(T1085) SINGLE-CELL TRANSCRIPTOMIC ROADMAP TO HUMAN INDUCED HEPATOCYTE-LIKE CELLS REVEALS A LIVER DEVELOPMENTAL TRAJECTORY DURING LINEAGE REPROGRAMMING
Abstract: Hepatocytes are crucial for drug screening, disease modeling, and clinical trans-plantation, yet generating functional hepatocytes in vitro is challenging due to the difficulty of establishing their authentic gene regulatory networks (GRNs). We have previously developed a two-step lineage reprogramming strategy to generate func-tionally competent human induced hepatocytes (hiHeps), providing an effective model for studying the establishment of hepatocyte-specific GRNs. In this study, we utilized high-throughput single-cell RNA sequencing (scRNA-seq) to explore the cell-fate transition and the establishment of hepatocyte-specific GRNs involved in the two-step reprogramming process. Our findings revealed that the reprogramming process mimics the natural trajectory of liver development, exhibiting similar tran-scriptional waves of developmental genes. CD24 and DLK1 were identified as sur-face markers enriching two distinct hepatic progenitor populations respectively. Li-pid metabolism emerged as a key enhancer of hiHeps maturation. Furthermore, tran-scription factors HNF4A and HHEX were identified as pivotal gatekeepers directing cell fate decisions between hepatocytes and intestinal cells. Collectively, this study provides valuable insights into the establishment of hepatocyte-specific GRNs during hiHeps induction at single-cell resolution, facilitating more efficient production of functional hepatocytes for therapeutic applications.
Funding Source: This work was supported by the National Natural Science Foundation of China (32300675, 32400598), Sichuan Science and Technology Program (2022YFS0615, 2025ZNSFSC1019), Luzhou Science and Technology Program of China (2023SYF136)
