Hepatocyte-like cells from human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) are expected to be a useful source of cells drug discovery. Although we recently reported that hepatic commitment is promoted by transduction of SOX17 and HEX into human ESC- and iPSC-derived cells, these hepatocyte-like cells were not sufficiently mature for drug screening. To promote hepatic maturation, we utilized transduction of the hepatocyte nuclear factor 4α (HNF4α) gene, which is known as a master regulator of liver-specific gene expression. Adenovirus vector-mediated overexpression of HNF4α in hepatoblasts induced by SOX17 and HEX transduction led to upregulation of epithelial and mature hepatic markers such as cytochrome P450 (CYP) enzymes, and promoted hepatic maturation by activating the mesenchymal-to-epithelial transition (MET). Thus HNF4α might play an important role in the hepatic differentiation from human ESC-derived hepatoblasts by activating the MET. Furthermore, the hepatocyte like-cells could catalyze the toxication of several compounds. Our method would be a valuable tool for the efficient generation of functional hepatocytes derived from human ESCs and iPSCs, and the hepatocyte-like cells could be used for predicting drug toxicity.
Interindividual differences in hepatic metabolism, which are mainly due to genetic polymorphism in its gene, have a large influence on individual drug efficacy and adverse reaction. Hepatocyte-like cells (HLCs) differentiated from human induced pluripotent stem (iPS) cells have the potential to predict interindividual differences in drug metabolism capacity and drug response. However, it remains uncertain whether human iPSC-derived HLCs can reproduce the interindividual difference in hepatic metabolism and drug response. We found that cytochrome P450 (CYP) metabolism capacity and drug responsiveness of the primary human hepatocytes (PHH)-iPSHLCs were highly correlated with those of PHHs, suggesting that the PHH-iPS-HLCs retained donor-specific CYP metabolism capacity and drug responsiveness. We also demonstrated that the interindividual differences, which are due to the diversity of individual SNPs in the CYP gene, could also be reproduced in PHH-iPS-HLCs. We succeeded in establishing, to our knowledge, the first PHH-iPS-HLC panel that reflects the interindividual differences of hepatic drugmetabolizing capacity and drug responsiveness.human iPS cells | hepatocyte | CYP2D6 | personalized drug therapy | SNP D rug-induced liver injury (DILI) is a leading cause of the withdrawal of drugs from the market. Human induced pluripotent stem cell (iPSC)-derived hepatocyte-like cells (HLCs) are expected to be useful for the prediction of DILI in the early phase of drug development. Many groups, including our own, have reported that the human iPS-HLCs have the ability to metabolize drugs, and thus these cells could be used to detect the cytotoxicity of drugs that are known to cause DILI (1, 2). However, to accurately predict DILI, it will be necessary to establish a panel of human iPS-HLCs that better represents the genetic variation of the human population because there are large interindividual differences in the drug metabolism capacity and drug responsiveness of hepatocytes (3). However, it remains unclear whether the drug metabolism capacity and drug responsiveness of human iPS-HLCs could reflect those of donor parental primary human hepatocytes (PHHs). To address this issue, we generated the HLCs differentiated from human iPSCs which had been established from PHHs (PHH-iPS-HLCs). Then, we compared the drug metabolism capacity and drug responsiveness of PHH-iPS-HLCs with those of their parental PHHs, which are genetically identical to the PHH-iPS-HLCs.Interindividual differences of cytochrome P450 (CYP) metabolism capacity are closely related to genetic polymorphisms, especially single nucleotide polymorphisms (SNPs), in CYP genes (4). Among the various CYPs expressed in the liver, CYP2D6 is responsible for the metabolism of approximately a quarter of commercially used drugs and has the largest phenotypic variability, largely due to SNPs (5). It is known that certain alleles result in the poor metabolizer phenotype due to a decrease of CYP2D6 metabolism. Therefore, the appropriate dosage for drugs that are metabolized ...
Recent studies have indicated that there are many barriers to successful systemic gene delivery via cationic lipid vectors using the intravenous route. The purpose of this study was to investigate the effect of binding and interaction between erythrocytes, a major constituent of blood cells, and the complexes, in relation to the role of the helper lipid, on the in vivo gene delivery to the lung following intravenous injection. We used three types of cationic lipid vectors, DNA-DOTMA/Chol liposome complexes, DNA-DOTMA liposome complexes, and DNA-DOTMA/DOPE liposome complexes. Although the three types of vectors bind to murine blood cells in vivo and in vitro, DOTMA/Chol and DOTMA complexes with a higher in vivo transfection activity do not induce fusion between erythrocytes, whereas DOTMA/DOPE complexes, a less efficient vector in vivo,
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