相互作用。另一方面，EC-EGFR对血管功能本身以及HFD诱导的损伤的重要性较小且部分相反。在原代VSMC中观察到的EGFR介导的TGFβ和CTGF表达代表了可能的相互作用机制。在初步的实验中，我们能够表明，EGF对原代VSMC（野生型，但不是EGFR-KO）的转录组的影响是由葡萄糖增强，并有，除其他事项外，骨桥蛋白的表达方面的协同作用，也是一个候选人的VSMC-EC interaction.Objective：分析的重要性，表皮生长因子受体的OAMC对原代VSMC和EC和细胞相互作用的影响。OAMC的成分包括代谢应激（葡萄糖、游离脂肪酸）和体液应激（血管紧张素II、去甲肾上腺素）。在这里，VSMC和EC之间发生VSMC-EGFR依赖性相互作用。EC的改变还可以包括EC-EGFR的积极作用的减弱和受影响的EC对VSMC的有害反馈。的相互作用假说，EGFR的参与和它的相关性，为人类血管cells现在必须检查和信息transmittingidentified.Action概念的机制：我们将进行比较实验与鼠和人的主要VSMC和EC在单和共培养。小鼠模型允许通过比较来自共培养的EGFR-KO动物和WT动物的细胞以细胞特异性方式研究EGFR的作用。转基因小鼠模型的细胞对病理生理过程的机制理解做出了重大贡献。然而，转移到人细胞通常是不可能的，因此必须使用离体原代人细胞培养系统。然后在人模型中使用EGFR抑制剂。计划进行以下工作包以检验相应的假设：1.增殖、衰老、细胞死亡; 2.能量稳态; 3.细胞外基质; 4.炎症; 5和6.细胞间和细胞内信号转导。在信息传递方面，通过媒体分析和干预实验研究了细胞间的信息传递。

The increasing frequency of obesity contributes significantly to the prevalence of cardiovascular diseases. Obesity-associated milieu changes (OAMC) can directly affect vascular wall homeostasis. Work from the previous project period shows the important role of EGF receptor (EGFR), as a signal integrator in vascular information processing. Our data show that the vascular muscle (VSMC)-EGFR contributes to high-fat diet (HFD)-induced vascular and renal changes. The potentiation of the EGFR/ErbB2-ROCK-MRTF-SRF signaling axis and mitochondrial dysfunction play an important role here. In addition, there is a VSMC-EGFR-dependent interaction between VSMC and endothelial cells (EC). The EC-EGFR, on the other hand, is of lesser and partly contrary importance for vascular function per se, as well as for HFD-induced damage. The observed EGFR-mediated expression of TGFβ and CTGF in primary VSMC represents a possible mechanism of interaction. In preliminary experiments we were able to show that the effect of EGF on the transcriptome of primary VSMC (wildtype but not EGFR-KO) is enhanced by glucose and that there is, among other things, a synergism with regard to the expression of osteopontin, also a candidate for the VSMC-EC interaction.Objective: Analyzing the importance of EGFR for the effects of OAMC on primary VSMC and EC and of cellular interaction. The components of OAMC to be examined include metabolic stress (glucose, free fatty acids) and humoral stress (angiotensin II, norepinephrine).Action need: Our main working hypotheses state that the VSMC-EGFR is necessary for pathological vascular changes in VSMC and EC. Here, a VSMC-EGFR-dependent interaction between VSMC and EC takes place. Resulting alterations of EC can also include a weakening of positive effects of EC-EGFR and detrimental feedback of affected EC on VSMC. The interaction hypothesis, the involvement of EGFR and its relevance for human vascular cells must now be examined and the mechanisms of information transfer identified.Action concept: We will carry out comparative experiments with murine and human primary VSMC and EC in mono- and co-culture. The mouse models allow the role of EGFR to be investigated in a cell-specific manner by comparing cells from EGFR-KO animals and WT animals in coculture. Cells of transgenic mouse models have contributed significantly to the mechanistic understanding of pathophysiological processes. However, the transfer to human cells was often not possible, so that the use of ex vivo primary human cell culture systems is necessary. EGFR inhibitors are then used in the human model. The following work packages for testing the corresponding hypotheses are planned: 1. proliferation, senescence, cell death; 2. energy homeostasis; 3. extracellular matrix; 4. inflammation; 5&6. inter- and intracellular signal transduction. With regard to information transfer, intercellular communication is exam-ined by means of media analysis and intervention experiments.