The nuclear receptor THRB facilitates differentiation of human PSCs into more mature hepatocytes

• DOI: 10.1016/j.stem.2022.03.015
• Publication Date: 2022-04-21T10:28:00Z
• Authors: Haiting Ma; Esmée de Zwaan; Yang Eric Guo; Paloma Cejas; Prathapan Thiru; Martijn van de Bunt; Jacob F. Jeppesen; Sudeepa Syamala; Alessandra Dall’A...; Brian J. Abraham; Dongdong Fu; Carrie Garrett-En...; Tong Ihn Lee; Henry W. Long; Linda G. Griffith; Richard A. Young; Rudolf Jaenisch
• Keywords: Mice; Hepatocytes; Animals; Humans; Receptors, Cytoplasmic and Nuclear; Cell Differentiation; Regulatory Sequences, Nucleic Acid; Polymorphism, Single Nucleotide; Chromatin

To understand the mechanisms regulating the in vitro maturation of hPSC-derived hepatocytes, we developed a 3D differentiation system and compared gene regulatory elements in human primary hepatocytes with those in hPSC-hepatocytes that were differentiated in 2D or 3D conditions by RNA-seq, ATAC-seq, and H3K27Ac ChIP-seq. Regulome comparisons showed a reduced enrichment of thyroid receptor THRB motifs in accessible chromatin and active enhancers without a reduced transcription of THRB. The addition of thyroid hormone T3 increased the binding of THRB to the CYP3A4 proximal enhancer, restored the super-enhancer status and gene expression of NFIC, and reduced the expression of AFP. The resultant hPSC-hepatocytes showed gene expression, epigenetic status, and super-enhancer landscape closer to primary hepatocytes and activated regulatory regions including non-coding SNPs associated with liver-related diseases. Transplanting the hPSC-hepatocytes resulted in the engraftment of human hepatocytes into the mouse liver without disrupting normal liver histology. This work implicates the environmental factor-nuclear receptor axis in regulating the maturation of hPSC-hepatocytes.