Aims Vascular smooth muscle cells (SMCs) are major precursors contributing to osteochondrogenesis and calcification in atherosclerosis. Runt-related transcription factor-2 (Runx2) has been found essential for SMC differentiation to an osteochondrogenic phenotype and subsequent calcification in vitro. A recent study using a conditional targeting allele that produced a truncated Runx2 protein in SMCs of ApoE?/? mice showed reduced vascular calcification, likely occurring via reduction of receptor activator of nuclear factor-?B ligand (RANKL), macrophage infiltration, and atherosclerotic lesion formation. Using an improved conditional Runx2 knockout mouse model, we have elucidated new roles for SMC-specific Runx2 in arterial intimal calcification (AIC) without effects on atherosclerotic lesion size.
Methods and results We used an improved targeting construct to generate LDLr?/? mice with floxed-Runx2 alleles (LDLr?/?:Runx2f/f) such that Cre-mediated recombination (LDLr?/?:Runx2?SM) does not produce functional truncated Runx2 protein, thereby avoiding off-target effects. We found that both LDLr?/?:Runx2f/f and LDLr?/?:Runx2?SM mice fed with a high fat diet developed atherosclerosis. SMC-specific Runx2 deletion did not significantly reduce atherosclerotic lesion size, macrophage number, or expression of RANKL, MCP-1, and CCR2. However, it significantly reduced AIC by 50%. Mechanistically, Sox9 and type II collagen were unaltered in vessels of LDLr?/?:Runx2?SM mice compared to LDLr?/?:Runx2f/f counterparts, while type X collagen, MMP13 and the osteoblastic marker osteocalcin were significantly reduced.
Conclusions SMC autonomous Runx2 is required for SMC differentiation towards osteoblast-like cells, SMC-derived chondrocyte maturation and AIC in atherosclerotic mice. These effects were independent of systemic lipid metabolism, RANKL expression, macrophage infiltration, and atheromatous lesion progression.