Sorbini Monica, Carradori Tullia, Patrono Damiano, Togliatto Gabriele, Caorsi Cristiana, Vaisitti Tiziana, Mansouri Morteza, Delsedime Luisa, Vissio Elena, De Stefano Nicola, Papotti Mauro, Amoroso Antonio, Romagnoli Renato, Deaglio Silvia
Department of Medical Sciences, University of Turin, Turin, Italy.
General Surgery 2U-Liver Transplant Center, Department of Surgical Sciences, AOU Città della Salute e della Scienza di Torino, Turin, Italy.
Artif Organs. 2025 Apr;49(4):649-662. doi: 10.1111/aor.14910. Epub 2024 Nov 18.
Liver transplantation (LT) is still limited by organ shortage and post-transplant monitoring issues. While machine perfusion techniques allow for improving organ preservation, biomarkers like donor-derived cell-free DNA (dd-cfDNA) and mitochondrial cfDNA (mt-cfDNA) may provide insights into graft injury and viability pre- and post-LT.
A prospective observational cohort study was conducted on LT recipients (n = 45) to evaluate dd-cfDNA as a biomarker of graft dysfunction during the first 6 months after LT. Dd-cfDNA was quantified on blood samples collected pre-LT and post-LT using droplet digital PCR. In livers undergoing dual hypothermic oxygenated machine perfusion (D-HOPE), total cfDNA and mt-cfDNA levels were measured on perfusate samples collected at 30-min intervals. Correlations with graft function and clinical outcomes were assessed.
Dd-cfDNA levels peaked post-LT and correlated with transaminase levels and histological injury severity. The longitudinal assessment showed that postoperative complications and rejection were associated with an increase in dd-cfDNA levels. Mt-cfDNA levels in D-HOPE perfusate correlated with graft function parameters post-LT and were higher in patients with early allograft dysfunction and severe complications.
This study confirms dd-cfDNA as a marker of graft injury after LT and suggests that perfusate mt-cfDNA levels during D-HOPE correlate with graft function and post-transplant clinical outcome. Integration of these tests into clinical practice may improve transplant management and viability assessment during hypothermic perfusion.
肝移植（LT）仍受器官短缺和移植后监测问题的限制。虽然机器灌注技术有助于改善器官保存，但诸如供体来源的游离DNA（dd-cfDNA）和线粒体cfDNA（mt-cfDNA）等生物标志物可能为肝移植前后的移植物损伤和活力提供见解。
对LT受者（n = 45）进行了一项前瞻性观察队列研究，以评估dd-cfDNA作为LT后前6个月移植物功能障碍的生物标志物。使用液滴数字PCR对LT前和LT后采集的血样中的dd-cfDNA进行定量。在接受双低温氧合机器灌注（D-HOPE）的肝脏中，每隔30分钟采集一次灌注液样本，测量总cfDNA和mt-cfDNA水平。评估与移植物功能和临床结果的相关性。
dd-cfDNA水平在LT后达到峰值，并与转氨酶水平和组织学损伤严重程度相关。纵向评估表明，术后并发症和排斥反应与dd-cfDNA水平升高有关。D-HOPE灌注液中的mt-cfDNA水平与LT后的移植物功能参数相关，在早期移植物功能障碍和严重并发症患者中更高。
本研究证实dd-cfDNA是LT后移植物损伤的标志物，并表明D-HOPE期间灌注液mt-cfDNA水平与移植物功能和移植后临床结果相关。将这些检测纳入临床实践可能会改善低温灌注期间的移植管理和活力评估。
