Characterization of Genetically and Epigenetically Engineered Multicellular Systems and Their Application in Tissue Modeling and Hematopoiesis

Pluripotent stem cell (PSC)-derived organoids have emerged as novel multicellular models of human tissue development but display can be difficult to rationally engineer, display immature phenotypes, and have been adopted in limited applications. Here, using our preciously established human induced pluripotent stem cell (hiPSC)- derived multilineage human liver organoids we develop tools for epigenetic engineering liver organoids, demonstrate integrated analysis and engineering of gene regulatory networks (GRNs) in PSC-derived multilineage human liver organoids to direct maturation and vascular morphogenesis in vitro through transcription factor engineering, and establish a model of hematopoietic stem cell (HSC) colonization and expansion in the human fetal liver as a platform for expansion of therapeutic HSCs. Collectively, our approach provides an experimental framework to guide epigenetic and genetic engineering of PSC-derived organoids for tissue modeling and novel therapeutic applications.