Comparison of competing ex vivo preservation techniques and investigation of the mechanisms of graft recovery during ex-vivo perfusion in marginal kidney grafts

边缘肾移植物离体灌注过程中竞争性离体保存技术的比较和移植物恢复机制的研究

基本信息

批准号：
425903082
负责人：
Dr. Laura Ioana Mazilescu
金额：
--
依托单位：
Klinik für Allgemeinchirurgie, Viszeral- und Transplantationschirurgie
依托单位国家：
德国
项目类别：
Research Fellowships
财政年份：
2019
资助国家：
德国
起止时间：
2018-12-31 至 2021-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/425903082?language=en
关键词：
Comparison competing ex vivo preservation

项目摘要

Kidney transplantation represents the treatment of choice for end-stage renal disease. The enormous gap between supply and demand has lead to the more frequent use of organs from extended criteria and donation after circulatory death. A major problem with these marginal grafts is delayed graft function due to an increased susceptibility to ischemia and reperfusion injury, which leads to decreased long-term function. Therefore, the transplant community has focused its efforts on investigating methods for improving the function of these grafts. Normothermic ex-vivo kidney perfusion and oxygenated hypothermic perfusion are promising options to achieve that. In a porcine model of donation after circulatory death, studies have shown that marginal grafts subjected to normothermic ex-vivo kidney perfusion instead of the traditional cold storage demonstrate improved function of the grafts. Also, there is evidence from other animal studies that oxygenated hypothermic perfusion is superior to cold storage. The objective of the present research project is to investigate which method offers the best option for kidney preservation and which mechanisms are involved in the improved donation after circulatory death graft function in perfused grafts over static cold storage. We hypothesize that providing oxygen to the grafts prevents anaerobic metabolism, which results in a reduction in mediators involved in ischemia-reperfusion. However, other novel pathways may also be implicated. Therefore, we will initiate an unbiased examination of gene expression through microarray analysis, to then validate the results using quantitative polymerase chain reaction and then correlate these results with appropriate protein expression studies such as through enzyme-linked immunosorbent assays and Western blots. Also, potential biomarkers that correlate with improved graft function can be identified and these markers may then be utilized to assess the function and health of marginal grafts undergoing ex-vivo perfusion prior to transplant in patients. Finally, we will investigate whether a combination of oxygenated hypothermic perfusion and normothermic ex-vivo kidney perfusion offers a better preservation method in marginal grafts. The results of this study will support the translation of oxygenated hypothermic perfusion and normothermic ex-vivo kidney perfusion to human studies, and a personalized approach could be adopted as marginal grafts deemed unfit for transplant may be subjected to oxygenated hypothermic perfusion and normothermic ex-vivo kidney perfusion for restoration, improvement and assessment of graft function prior to transplantation.

肾脏移植代表终末期肾脏疾病的选择治疗。供需之间的巨大差距导致循环死亡后延长标准和捐赠中更频繁地使用器官。这些边缘移植物的一个主要问题是由于缺血和再灌注损伤的敏感性增加而导致移植物功能延迟，从而导致长期功能降低。因此，移植社区的重点是研究改善这些移植物功能的方法。常规的前体肾脏灌注和氧化的低温灌注是实现这一目标的有希望的选择。在循环死亡后的猪捐赠模型中，研究表明，经受过正常热的前体肾脏灌注而不是传统冷藏的边缘移植物表明移植物的功能提高了。此外，还有其他动物研究的证据表明，氧化的低温灌注优于冷藏。本研究项目的目的是调查哪种方法为肾脏保存提供了最佳选择，以及在循环死亡移植物功能促进的移植物在灌注移植物中的循环死亡移植物功能而在静态冷存储中涉及哪些机制。我们假设向移植物提供氧气可防止厌氧代谢，从而导致参与缺血 - 重新灌注的介体的减少。但是，其他新颖的途径也可能涉及。因此，我们将通过微阵列分析启动对基因表达的无偏见，然后使用定量聚合酶链反应验证结果，然后将这些结果与适当的蛋白质表达研究（例如通过酶联免疫吸附测定法和蛋白质印刷）相关联。同样，可以鉴定与改善的移植功能相关的潜在生物标志物，然后可以利用这些标记来评估患者移植前接受过前体灌注的边缘移植物的功能和健康。最后，我们将研究含氧的低温灌注和常规的前体肾脏灌注是否提供了更好的保存方法。这项研究的结果将支持对人类研究的氧化体温过性灌注和正常热的肾脏灌注的翻译，并且可以采用个性化方法，因为认为不适合移植的边缘移植物可以接受氧气的含氧量的低渗灌注，并在恢复过程中进行氧化型灌注，并评估恢复术，并评估灌肠的转移。