Importance of co-expression and co-activation of AT1, thromboxane A2 and EGF(HER1) receptors for angiotensin II-mediated signaling in vascular cells: analysis of molecular and functional interactions - part 2

AT1、血栓素 A2 和 EGF(HER1) 受体的共表达和共激活对于血管细胞中血管紧张素 II 介导的信号传导的重要性：分子和功能相互作用的分析 - 第 2 部分

• 批准号: 422875742
• 负责人: Professor Dr. Ralf Benndorf
• 金额: --
• 依托单位: Institut für Pharmazie
• 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/422875742?language=en
• 关键词: Importance; co; expression; activation; AT1

Angiotensin II (Ang II) induces endothelial dysfunction, vascular remodeling, and atherosclerosis as well as abdominal aortic aneurysm formation through activation of the AT1 receptor (AT1R). With regard to the pathological effects of Ang II in vascular cells, it is becoming increasingly clear that only the parallel activation of additional receptor types triggers full-scale Ang II-mediated signal transduction.In the first phase of the project, we demonstrated, for example, that EGF receptors (EGFR) or thromboxane A2 receptors (TP) expressed in vascular smooth muscle cells promote high-fat diet-induced vascular changes and atherosclerosis. In addition, our preliminary work reveals molecular and functional interactions of AT1R, TP, and EGFR with potential relevance to signal transduction, gene expression, and vascular cell homeostasis. Therefore, the proposed second phase of the project will further investigate how the interactions of the receptors AT1R, TP, and EGFR affect Ang II-induced cellular signal transduction, subsequent control of gene expression, and cell physiological consequences in vascular cells. To test this hypothesis, work packages (WP) will be addressed at the levels of (i) biophysical receptor interaction, (ii) cellular interaction of downstream signaling pathways, and (iii) cell physiological consequences of receptor interaction in vitro and in vivo. Continuing our cell biological approach and using validated FRET microscopy methods, mechanisms of receptor-receptor interaction will be investigated as well as whether AT1R-EGFR-TP triple complexes form in living cells. In addition, the Ang II-induced signalosome of vascular cells will be analyzed with particular emphasis on receptor-receptor interactions using mass spectrometric methods followed by validating Western Blot analyses (WP1). In addition, the characterization of receptor interaction with respect to intracellular signaling pathways of SRF transcriptional control will be continued (WP2), an analysis of receptor interaction for transcriptome regulation in human vascular cells will be pursued (WP3), and an experimental validation of bioinformatic predictions of (patho)physiological changes in human vascular cells will be performed (WP4 and WP5). Ultimately, the therapeutic potential of pharmacological blockade of EGFR and TP for Ang II-induced aortic atherogenesis and aneurysm formation, cardiac hypertrophy and mortality in ApoE knockout mice will be tested in vivo (AP6). We anticipate that the project will help clarify the importance of functional and molecular interactions between AT1R, TP, and EGFR for Ang II-mediated effects on vascular cells. By improving mechanistic understanding, it is expected to support the development of rational pharmacological therapeutic strategies for vascular diseases.

血管紧张素II(Ang II)通过激活AT1受体(AT1R)诱导内皮功能障碍、血管重构、动脉粥样硬化和腹主动脉瘤的形成。关于Ang II在血管细胞中的病理作用，越来越清楚的是，只有其他受体类型的平行激活才能触发全面的Ang II介导的信号转导。在项目的第一阶段，我们证明了血管平滑肌细胞表达的EGF受体(EGFR)或血栓素A2受体(TP)促进了高脂饮食诱导的血管变化和动脉粥样硬化。此外，我们的初步工作揭示了AT1R、TP和EGFR的分子和功能相互作用，可能与信号转导、基因表达和血管细胞动态平衡相关。因此，该项目的第二阶段将进一步研究AT1R、TP和EGFR受体之间的相互作用如何影响Ang II诱导的细胞信号转导，随后的基因表达调控，以及血管细胞中的细胞生理后果。为了验证这一假设，工作包(WP)将从(I)生物物理受体相互作用，(Ii)下游信号通路的细胞相互作用，以及(Iii)体外和体内受体相互作用的细胞生理后果的水平进行处理。继续我们的细胞生物学方法，并使用经过验证的FRET显微镜方法，将研究受体-受体相互作用的机制以及AT1R-EGFR-TP三重复合体是否在活细胞中形成。此外，将使用质谱学方法分析血管紧张素Ⅱ诱导的血管细胞信号体，特别强调受体-受体的相互作用，然后验证Western Blot分析(WP1)。此外，关于SRF转录调控的细胞内信号通路的受体相互作用的特征将继续(WP2)，将继续对人类血管细胞中转录组调节的受体相互作用进行分析(WP3)，并将对人类血管细胞(病理性)生理变化的生物信息学预测进行实验验证(WP4和WP5)。最终，将在体内测试EGFR和TP的药理阻断对Ang II诱导的ApoE基因敲除小鼠的动脉粥样硬化形成和动脉瘤形成、心肌肥厚和死亡率的治疗潜力(AP6)。我们预计该项目将有助于阐明AT1R、TP和EGFR之间的功能和分子相互作用在Ang II介导的血管细胞效应中的重要性。通过提高对机制的理解，有望为开发合理的血管疾病药物治疗策略提供支持。