Developmental dynamics of two bipotent thymic epithelial progenitor types

T cell development in the thymus is essential for cellular immunity and depends on the organotypic thymic epithelial microenvironment. In comparison with other organs, the size and cellular composition of the thymus are unusually dynamic, as exemplified by rapid growth and high T cell output during early stages of development, followed by a gradual loss of functional thymic epithelial cells and diminished naive T cell production with age(1-10). Single-cell RNA sequencing (scRNA-seq) has uncovered an unexpected heterogeneity of cell types in the thymic epithelium of young and aged adult mice(11-18); however, the identities and developmental dynamics of putative pre- and postnatal epithelial progenitors have remained unresolved(1,12,16,17,19-27). Here we combine scRNA-seq and a new CRISPR-Cas9-based cellular barcoding system in mice to determine qualitative and quantitative changes in the thymic epithelium over time. This dual approach enabled us to identify two principal progenitor populations: an early bipotent progenitor type biased towards cortical epithelium and a postnatal bipotent progenitor population biased towards medullary epithelium. We further demonstrate that continuous autocrine provision of Fgf7 leads to sustained expansion of thymic microenvironments without exhausting the epithelial progenitor pools, suggesting a strategy to modulate the extent of thymopoietic activity.

Automated summary

T cell development in the thymus is dependent on the organotypic thymic epithelial microenvironment. Using a combination of scRNA-seq and a novel CRISPR-Cas9-based cellular barcoding system, this study identified two principal progenitor populations in the thymic epithelium and revealed qualitative and quantitative changes in the thymic epithelium over time.