TY - JOUR T1 - Single cell trajectory analysis of human pluripotent stem cells differentiating towards lung and hepatocyte progenitors JF - bioRxiv DO - 10.1101/2021.02.23.432413 SP - 2021.02.23.432413 AU - Chaido Ori AU - Meshal Ansari AU - Ilias Angelidis AU - Fabian J. Theis AU - Herbert B. Schiller AU - Micha Drukker Y1 - 2021/01/01 UR - http://biorxiv.org/content/early/2021/02/23/2021.02.23.432413.abstract N2 - Understanding the development of human respiratory tissues is crucial for modeling and treating lung disorders. The molecular details for the specification of lung progenitors from human pluripotent stem cells (hPSCs) are unclear. Here, we use single cell RNA-sequencing with high temporal resolution along an optimized differentiation protocol to determine the transcriptional hierarchy of lung specification from human hPSCs and map out the underlying single cell trajectories. We show that Sonic hedgehog, TGF-β and Notch activation are required in an ISL1/NKX2-1 trajectory that gives rise to lung progenitors during the foregut endoderm stage. Induction of HHEX marks an alternative trajectory at the early definitive endoderm stage, which precedes the lung trajectory and generates a major hepatoblast population. Moreover, neither KDR+ nor mesendoderm progenitors are apparent intermediate states of lung and hepatic lineages. Our hierarchical multistep model predicts mechanisms leading to lung organogenesis, and creates a basis for studying early human lung development, as well as hPSC based disease and drug research.Competing Interest StatementThe authors have declared no competing interest. ER -