Biofabrication of synthetic human liver tissue with advanced programmable functions
Rodrigo M. Florentino,
Kazutoyo Morita,
Nils Haep,
Takashi Motomura,
Ricardo Diaz-Aragon,
Lanuza A.P. Faccioli,
Alexandra Collin de l’Hortet,
Zeliha Cetin,
Carla Frau,
Lawrence Vernetti,
Anna-Klara Amler,
Alexander Thomas,
Tobias Lam,
Lutz Kloke,
Kazuki Takeishi,
D. Lansing Taylor,
Ira J. Fox,
Alejandro Soto-Gutierrez
Affiliations
Rodrigo M. Florentino
Department of Pathology, University of Pittsburgh, Pittsburgh, PA, USA; Pittsburgh Liver Research Center, University of Pittsburgh, Pittsburgh, PA, USA
Kazutoyo Morita
Department of Pathology, University of Pittsburgh, Pittsburgh, PA, USA
Nils Haep
Department of Pathology, University of Pittsburgh, Pittsburgh, PA, USA
Takashi Motomura
Department of Pathology, University of Pittsburgh, Pittsburgh, PA, USA
Ricardo Diaz-Aragon
Department of Pathology, University of Pittsburgh, Pittsburgh, PA, USA
Lanuza A.P. Faccioli
Department of Pathology, University of Pittsburgh, Pittsburgh, PA, USA
Alexandra Collin de l’Hortet
Department of Pathology, University of Pittsburgh, Pittsburgh, PA, USA
Zeliha Cetin
Department of Pathology, University of Pittsburgh, Pittsburgh, PA, USA
Carla Frau
Department of Pathology, University of Pittsburgh, Pittsburgh, PA, USA
Lawrence Vernetti
Pittsburgh Liver Research Center, University of Pittsburgh, Pittsburgh, PA, USA; University of Pittsburgh Drug Discovery Institute, University of Pittsburgh, Pittsburgh, PA, USA; Department of Computational and Systems Biology, University of Pittsburgh, Pittsburgh, PA, USA
Department of Pathology, University of Pittsburgh, Pittsburgh, PA, USA; Department of Surgery and Science, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan
D. Lansing Taylor
Pittsburgh Liver Research Center, University of Pittsburgh, Pittsburgh, PA, USA; University of Pittsburgh Drug Discovery Institute, University of Pittsburgh, Pittsburgh, PA, USA; Department of Computational and Systems Biology, University of Pittsburgh, Pittsburgh, PA, USA
Ira J. Fox
Department of Computational and Systems Biology, University of Pittsburgh, Pittsburgh, PA, USA; Department of Surgery, Children’s Hospital of Pittsburgh of UPMC, University of Pittsburgh, Pittsburgh, PA, USA; McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA
Alejandro Soto-Gutierrez
Department of Pathology, University of Pittsburgh, Pittsburgh, PA, USA; Pittsburgh Liver Research Center, University of Pittsburgh, Pittsburgh, PA, USA; University of Pittsburgh Drug Discovery Institute, University of Pittsburgh, Pittsburgh, PA, USA; McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA; Corresponding author
Summary: Advances in cellular engineering, as well as gene, and cell therapy, may be used to produce human tissues with programmable genetically enhanced functions designed to model and/or treat specific diseases. Fabrication of synthetic human liver tissue with these programmable functions has not been described. By generating human iPSCs with target gene expression controlled by a guide RNA-directed CRISPR-Cas9 synergistic-activation-mediator, we produced synthetic human liver tissues with programmable functions. Such iPSCs were guide-RNA-treated to enhance expression of the clinically relevant CYP3A4 and UGT1A1 genes, and after hepatocyte-directed differentiation, cells demonstrated enhanced functions compared to those found in primary human hepatocytes. We then generated human liver tissue with these synthetic human iPSC-derived hepatocytes (iHeps) and other non-parenchymal cells demonstrating advanced programmable functions. Fabrication of synthetic human liver tissue with modifiable functional genetic programs may be a useful tool for drug discovery, investigating biology, and potentially creating bioengineered organs with specialized functions.
