Authors: Jeremy J. Velazquez, Ryan LeGraw, Farzaneh Moghadam, Yuqi Tan, Jacquelyn Kilbourne, Joseph C. Maggiore, Joshua Hislop, Silvia Liu, Davy Cats, Susana M. Chuva de Sousa Lopes, Christopher Plaisier, Patrick Cahan, Samira Kiani, Mo R. Ebrahimkhani
Summary: Pluripotent stem cell (PSC)-derived organoids have emerged as novel multicellular models of human tissue development but display immature phenotypes, aberrant tissue fates, and a limited subset of cells. Here, we demonstrate that integrated analysis and engineering of gene regulatory networks (GRNs) in PSC-derived multilineage human liver organoids direct maturation and vascular morphogenesis in vitro. Overexpression of PROX1 and ATF5, combined with targeted CRISPR-based transcriptional activation of endogenous CYP3A4, reprograms tissue GRNs and improves native liver functions, such as FXR signaling, CYP3A4 enzymatic activity, and stromal cell reactivity. The engineered tissues possess superior liver identity when compared with other PSC-derived liver organoids and show the presence of hepatocyte, biliary, endothelial, and stellate-like cell populations in single-cell RNA-seq analysis. Finally, they show hepatic functions when studied in vivo. Collectively, our approach provides an experimental framework to direct organogenesis in vitro by systematically probing molecular pathways and transcriptional networks that promote tissue development.
Source: Cell Systems, published 12/07/2020