Investigation of Clinical Safety of Human iPS Cell-Derived Liver Organoid Transplantation to Infantile Patients in Porcine Model

Abstract

Transplantation of liver organoids has been investigated as a treatment alternative to liver transplantation for chronic liver disease. Transportal approach can be considered as a method of delivering organoids to the liver. It is important to set the allowable organoid amount and verify translocation by intraportal transplantation. We first examined the transplantation tolerance and translocation of porcine fetal liver-derived allogeneic organoids using piglets. Fetal liver-derived organoids generated from the Kusabira Orange-transduced pig were transplanted to the 10-day-old piglet liver through the left branch of the portal vein. All recipients survived without any observable adverse events. In contrast, both local and main portal pressures increased transiently during transplantation. In necropsy samples, Kusabira Orange-positive donor cells were detected primarily in the target lobe of the liver and partly in other areas, including the lungs and brain. As we confirmed the transplantation allowance by porcine fetal liver-derived organoids, we performed intraportal transplantation of human-induced pluripotent stem cell (iPSC)-derived liver organoid, which we plan to use in clinical trials, and portal pressure and translocation were investigated. Human iPSC-derived liver organoids were transplanted into the same 10-day-old piglet. Portal hypertension and translocation of human iPSC-derived liver organoids to the lungs were observed in one of two transplanted animals. Translocation occurred in the piglet in which patent ductus venosus (PDV) was observed. Therefore, a 28-day-old piglet capable of surgically ligating PDV was used, and after the PDV was ligated, human iPSC-derived liver organoids with the amount of which is scheduled in clinical trials were transplanted. This procedure inhibited the translocation of human iPSC-derived liver organoids to extrahepatic sites without no portal hypertension. In conclusion, human iPSC-derived liver organoids can be safely transplanted through the portal vein. Ligation of the ductus venosus prior to transplantation was effective in inhibiting extrahepatic translocation in newborns and infants.

Characterization and Treatment Responsiveness of Genetically Engineered Ornithine Transcarbamylase-Deficient Pig

“…However, although liver transplantation is an effective treatment, patients have to take immunosuppressive drugs on a lifelong basis, and risk of rejection and need for dietary restrictions exist long after transplantation. While cell transplantation and gene therapy are being examined as novel treatments [ 5 , 8 , 9 , 10 ], their safety and therapeutic efficacies have not yet been determined. From this point of view, human disease models in which clinical equivalent treatments are executable are preferable for the development of novel medical technologies.…”

“…Thus, the severe fulminant OTCD model was established. The well-known OTCD model animals, sparse-fur (spf/1) mice (ID156, Balb/c background spontaneous mutation mice) show 1/6 the level of OTC activity (wild-type mice: 317.8 ± 51.5 nmol/min/mg, OTCD mice: 53.7 ± 33.5 nmol/min/mg, [mean ± SD, n = 3], unpublished data) and survive for approximately two months (median, 47 days; range: 20–52) [ 5 , 10 ]. In our previous study, hepatocyte transplantation two days after birth significantly prolonged the survival time in OTCD mice [ 10 ].…”

“…Anesthesia and analgesia for hysterotomy and catheter insertion were performed according to standard veterinary medicine protocols [ 5 ]. Briefly, pigs were sedated by intramuscular injection of 0.5 mg/kg mafoprazine mesylate (Mafropane ® ; DS Pharma Animal Health Co., Ltd., Osaka, Japan) followed by an intravenous injection of 1.5 mg/kg sodium thiopental (Ravonal ® ; NIPRO ES Pharma).…”

Emerging liver organoid platforms and technologies

“…To note, the system described by Taniguchi’s group has achieved remarkable scalability (Takebe et al 2017 ) and also demonstrated the possibility to use different cell types derived from a single iPSC clone (Zhang et al 2018b ). This system has now been validated in larger porcine models for clinical safety with portal vein transplantation procedures and shows great promise for human clinical trials in the near future (Tsuchida et al 2020 ).…”

Hepatitis B virus infection modeling using multi-cellular organoids derived from human induced pluripotent stem cells

“…Moreover, the potential risks associated with genetic modification, tumorigenesis, and transplantation techniques should be emphasized in light of individual and societal values. In the context of continuing investigation regarding clinical concerns[ 79 , 80 ], quality-controlled and personalized LOs from authorized patient-derived cell banks are expected to be used routinely in the future.…”