Generation and molecular characterization of human pluripotent stem cell-derived pharyngeal foregut endoderm
Nicola A Kearns1, Macrina Lobo1, Ryan M J Genga1
1Program in Molecular Medicine, University of Massachusetts Chan Medical School, Worcester, MA, USA; Diabetes Center of Excellence, University of Massachusetts Chan Medical School, Worcester, MA, USA.
Insights
Researchers developed an efficient method to generate human pharyngeal foregut endoderm from pluripotent stem cells. This breakthrough aids in studying developmental disorders affecting the pharynx, thymus, parathyroid, and thyroid glands.
Area of Science:
- Developmental biology and stem cell research.
- Human organogenesis and developmental disorders.
Background:
- Studying human pharyngeal foregut endoderm is crucial for understanding syndromes affecting pharynx, thymus, parathyroid, and thyroid development.
- Limited tissue access and cellular models have hindered research in this area.
Purpose of the Study:
- To establish an efficient stepwise differentiation method for generating human pharyngeal foregut endoderm from pluripotent stem cells.
- To optimize differentiation protocols by determining signaling pathway requirements.
- To provide a resource for investigating developmental defects related to pharyngeal endoderm.
Main Methods:
- Stepwise differentiation of pluripotent stem cells.
- Dose and temporal optimization of signaling pathway engagement.
- Cellular and molecular characterization of differentiation products.
- Development of a computational classification tool (CellMatch).
- Transcriptomic classification using mouse scRNA-seq developmental roadmap.
- Integrated transcriptomic and chromatin analyses.
Main Results:
- An efficient method for generating human pharyngeal foregut endoderm was established.
- Optimized differentiation protocols were identified.
- Cellular maturation was confirmed via transcriptomic classification and a computational tool.
- Stage-specific gene regulatory networks were inferred through multiomic analyses.
Conclusions:
- The developed method provides a valuable resource for studying human pharyngeal foregut endoderm development.
- This approach facilitates research into the genetic basis of congenital disorders affecting the pharynx and its derivatives.
- The study offers insights into gene regulatory networks governing human foregut development.
Abstract:
Approaches to study human pharyngeal foregut endoderm-a developmental intermediate that is linked to various human syndromes involving pharynx development and organogenesis of tissues such as thymus, parathyroid, and thyroid-have been hampered by scarcity of tissue access and cellular models. We present an efficient stepwise differentiation method to generate human pharyngeal foregut endoderm from pluripotent stem cells. We determine dose and temporal requirements of signaling pathway engagement for optimized differentiation and characterize the differentiation products on cellular and integrated molecular level. We present a computational classification tool, "CellMatch," and transcriptomic classification of differentiation products on an integrated mouse scRNA-seq developmental roadmap confirms cellular maturation. Integrated transcriptomic and chromatin analyses infer differentiation stage-specific gene regulatory networks. Our work provides the method and integrated multiomic resource for the investigation of disease-relevant loci and gene regulatory networks and their role in developmental defects affecting the pharyngeal endoderm and its derivatives.
