Xue Wei-Song, Zhang Hao-Jie, Ke Jing-Jing, Fu Yu, Peng Qing, Li Li, Gao Yi, Zhong Ke-Bo
General Surgery Center, Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Guangzhou Clinical Research and Transformation Center for Artificial Liver, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, China; Department of General Surgery & Guangdong Provincial Key Laboratory of Precision Medicine for Gastrointestinal Tumor, Nanfang Hospital, The First School of Clinical Medicine, Southern Medical University, Guangzhou 510515, China.
General Surgery Center, Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Guangzhou Clinical Research and Transformation Center for Artificial Liver, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, China.
Hepatobiliary Pancreat Dis Int. 2023 Jun;22(3):270-281. doi: 10.1016/j.hbpd.2022.06.012. Epub 2022 Jun 30.
Preventing heterologous protein influx in patients is important when using xenogeneic bioartificial livers (BALs) to treat liver failure. The development of transgenic porcine livers synthesizing human proteins is a promising approach in this regard. Here, we evaluated the safety and efficacy of a transgenic porcine liver synthesizing human albumin (hALB) and coagulation factor VII (hFVII) within a bioartificial system.
Tibetan miniature pigs were randomly subjected to different interventions after surgery-induced partially ischemic liver failure. Group A (n = 4) was subjected to basic treatment; group B (n = 4) was to standard medical treatment and wild-type porcine BAL perfusion, and group C (n = 2) was to standard medical treatment and transgenic BAL perfusion. Biochemical parameters, coagulation status, survival time, and pathological changes were determined. Expressions of hALB and hFVII were detected using immunohistochemistry and enzyme-linked immunosorbent assays.
The survival time in group A was 9.75 ± 1.26 days; this was shorter than that in both perfused groups, in which all animals reached an endpoint of 12 days (P = 0.006). Ammonia, bilirubin, and lactate levels were significantly decreased, whereas albumin and fibrinogen levels were increased after perfusion (all P < 0.05). hALB and hFVII were detected in transgenic BAL-perfused pig serum and ex vivo in the liver tissues.
The humanized transgenic pig livers could synthesize and secrete hALB and hFVII ex vivo in a whole organ-based bioartificial system, while maintaining their metabolism, detoxification, transformation, and excretion functions, which were comparable to those observed in wild-type porcine livers. Therefore, the use of transgenic bioartificial whole livers is expected to become a new approach in treating acute liver failure.
在使用异种生物人工肝（BAL）治疗肝衰竭时，防止患者体内异源蛋白流入非常重要。在这方面，开发合成人类蛋白质的转基因猪肝是一种很有前景的方法。在此，我们在生物人工系统中评估了合成人白蛋白（hALB）和凝血因子VII（hFVII）的转基因猪肝的安全性和有效性。
手术诱导部分缺血性肝衰竭后，将藏猪随机进行不同干预。A组（n = 4）接受基础治疗；B组（n = 4）接受标准药物治疗并进行野生型猪BAL灌注，C组（n = 2）接受标准药物治疗并进行转基因BAL灌注。测定生化参数、凝血状态、生存时间和病理变化。采用免疫组织化学和酶联免疫吸附试验检测hALB和hFVII的表达。
A组的生存时间为9.75±1.26天；这比两个灌注组都短，两个灌注组所有动物均达到12天的终点（P = 0.006）。灌注后氨、胆红素和乳酸水平显著降低，而白蛋白和纤维蛋白原水平升高（所有P < 0.05）。在转基因BAL灌注猪血清和肝组织体外检测到hALB和hFVII。
人源化转基因猪肝可以在基于全器官的生物人工系统中体外合成和分泌hALB和hFVII，同时保持其代谢、解毒、转化和排泄功能，这些功能与野生型猪肝相当。因此，使用转基因生物人工全肝有望成为治疗急性肝衰竭的一种新方法。
