Efficient Generation and Transcriptomic Profiling of Human iPSC-Derived Pulmonary Neuroendocrine Cells
Pooja Hor,
Vasu Punj,
Ben A. Calvert,
Alessandra Castaldi,
Alyssa J. Miller,
Gianni Carraro,
Barry R. Stripp,
Steven L. Brody,
Jason R. Spence,
Justin K. Ichida,
Amy L. Ryan (Firth),
Zea Borok
Affiliations
Pooja Hor
Hastings Center for Pulmonary Research and Division of Pulmonary, Critical Care and Sleep Medicine, Department of Medicine, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA; Department of Stem Cell Biology and Regenerative Medicine, Keck School of Medicine, HMR 712, University of Southern California, Los Angeles, CA 90033, USA
Vasu Punj
Division of Hematology, Department of Medicine, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA
Ben A. Calvert
Hastings Center for Pulmonary Research and Division of Pulmonary, Critical Care and Sleep Medicine, Department of Medicine, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA
Alessandra Castaldi
Hastings Center for Pulmonary Research and Division of Pulmonary, Critical Care and Sleep Medicine, Department of Medicine, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA
Alyssa J. Miller
Program in Cellular and Molecular Biology, University of Michigan Medical School, Ann Arbor, MI 48109, USA
Gianni Carraro
Lung and Regenerative Medicine Institutes, Department of Medicine, Cedars Sinai Medical Center, Los Angeles, CA 90048, USA
Barry R. Stripp
Lung and Regenerative Medicine Institutes, Department of Medicine, Cedars Sinai Medical Center, Los Angeles, CA 90048, USA
Steven L. Brody
Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO 63105, USA
Jason R. Spence
Program in Cellular and Molecular Biology, University of Michigan Medical School, Ann Arbor, MI 48109, USA; Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, MI 48109, USA; Department of Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor, MI 48109, USA; Department of Biomedical Engineering, University of Michigan College of Engineering, Ann Arbor, MI 48109, USA
Justin K. Ichida
Department of Stem Cell Biology and Regenerative Medicine, Keck School of Medicine, HMR 712, University of Southern California, Los Angeles, CA 90033, USA; Corresponding author
Amy L. Ryan (Firth)
Hastings Center for Pulmonary Research and Division of Pulmonary, Critical Care and Sleep Medicine, Department of Medicine, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA; Department of Stem Cell Biology and Regenerative Medicine, Keck School of Medicine, HMR 712, University of Southern California, Los Angeles, CA 90033, USA; Corresponding author
Zea Borok
Hastings Center for Pulmonary Research and Division of Pulmonary, Critical Care and Sleep Medicine, Department of Medicine, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA; Department of Biochemistry and Molecular Medicine, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA; Norris Comprehensive Center, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA; Corresponding author
Summary: Expansion of pulmonary neuroendocrine cells (PNECs) is a pathological feature of many human lung diseases. Human PNECs are inherently difficult to study due to their rarity (<1% of total lung cells) and a lack of established protocols for their isolation. We used induced pluripotent stem cells (iPSCs) to generate induced PNECs (iPNECs), which express core PNEC markers, including ROBO receptors, and secrete major neuropeptides, recapitulating known functions of primary PNECs. Furthermore, we demonstrate that differentiation efficiency is increased in the presence of an air-liquid interface and inhibition of Notch signaling. Single-cell RNA sequencing (scRNA-seq) revealed a PNEC-associated gene expression profile that is concordant between iPNECs and human fetal PNECs. In addition, pseudotime analysis of scRNA-seq results suggests a basal cell origin of human iPNECs. In conclusion, our model has the potential to provide an unlimited source of human iPNECs to explore PNEC pathophysiology associated with several lung diseases.
