Human pluripotent stem cell fate trajectories toward lung and hepatocyte progenitors

Chaido Ori; Meshal Ansari; Ilias Angelidis; Ruth Olmer; Ulrich Martin; Fabian J Theis; Herbert B Schiller; Micha Drukker

doi:10.1016/j.isci.2023.108205

Human pluripotent stem cell fate trajectories toward lung and hepatocyte progenitors

iScience. 2023 Oct 14;26(11):108205. doi: 10.1016/j.isci.2023.108205. eCollection 2023 Nov 17.

Authors

Chaido Ori¹, Meshal Ansari^{2

3}, Ilias Angelidis², Ruth Olmer^{4

5

6}, Ulrich Martin^{4

5

6}, Fabian J Theis^{3

7}, Herbert B Schiller^{2

8}, Micha Drukker^{1

9}

Affiliations

¹ Institute of Stem Cell Research, Helmholtz Munich, Neuherberg, Munich, Germany.
² Comprehensive Pneumology Center Munich (CPC-M), Institute of Lung Health and Immunity (LHI), Helmholtz Munich, Member of the German Center for Lung Research (DZL), Munich, Germany.
³ Department of Computational Health, Institute of Computational Biology, Helmholtz Munich, Munich, Germany.
⁴ Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery (HTTG), Hannover Medical School, 30625 Hannover, Germany.
⁵ Biomedical Research in Endstage and Obstructive Lung Disease (BREATH), Member of the German Center for Lung Research (DZL), Hannover Medical School, 30625 Hannover, Germany.
⁶ REBIRTH-Research Center for Translational and Regenerative Medicine, Hannover Medical School, 30625 Hannover, Germany.
⁷ TUM School of Life Sciences, Technical University of Munich, Munich, Germany.
⁸ Institute of Experimental Pneumology, LMU University Hospital, Ludwig-Maximilians University, Munich, Germany.
⁹ Division of Drug Discovery and Safety, Leiden Academic Centre for Drug Research (LACDR), Leiden University, Leiden, the Netherlands.

Abstract

In this study, we interrogate molecular mechanisms underlying the specification of lung progenitors from human pluripotent stem cells (hPSCs). We employ single-cell RNA-sequencing with high temporal precision, alongside an optimized differentiation protocol, to elucidate the transcriptional hierarchy of lung specification to chart the associated single-cell trajectories. Our findings indicate that Sonic hedgehog, TGF-β, and Notch activation are essential within an ISL1/NKX2-1 trajectory, leading to the emergence of lung progenitors during the foregut endoderm phase. Additionally, the induction of HHEX delineates an alternate trajectory at the early definitive endoderm stage, preceding the lung pathway and giving rise to a significant hepatoblast population. Intriguingly, neither KDR+ nor mesendoderm progenitors manifest as intermediate stages in the lung and hepatic lineage development. Our multistep model offers insights into lung organogenesis and provides a foundation for in-depth study of early human lung development and modeling using hPSCs.

Keywords: Cell biology; Developmental biology; Molecular biology.