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.

这项研究旨在确定前列腺素信号传导在吻合泄漏中的作用，并测试支持愈合途径的介入策略。在腹部手术期间，外科医生对内脏器官的术中处理不可避免地会施加手术创伤，并且由此产生的组织损伤会影响常规器官功能。我们的小组拥有丰富的经验，研究了几种手术创伤引起的疾病的病理机制，包括术后肠胃，腹腔内粘附形成和吻合式泄漏。在该项目中，我们将探索吻合泄漏的分子机制，从而严重干扰结肠手术患者的康复。结直肠吻合泄漏（CAL）是一种可怕的手术并发症，对应于结直肠手术后所有死亡的三分之一。 CAL被认为是一个复杂的过程，涉及各种细胞反应。前列腺素（PG）是通过环氧酶（COX）从花生四烯酸产生的，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活性会大大增加CAL，因此在临床检查中经常会规避。为了评估驱动CAL的分子途径，我们通过转录分析分析了CAL，并将PGE2受体EP2和EP4鉴定为CAL的潜在介质。我们假设EP2或EP4信号传导调节吻合术的愈合，这些途径的干扰可能会介导Cal。由于EP2和EP4通过各种细胞类型表达并影响众多细胞过程，因此必须进行详细研究以识别CAL的分子机制以允许对其病理进行询问。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）和空间转录组学的CAL中PGE2和细胞因子产生的位置的鉴定。 ii）研究抑制固有的PGE2降解途径，EP2和EP4受体激动剂以及在两个不同CAL模型中抑制细胞特异性EP2和EP4信号传导，以识别预防CAL预防的潜在新途径。 iii）基于UPLC-MS的Oxylipin面板测定法对人吻合治疗剂和泄漏的血液样本进行了翻译分析，以确定COX衍生的介体模式，并潜在地与接受结直肠吻合手术的人类的愈合结果不同。总体而言，该项目所产生的数据集将提供对愈合和CAL鼠吻合的信号传导途径的新洞察力，重点是PGE2信号传导。本文生成的数据包括在单细胞水平上进行的全面转录表征，脂质分析，空间转录组学和CAL中的介入策略。 Together, these data will help to develop targeted drugs to prevent CAL prevention in the future.