A novel agonist for the HGF receptor MET promotes differentiation of human pluripotent stem cells into hepatocyte‐like cells

Abstract

Hepatocyte growth factor (HGF) is the natural ligand of the MET receptor tyrosine kinase. This ligand-receptor couple is essential for the maturation process of hepatocytes. Previously, the rational design of a synthetic protein based on the assembly of two K1 domains from HGF led to the production of a potent and stable MET receptor agonist. In this study, we compared the effects of K1K1 with HGF during the differentiation of hepatocyte progenitors derived from human induced pluripotent stem cells (hiPSCs). In vitro, K1K1, in the range of 20 to 200 nM, successfully substituted for HGF and efficiently activated ERK downstream signaling. Analysis of the levels of hepatocyte markers showed typical liver mRNA and protein expression (HNF4α, albumin, alpha-fetoprotein, CYP3A4) and phenotypes. Although full maturation was not achieved, the results suggest that K1K1 is an attractive candidate MET agonist suitable for replacing complex and expensive HGF treatments to induce hepatic differentiation of hiPSCs. K E Y W O R D S differentiation, hepatocyte-like cells, human induced pluripotent stem cells (hiPSCs), MET agonist, peptide 1 | INTRODUCTION Hepatocytes are specialized cells that account for up to 80% of the cells in the liver. These cells possess unique characteristics (hexagonal morphology, having several nuclei, having a high number of mitochondria, and exhibiting a specific metabolism; Casotti & D'Antiga, 2019) that give them the ability to perform essential functions in the human body such as detoxification of hazardous molecules, conversion of ammonia to urea, regulation of glucose release, etc. Despite having the largest regenerative capability of all organs, the liver can fail to maintain functional hepatocytes because of acute or chronic diseases or severe poisoning (Gilgenkrantz & Collin de l'Hortet, 2018).

Down-Regulation of CYP3A4 by the KCa1.1 Inhibition Is Responsible for Overcoming Resistance to Doxorubicin in Cancer Spheroid Models

“…Plasma membrane depolarization by the inhibition of K Ca 1.1 may also be involved in the inactivation of the Akt signaling pathway through a decrease in the intracellular concentration of Ca 2+ . In addition to the Akt signaling pathway, the JNK and ERK signaling pathways are also important for the acquisition of DOX resistance and the transcriptional regulation of CYP3A4 in cancerous and non-cancerous cells [17,[27][28][29]. In the previous study, we showed the involvement of K Ca 3.1 in the regulation of JNK and ERK signaling pathways in macrophages [30].…”

“…No significant changes in the relative expression level of CEBPB transcripts were found by a single treatment with SC79 or NK252 in LNCaP, MG-63, and SW-1353 spheroid models (n = 4, p > 0.05) (Figures 10 and 11). In addition to the Akt-Nrf2 signaling pathway, the JNK/Nrf2 and ERK/Nrf2 pathways contribute to the acquisition of DOX resistance [17,27] and the regulation of CYP3A4 expression [28,29]. A recent study showed that the Ca 2+ -activated K + channel, K Ca 3.1, potentially regulates the JNK and ERK signaling pathways [30].…”