Localized T3 production modifies the transcriptome and promotes the hepatocyte-like lineage in iPSC-derived hepatic organoids
Jorge Hidalgo-Álvarez, … , Tatiana L. Fonseca, Antonio C. Bianco
Jorge Hidalgo-Álvarez, … , Tatiana L. Fonseca, Antonio C. Bianco
Published October 19, 2023
Citation Information: JCI Insight. 2023;8(23):e173780. https://doi.org/10.1172/jci.insight.173780.
View: Text | PDF
Research Article Endocrinology Metabolism

Localized T3 production modifies the transcriptome and promotes the hepatocyte-like lineage in iPSC-derived hepatic organoids

Abstract

Thyroid hormone (TH) levels are low during development, and the deiodinases control TH signaling through tissue-specific activation or inactivation of TH. Here, we studied human induced pluripotent stem cell–derived (iPSC-derived) hepatic organoids and identified a robust induction of DIO2 expression (the deiodinase that activates T4 to T3) that occurs in hepatoblasts. The surge in DIO2-T3 (the deiodinase that activates thyroxine [T4] to triiodothyronine [T3]) persists until the hepatoblasts differentiate into hepatocyte- or cholangiocyte-like cells, neither of which expresses DIO2. Preventing the induction of the DIO2-T3 signaling modified the expression of key transcription factors, decreased the number of hepatocyte-like cells by ~60%, and increased the number of cholangiocyte-like cells by ~55% without affecting the growth or the size of the mature liver organoid. Physiological levels of T3 could not fully restore the transition from hepatoblasts to mature cells. This indicates that the timed surge in DIO2-T3 signaling critically determines the fate of developing human hepatoblasts and the transcriptome of the maturing hepatocytes, with physiological and clinical implications for how the liver handles energy substrates.

Authors

Jorge Hidalgo-Álvarez, Federico Salas-Lucia, Diana Vera Cruz, Tatiana L. Fonseca, Antonio C. Bianco

×

Figure 3

Formation of hepatic organoids in the presence or absence of TH.

Options: View larger image (or click on image) Download as PowerPoint
Formation of hepatic organoids in the presence or absence of TH. (A) Gra...
(A) Graphic representation of the development of hepatic organoids in the presence or absence of TH. The yellow arrow indicates the addition of TH (from day 5 to day 50). The final concentration of T4-HOs (red) and T3-HOs (blue) was 1 nM T4 (free T4 = ~15 pM) and 200 pM T3 (free T3 = ~10 pM); V-HOs were grown in the absence of T4 or T3 (black). C1–C6 are the indicated differentiation cocktails used (see methods). (B) Bright-field images (3 conditions) of hepatoblast at day 22 and hepatic organoids at day 29. Scale bar: 200 μm. (C) Relative mRNA levels of HNF4A (hepatocyte marker) and CEBPA from day 26 to day 32 (n = 4). (D) Albumin levels in the medium from day 35 to day 50 (n = 4). (E) Relative mRNA levels of KRT7 at day 46 and day 50 (n = 4, except T4-HOs, n = 3). (F) ALB/KRT7 mRNA ratio during from day 38 to day 46 (n = 4 except T4-HOs and T3-HOs at day 42, n = 2; T4-HOs at day 46, n = 3). (G and H) Apolipoprotein B (APOB) and Apolipoprotein A1 (APOA1) levels from the medium from day 35 to day 50, comparing V-HOs (black) versus T4-HOs (red). Ten organoids per well (n = 4). Two-tailed Student’s t test for comparing V-HOs versus T4-HOs, and 1-way ANOVA and Tukey test were used for multiple comparisons. Data are the mean of duplicates, represented as aligned scatter dot plots and their mean. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. The days of the differentiation are shown on the x axis (see legend Figure 1).