GSE138689
Single-cell transcriptomics of LepR-positive skeletal cells reveals heterogeneous stress-dependent stem and progenitor pools
Leptin receptor (LepR)-positive cells are key components of the bone marrow hematopoietic microenvironment, and highly enrich skeletal stem and progenitor cells that maintain homeostasis of the adult skeleton. However, the heterogeneity and lineage hierarchy within this population has been elusive. Using genetic lineage tracing and single-cell RNA sequencing, we found that Lepr-Cre labels most bone marrow stromal cells and osteogenic lineage cells in adult long bones. Integrated analysis of Lepr-Cre-traced cells under homeostatic and stress conditions revealed dynamic changes of the adipogenic, osteogenic, and periosteal lineages. Importantly, we discovered a Notch3+ bone marrow sub-population that is slow-cycling and closely associated with the vasculatures, as well as key transcriptional networks promoting osteo-chondrogenic differentiation. We also identified a Sca-1+ periosteal sub-population with high clonogenic activity but limited osteo-chondrogenic potential. Together, we mapped the transcriptomic landscape of adult LepR+ stem and progenitor cells and uncovered cellular and molecular mechanisms underlying their maintenance and lineage specification. Overall design: Single-cell transcriptomic profiling of Prrx1-Cre- and Lepr-Cre-traced stromal cells in steady state long bones from Prrx1-Cre;tdTomato;Col2.3-GFP and Lepr-Cre;tdTomato mice, respectively, as well as Lepr-Cre-traced stromal cells from Lepr-Cre;tdTomato mice under stress conditions such as aging, rosiglitazone feeding, irradiation and bone fracture. Bulk transcriptomic profiling of wild-type bone marrow stromal cells (BMSCs) cultured in 3D GelMA hydrogel alone, or 3D co-cultured with bone marrow endothelial cells (ECs).