Molecular mechanisms mediating colonic anastomotic leakage.

介导结肠吻合口漏的分子机制。

• 批准号: 534798626
• 负责人: Professor Dr. Sven Wehner
• 金额: --
• 依托单位: Klinik und Poliklinik für Allgemein-, Viszeral-, Thorax- und Gefäßchirurgie
• 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/534798626?language=en
• 关键词: Molecular; mechanisms; mediating; colonic; anastomotic

This study aims to identify the role of prostaglandin signalling in anastomotic leakage and to test interventional strategies to support healing pathways. During abdominal surgery, the surgeon's intraoperative handling of visceral organs unavoidably exerts a surgical trauma, and the resulting tissue damage can affect regular organ function. Our group has extensive experience studying the pathological mechanisms of several surgical trauma-induced disorders, including postoperative ileus, intra-abdominal adhesion formation and anastomotic leakage. In this project, we will explore the molecular mechanisms of anastomotic leakage that significantly disturb colonic surgery patients' recovery. Colorectal anastomotic leakage (CAL) is a dreaded surgical complication and corresponds to one-third of all deaths after colorectal surgery. CAL has been recognised as a complex process involving diverse cellular responses. Prostaglandins (PG) are produced from arachidonic acid via cyclooxygenases (COX) and are crucial for regulating wound healing processes. Consistently, inhibition of COX activity in a clinical setting is known to augment CAL considerably and is therefore often circumvented in the clinical workup. To assess the molecular pathways driving CAL, we analysed CAL in mice through transcriptional profiling and identified PGE2 receptors EP2 and EP4 as potential mediators of CAL. We hypothezised that EP2 or EP4 signalling regulates anastomotic healing and disturbances of these pathways are likely to mediate CAL. Since EP2 and EP4 are expressed by various cell types and influence numerous cellular processes, a detailed investigation is necessary to identify the molecular mechanisms of CAL to allow interrogation of its pathology. In this study, we have three main objectives: i) Identification of the cellular source and location of PGE2 and cytokine production in CAL by single-cell RNAseq, desorption electrospray mass spectrometry imaging (DESI) and spatial transcriptomics. ii) To study the inhibition of intrinsic PGE2 degrading pathways, EP2 and EP4 receptor agonists and inhibition of cell-specific EP2 and EP4 signalling in two different CAL models to identify potentially novel pathways in CAL prevention. iii) Translational analyses of human anastomotic healers and leakers blood samples by UPLC-MS-based oxylipin panel assay to determine COX-derived mediator patterns and potential association with different healing outcomes in humans undergoing colorectal anastomotic surgery. Overall, the datasets yielded by this project will provide novel insight into the signalling pathways of murine anastomosis during healing and CAL with a focus on PGE2 signalling. The data generated herein include comprehensive transcriptional characterisation at the single cell level, lipidomic profiling, spatial transcriptomics and interventional strategies in CAL. Together, these data will help to develop targeted drugs to prevent CAL prevention in the future.

本研究旨在确定前列腺素信号传导在吻合口瘘中的作用，并测试支持愈合途径的介入策略。在腹部手术中，外科医生术中处理内脏器官不可避免地会造成手术创伤，由此产生的组织损伤会影响器官的正常功能。我们小组在研究多种手术创伤引起的疾病的病理机制方面拥有丰富的经验，包括术后肠梗阻、腹腔内粘连形成和吻合口瘘。在这个项目中，我们将探讨严重影响结肠手术患者康复的吻合口瘘的分子机制。结直肠吻合口漏 (CAL) 是一种可怕的手术并发症，占结直肠手术后死亡人数的三分之一。 CAL 被认为是一个涉及多种细胞反应的复杂过程。前列腺素 (PG) 由花生四烯酸通过环加氧酶 (COX) 产生，对于调节伤口愈合过程至关重要。一致地，已知在临床环境中抑制 COX 活性可显着增强 CAL，因此在临床检查中经常被规避。为了评估驱动 CAL 的分子途径，我们通过转录谱分析了小鼠的 CAL，并确定 PGE2 受体 EP2 和 EP4 作为 CAL 的潜在介质。我们假设 EP2 或 EP4 信号传导调节吻合口愈合，并且这些通路的干扰可能介导 CAL。由于 EP2 和 EP4 由各种细胞类型表达并影响许多细胞过程，因此有必要进行详细的研究来确定 CAL 的分子机制，以便研究其病理学。在这项研究中，我们有三个主要目标：i) 通过单细胞 RNAseq、解吸电喷雾质谱成像 (DESI) 和空间转录组学鉴定 CAL 中 PGE2 和细胞因子产生的细胞来源和位置。 ii) 研究两种不同 CAL 模型中内在 PGE2 降解途径、EP2 和 EP4 受体激动剂的抑制作用以及细胞特异性 EP2 和 EP4 信号传导的抑制作用，以确定预防 CAL 的潜在新途径。 iii) 通过基于 UPLC-MS 的氧脂素小组测定对人类吻合治疗者和泄漏者血液样本进行转化分析，以确定 COX 衍生的介质模式以及与接受结直肠吻合手术的人类不同愈合结果的潜在关联。总体而言，该项目产生的数据集将为小鼠吻合在愈合和 CAL 过程中的信号通路提供新的见解，重点关注 PGE2 信号传导。本文生成的数据包括单细胞水平的综合转录表征、脂质组学分析、空间转录组学和 CAL 中的介入策略。总之，这些数据将有助于开发有针对性的药物来预防未来的 CAL。