Lonati Caterina, Schlegel Andrea, Battistin Michele, Merighi Riccardo, Carbonaro Margherita, Dongiovanni Paola, Leonardi Patrizia, Zanella Alberto, Dondossola Daniele
Center for Preclinical Research, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, 20122 Milan, Italy.
Hepatobiliary Unit, Careggi University Hospital, University of Florence, 50139 Florence, Italy.
Biomedicines. 2021 Oct 11;9(10):1444. doi: 10.3390/biomedicines9101444.
Hypothermic-oxygenated-machine-perfusion (HOPE) allows assessment/reconditioning of livers procured from high-risk donors before transplantation. Graft referral to HOPE mostly depends on surgeons' subjective judgment, as objective criteria are still insufficient. We investigated whether analysis of effluent fluids collected upon organ flush during static-cold-storage can improve selection criteria for HOPE utilization. Effluents were analyzed to determine cytolysis enzymes, metabolites, inflammation-related mediators, and damage-associated-molecular-patterns. Molecular profiles were assessed by unsupervised cluster analysis. Differences between "machine perfusion (MP)-yes" vs. "MP-no"; "brain-death (DBD) vs. donation-after-circulatory-death (DCD)"; "early-allograft-dysfunction (EAD)-yes" vs. "EAD-no" groups, as well as correlation between effluent variables and transplantation outcome, were investigated. Livers assigned to HOPE ( = 18) showed a different molecular profile relative to grafts transplanted without this procedure ( = 21, = 0.021). Increases in the inflammatory mediators PTX3 ( = 0.048), CXCL8/IL-8 ( = 0.017), TNF-α ( = 0.038), and ANGPTL4 ( = 0.010) were observed, whereas the anti-inflammatory cytokine IL-10 was reduced ( = 0.007). Peculiar inflammation, cell death, and coagulation signatures were observed in fluids collected from DCD livers compared to those from DBD grafts. AST ( = 0.034), ALT ( = 0.047), and LDH ( = 0.047) were higher in the "EAD-yes" compared to the "EAD-no" group. Cytolysis markers and hyaluronan correlated with recipient creatinine, AST, and ICU stay. The study demonstrates that effluent molecular analysis can provide directions about the use of HOPE.
低温氧合机器灌注（HOPE）可在移植前对来自高风险供体的肝脏进行评估/修复。肝脏是否提交给HOPE主要取决于外科医生的主观判断，因为客观标准仍不充分。我们研究了在静态冷藏期间器官冲洗时收集的流出液分析是否可以改进HOPE使用的选择标准。对流出液进行分析以确定细胞溶解酶、代谢物、炎症相关介质和损伤相关分子模式。通过无监督聚类分析评估分子谱。研究了“机器灌注（MP）-是”与“MP-否”；“脑死亡（DBD）与循环死亡后捐赠（DCD）”；“早期移植肝功能障碍（EAD）-是”与“EAD-否”组之间的差异，以及流出液变量与移植结果之间的相关性。分配给HOPE的肝脏（n = 18）与未进行此程序而移植的移植物（n = 21，P = 0.021）相比显示出不同的分子谱。观察到炎症介质PTX3（P = 0.048）、CXCL8/IL-8（P = 0.017）、TNF-α（P = 0.038）和ANGPTL4（P = 0.010）增加，而抗炎细胞因子IL-10减少（P = 0.007）。与来自DBD移植物的流出液相比，在从DCD肝脏收集的流出液中观察到特殊的炎症、细胞死亡和凝血特征。“EAD-是”组的AST（P = 0.034）、ALT（P = 0.047）和LDH（P = 0.047）高于“EAD-否”组。细胞溶解标志物和透明质酸与受者肌酐、AST和ICU住院时间相关。该研究表明，流出液分子分析可为HOPE的使用提供指导。
