Integrated single-cell analysis and mechanistic validation of LncRNA H19 in BMSC-mediated osteogenesis and potential...

Ostealgia in bone malignancies primarily results from bone loss, underscoring the importance of osteogenic induction as a vital therapeutic strategy. We investigated the role of Long noncoding RNA H19 (LncRNA H19) in regulating Bone Marrow Stromal Cell (BMSC) differentiation.

We first analyzed the single-cell transcriptomic landscape of the human bone marrow niche using the GSE253355 dataset, comprising 82,742 cells across 12 samples. Cell annotation and CytoTrace trajectory analysis were employed to define lineage potentials. Findings were validated in vitro by manipulating H19 levels in BMSCs and assessing mineralization and signaling pathways.

Single-cell analysis identified 35 distinct cell clusters (including Adipo-MSC, Fibro-MSC, Osteo-MSC, and Osteoblast). Among bone lineage populations, Osteo-MSCs and Osteoblasts exhibited the lowest differentiation potential scores, indicating terminal differentiation. Crucially, H19 was specifically upregulated in these mature osteogenic clusters. Consistent with this, H19 overexpression in vitro significantly enhanced mineralization, cell viability, and osteogenic markers (ALP, Runx2, Col1) by activating BMP2, NF-κB, and Wnt5a signaling. Conversely, H19 knockdown reversed these phenotypes.

LncRNA H19 acts as a lineage-specific driver of osteogenic differentiation via the BMP2/NF-κB/Wnt5a axis. Targeting H19 represents a promising approach to promote bone regeneration and potentially modulate pathways associated with cancer-associated ostealgia.