DataSheet_1_Patient-Derived Avatar Mouse Model to Predict the Liver Immune Homeostasis of Long-Term Stable Liver Transplant Patients.pdf

Although rejection or tolerance can occur in liver transplantation (LT) patients, there are no reliable non-invasive methods for predicting immune homeostasis. In this study, we developed a humanized mouse model to predict liver immune homeostasis in patients who underwent LT. The patient-derived avatar model was developed by injecting peripheral blood mononuclear cells from healthy controls (HCs) or LT patients with stable, rejection, or tolerance into NOD.Cg-PrkdcscidIL2rgtm1Wjl/SzJ (NSG) mice, followed by injection of human hepatic stellate cells and Carbone tetrachloride (CCl4). After 7 weeks, the patient’s T-cell engraftment and liver inflammation in the avatar model were evaluated and compared with the liver histology of LT patients. Changes in liver inflammation following treatment with tacrolimus and/or biguanide derivatives were also examined. The C-X-C Motif Chemokine Receptor 3 (CXCR3)-dependently engrafted patient T cells led to differences in liver inflammation in our model according to the status of LT patients. The livers of avatar models from rejection patients had severe inflammation with more T helper 17 cells and fewer regulatory T cells compared to those of models from tolerance and HCs showing only mild inflammation. Moreover, our model classified stable post-LT patients into severe and mild inflammation groups, which correlated well with liver immunity in these patients. Our models revealed alleviation of inflammation after combination treatment with tacrolimus and biguanide derivatives or monotherapy. Consequently, using our new patient-derived avatar model, we predicted liver immune homeostasis in patients with stable LT without biopsy. Moreover, our avatar model may be useful for preclinical analysis to evaluate treatment responses while reducing risks to patients.

尽管肝移植（Liver Transplantation, LT）患者可能出现排斥反应或免疫耐受，但目前尚无可靠的非侵入性方法用于预测其免疫稳态。本研究构建了一种人源化小鼠模型，用于预测接受肝移植患者的肝脏免疫稳态。该患者源性替身模型的构建流程为：将健康对照（Healthy Controls, HCs）或处于稳定状态、发生排斥反应或免疫耐受的肝移植患者的外周血单个核细胞（peripheral blood mononuclear cells, PBMC）注射至NOD.Cg-PrkdcscidIL2rgtm1Wjl/SzJ（NSG）小鼠体内，随后注射人肝星状细胞与四氯化碳（Carbon tetrachloride, CCl4，原文存在Carbone笔误）；造模7周后，评估该替身模型中的患者T细胞植入率与肝脏炎症程度，并与肝移植患者的肝脏组织病理学结果进行对比，同时检测他克莫司（Tacrolimus）和/或双胍类衍生物治疗后肝脏炎症的变化情况。基于肝移植患者的不同临床状态，依赖C-X-C基序趋化因子受体3（CXCR3）植入的患者T细胞可导致本模型中肝脏炎症程度产生差异：与来自免疫耐受患者及健康对照的替身模型（仅表现为轻度肝脏炎症）相比，来自排斥反应患者的替身模型肝脏存在严重炎症，且其体内辅助性T细胞17（T helper 17, Th17）数量更多、调节性T细胞（regulatory T cell, Treg）数量更少。此外，本模型可将术后病情稳定的肝移植患者分为重度与轻度炎症两组，且该分类结果与这些患者的肝脏免疫状态显著相关。模型结果显示，采用他克莫司与双胍类衍生物联合治疗或单一疗法均可减轻肝脏炎症。因此，借助本研究构建的新型患者源性替身模型，我们无需通过活检即可预测病情稳定的肝移植患者的肝脏免疫稳态；此外，该替身模型可用于临床前分析以评估治疗反应，同时降低患者面临的风险。