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

doi:10.1016/j.isci.2020.101083

Efficient Generation and Transcriptomic Profiling of Human iPSC-Derived Pulmonary Neuroendocrine Cells

iScience. 2020 May 22;23(5):101083. doi: 10.1016/j.isci.2020.101083. Epub 2020 Apr 21.

Authors

Affiliations

¹ 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.
² Division of Hematology, Department of Medicine, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA.
³ 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.
⁴ Program in Cellular and Molecular Biology, University of Michigan Medical School, Ann Arbor, MI 48109, USA.
⁵ Lung and Regenerative Medicine Institutes, Department of Medicine, Cedars Sinai Medical Center, Los Angeles, CA 90048, USA.
⁶ Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO 63105, USA.
⁷ 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.
⁸ Department of Stem Cell Biology and Regenerative Medicine, Keck School of Medicine, HMR 712, University of Southern California, Los Angeles, CA 90033, USA. Electronic address: ichida@usc.edu.
⁹ 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. Electronic address: amy.firth@med.usc.edu.
¹⁰ 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. Electronic address: zborok@med.usc.edu.

Abstract

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.

Keywords: Cell Biology; Stem Cells Research; Transcriptomics.

Abstract

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