Rimondi Erika, Melloni Elisabetta, Secchiero Paola, Braidotti Stefania, Maximova Natalia, Marcuzzi Annalisa
Department of Translational Medicine and LTTA Centre, University of Ferrara, Ferrara, Italy.
Department of Pediatrics, Pediatrics, Bone Marrow Transplant Unit, Institute for Maternal and Child Health-IRCCS Burlo Garofolo, Trieste, Italy.
J Exp Pharmacol. 2025 Sep 4;17:613-623. doi: 10.2147/JEP.S534739. eCollection 2025.
Acute graft-versus-host disease (aGVHD) is a significant cause of death in recipients of allogeneic hematopoietic stem cell transplantation. In this type of graft, the intestine is particularly affected, with the loss of intestinal barrier integrity playing a key role in its onset. In this scenario, the aim of the present research was to evaluate defibrotide, a heparin-like compound, marked for severe veno-occlusive disease, as an innovative therapeutic approach for restoring intestinal barrier integrity using an in vitro model and analyzing aGVHD patients' sera and clinical data.
Using an in vitro model of colon epithelium, we evaluated the modulation of tight junction proteins after defibrotide treatment, in basal condition or in presence of an inflammatory stimulus, by immunocytochemical and Western blotting analysis. Moreover, the study involved two patients with grade IV acute multisystem GVHD with great gastrointestinal compromission. Patients' sera were collected during the acute phase and remission of intestinal aGVHD and employed for the evaluation of a panel of 27 inflammatory cytokines using a Multiplex approach.
Defibrotide treatment significantly increased the protein expression of Zonulin-1 and Occludin (untreated vs treated with 200 μg/mL, p<0.01). In culture conditions mimicking inflammation, defibrotide countered the reduction of Occludin and Claudin-3 while preserving Zonulin-1 levels. Serum cytokine analysis of two patients receiving defibrotide for aGVHD showed significantly higher cytokine levels (IL-7, MIP-1β, IP-10, G-CSF, Eotaxin, IL-6) during the acute phase compared to remission after defibrotide treatment.
These findings suggest a potential therapeutic role for defibrotide in managing intestinal aGVHD by improving epithelial barrier integrity and reducing inflammation-related damage.
急性移植物抗宿主病（aGVHD）是异基因造血干细胞移植受者死亡的重要原因。在这类移植中，肠道尤其易受影响，肠道屏障完整性的丧失在其发病过程中起关键作用。在此背景下，本研究旨在评估去纤苷（一种用于严重肝静脉闭塞病的类肝素化合物），作为一种创新的治疗方法，通过体外模型以及分析aGVHD患者的血清和临床数据来恢复肠道屏障完整性。
使用结肠上皮体外模型，我们通过免疫细胞化学和蛋白质印迹分析，评估了在基础条件下或存在炎症刺激时去纤苷治疗后紧密连接蛋白的调节情况。此外，该研究纳入了两名患有IV级急性多系统GVHD且胃肠道严重受损的患者。在肠道aGVHD的急性期和缓解期收集患者血清，并采用多重检测方法评估一组27种炎性细胞因子。
去纤苷治疗显著增加了闭合蛋白-1和闭合蛋白的蛋白表达（未治疗组与用200μg/mL治疗组相比，p<0.01）。在模拟炎症的培养条件下，去纤苷对抗了闭合蛋白和紧密连接蛋白-3的减少，同时维持了闭合蛋白-1的水平。两名接受去纤苷治疗aGVHD的患者的血清细胞因子分析显示，急性期的细胞因子水平（IL-7、MIP-1β、IP-10、G-CSF、嗜酸性粒细胞趋化因子、IL-6）显著高于去纤苷治疗后的缓解期。
这些发现表明去纤苷在通过改善上皮屏障完整性和减少炎症相关损伤来治疗肠道aGVHD方面具有潜在的治疗作用。
