Preclinical efficacy and safety evaluation of human embryonic stem cell-derived functional hepatocytes for a clinical trial in acute liver failure

Abstract Background Acute liver failure (ALF) is a life-threatening syndrome characterized by rapid deterioration of liver function, resulting in high mortality and posing a substantial global health burden. Human embryonic stem cells (hESCs) possess unlimited self-renewal capacity and pluripotent differentiation potential. Transplantation of hESC-derived hepatocyte-like cells (HPLCs) represents a promising therapeutic strategy for ALF. Methods A good manufacturing practice (GMP)-compliant differentiation process was developed to generate HPLCs from hESCs, and their biological characteristics and functional properties were systematically evaluated. A comprehensive series of preclinical safety and efficacy studies was performed, including dose-escalation experiments, biodistribution analysis, comparative evaluation of administration routes, and carcinogenicity testing. The therapeutic efficacy and safety of HPLCs were assessed in a fatal rat model of D-galactosamine (D-gal) and lipopolysaccharide (LPS)-induced ALF. In addition, the HPLCs underwent quality evaluation by the National Institutes for Food and Drug Control (NIFDC), and an independent safety assessment was conducted. Results A high-efficiency system was established for the generation of qualified, clinical-grade HPLCs from hESCs under GMP-compliant conditions. The HPLCs exhibited multiple mature hepatocyte functions, including carbohydrate and lipid metabolism, hepatic synthetic and storage functions, inducible cytochrome P450 activity, albumin secretion, and urea production. The HPLCs met the certification standards of the NIFDC of China. Transplantation of HPLCs significantly improved survival in ALF rats, with survival rates of 72.4% following tail vein injection and 66.7% following intraperitoneal injection, compared with 6.67% in the control group. HPLC transplantation also promoted recovery of liver function, as reflected by improvements in biochemical and coagulation parameters. Preclinical safety evaluations confirmed the biosafety of HPLCs, with no evidence of acute toxicity or tumorigenicity. A Phase I clinical trial (ChiCTR2100052988) for the treatment of ALF and acute-on-chronic liver failure (ACLF) has been approved by the National Health Commission of the People’s Republic of China (Filing Number: MR-43-21-014643) and has been initiated. Conclusions A novel multistage, GMP-compliant process was developed for the effective and reproducible differentiation of hESCs into hepatocytes. The resulting HPLCs demonstrated robust hepatocyte functions, therapeutic efficacy in an ALF animal model, and favorable biosafety profiles. These findings support the clinical translation of HPLCs, with an ongoing Phase I clinical trial designed to evaluate their safety and feasibility in patients with ALF and ACLF.